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RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.


 
 

Reproduction, Fertility and Development publishes research and reviews on all aspects of vertebrate reproduction and developmental biology. More

Editor-in-Chief: Graeme Martin

 
 
 

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Published online 28 April 2016
Proteome of equine oviducal fluid: effects of ovulation and pregnancy 
Katrien Smits, Hilde Nelis, Katleen Van Steendam, Jan Govaere, Kim Roels, Cyrillus Ververs, Bart Leemans, Eline Wydooghe, Dieter Deforce and Ann Van Soom

Early development of the equine embryo occurs in the oviduct, and information on the composition of this oviducal environment is very valuable. We identified the proteins in the oviducal fluid of the horse, and proteins that are important for pregnancy were found in higher amounts when an equine embryo was present. These findings provide a solid basis for further studies on embryo–maternal interactions in the horse.

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Published online 26 April 2016
Sperm function during incubation with oestrus oviductal fluid differs in bulls with different fertility 
A. Kumaresan, A. Johannisson and A.-S. Bergqvist

Prediction of breeding bull fertility is of major economic value to the cattle-breeding industry. We proved that spermatozoa from high-fertile bulls showed a low degree of tyrosine phosphorylation and high degree of capacitation and acrosome reaction upon exposure to oviductal fluid. These findings may help in developing tools for assessing fertility of bulls, once validated in more animals.

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Published online 11 April 2016
Carnitine-mediated antioxidant enzyme activity and Bcl2 expression involves peroxisome proliferator-activated receptor-γ coactivator-1α in mouse testis 
Vikas Kumar Roy, Rachna Verma and Amitabh Krishna

Because carnitine treatment is used in many cases of male infertility, the aim of the present study was to evaluate the mechanism by which carnitine improves spermatogenesis in mice. In the study, carnitine treatment of mice stimulated peroxisome proliferator-activated receptor-γ coactivator-1α expression, which, in turn, regulated steroidogenesis by enhancing expression of Bcl2 and antioxidant enzymes in the testis.

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Published online 11 April 2016
Oestrogenic action of neonatal tamoxifen on the hypothalamus and reproductive system in female mice 
Rahmatollah Parandin, Morteza Behnam-Rassouli and Nasser Mahdavi-Shahri

Tamoxifen is widely used in the treatment of breast cancer and can interfere with the reproductive system. The present study showed that tamoxifen administration during the neonatal period has oestrogenic actions on oestrogen-sensitive nuclei in the brain controlling the reproductive axis in female mice. Therefore, tamoxifen may have adverse effects on brain differentiation and reproductive function.

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Published online 06 April 2016
Prion protein 2 (dublet) gene (PRND): role in ovine semen capacitation, cryopreservation and fertility 
L. M. Ferreira, M. Garcia-Herreros, A. Domingos, C. C. Marques, P. Mesquita, J. P. Barbas, M. C. Baptista, J. Pimenta, A. E. M. Horta, J. A. M. Prates and R. M. L. N. Pereira

The physiological functions of prion and prion-like proteins are overlapped by their pathological features, namely in the invariably fatal neurodegenerative diseases of both humans and animals, such as Kuru, Creutzfeldt-Jakob disease and new variant, bovine spongiform encephalopathy and sheep scrapie. The aim of the present study was to investigate the role of the prion-like Doppel (PRND) protein and different PRND genotypes in the sperm capacitation process, cryoresistance, and fertility potential. Both factors were crucial for sperm capacitation potential, cryoresistance, and embryo production. Thus, an important physiological function of Doppel protein and genotype in male fertility has been demonstrated.

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Published online 06 April 2016
Short- and long-term outcomes of the absence of protein during bovine blastocyst formation in vitro 
A. Murillo-Ríos, V. Maillo, M. Muñoz, A. Gutiérrez-Adán, S. Carrocera, D. Martín-González, A. Fernandez-Buznego and E. Gómez

Removal of protein from culture medium improves analytical reliability, although bovine embryo development in vitro could be affected. However, protein removal over a 24-h period in culture led to embryos with reduced miscarriage rates without affecting birthweight, gestation length or calf morphology. This finding represents direct access to culture medium as a substrate for non-invasive analysis, leading to potential biomarker discovery without compromising embryonic viability.

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Published online 06 April 2016
Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen 
J. M. Morrell, A. Lagerqvist, P. Humblot and A. Johannisson

Stallion spermatozoa lose viability and fertilising capability relatively quickly when cooled; robust spermatozoa can be selected by colloid centrifugation. In the present study, selected spermatozoa were shown to metabolise differently to controls, producing less hydrogen peroxide, which is known to damage spermatozoa. These results indicate that colloid centrifugation could be used to improve the quality of stallion semen for AI.

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Published online 09 March 2016
Morphometric developmental pattern of bovine handmade cloned concepti in late pregnancy 
R. P. C. Gerger, F. C. Zago, E. S. Ribeiro, S. Gaudencio Neto, L. T. Martins, L. H. Aguiar, V. H. V. Rodrigues, F. H. Furlan, I. Ortigari, R. D. Sainz, C. L. Ferrell, M. A. Miglino, C. E. Ambrósio, J. L. Rodrigues, R. Rossetto, F. Forell, L. R. Bertolini and M. Bertolini

Cloned concepti on Day 225 of gestation were characterised by lower pregnancy rates, high gestational losses, a distinctive biphasic growth pattern, with early growth restriction followed by accelerated growth and foetal weight on Day 225, and significant morphological changes in placental mass and shape when compared with in vivo-derived pregnancies. Conversely, Day-225 IVF-derived concepti were widely similar to controls.

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Published online 09 March 2016
Local effect of lysophosphatidic acid on prostaglandin production in the bovine oviduct 
Yuka Yoshimoto, Yoshihiko Kobayashi, Izabela Woclawek-Potocka, Emilia Sinderewicz, Yuki Yamamoto, Koji Kimura and Kiyoshi Okuda

Prostaglandins induce contraction and relaxation of oviductal smooth muscle, which are essential for transport of gametes and embryo. The present results demonstrate that lysophosphatidic acid significantly stimulated prostaglandin production by cultured isthmic stromal cells. It is suggested that locally produced lysophosphatidic acid contributes to oviductal motility in the isthmus by regulating prostaglandin production in cows.

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Published online 09 March 2016
Increased frequency of chromosome congression defects and aneuploidy in mouse oocytes cultured at lower temperature 
Jitka Danadova, Natalie Matijescukova, Anna Mac Gillavry Danylevska and Martin Anger

In vitro manipulation of oocytes is a standard procedure used in assisted reproduction. The aim of the present study was to analyse the effect of suboptimal culture temperatures on various aspects of meiotic division, including chromosomal congression and segregation defects. The results revealed that even a small deviation from optimal culture temperature has a marked effect on chromosome congression and aneuploidy.

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Published online 09 March 2016
Implementing the use of a biobank in the endangered black-footed ferret (Mustela nigripes) 
Rachel Santymire

The critically endangered black-footed ferret teeters on the edge of extinction with only approximately 600 left globally. With this genetically closed population, assisted reproductive technology has ensured every ferret passes his/her genes onto the next generation, and the use of a biobank has slowed the loss of gene diversity. With 30 years of experience, this recovery program serves as a model for other threatened wildlife.

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Published online 09 March 2016
Lower dietary n-6:n-3 ratio and high-dose vitamin E supplementation improve sperm morphology and oxidative stress in boars 
Qing Liu, Yuanfei Zhou, Runjia Duan, Hongkui Wei, Siwen Jiang and Jian Peng

Dietary PUFA and vitamin E are known to influence sperm morphology and oxidative stress. The effects of dietary n-6 : n-3 and vitamin E on the sperm morphology and oxidative stress in boars were investigated. An n-6 : n-3 ratio of 6 :1 and 400 mg kg–1 vitamin E have beneficial effects on sperm morphology by improving oxidative stress.

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Published online 07 March 2016
Expression of 17β-hydroxysteroid dehydrogenase and the effects of LH, FSH and prolactin on oestrone and 17β-oestradiol secretion in the endometrium of pigs during early pregnancy and the oestrous cycle 
B. Wojciechowicz, G. Kotwica, K. Zglejc, E. Waszkiewicz and A. Franczak

The uterus produces steroids and primary steroidogenic enzymes are expressed in the uterus of pregnant and cyclic pigs. In this study we examined 17β-hydroxysteroid dehydrogenase expression and the effects of LH, FSH and prolactin on oestrone and 17β-oestradiol release in vitro in endometrial explants. We show that LH, FSH and prolactin may regulate oestrogen release by the endometrium.

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Published online 07 March 2016
Immunolocalisation of aromatase regulators liver kinase B1, phosphorylated AMP-activated protein kinase and cAMP response element-binding protein-regulated transcription co-activators in the human testis 
Seungmin Ham, Kristy A. Brown, Evan R. Simpson and Sarah J. Meachem

The hormone oestrogen is essential for testis development and its synthesis is dependent on the enzyme aromatase. However, the molecular mechanism of aromatase regulation is still poorly understood. This study describes for the first time the cellular localisation of proteins important for aromatase regulation, which are well known in breast tissue, in the normal testis. This new finding may help develop new therapies for oestrogen-mediated infertility.

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Published online 07 March 2016
Lonidamine-ethyl ester-mediated remodelling of the Sertoli cell cytoskeleton induces phosphorylation of plakoglobin and promotes its interaction with ?-catenin at the blood–testis barrier 
Dolores D. Mruk, Michele Bonanomi and Bruno Silvestrini

We are in need of safe, effective and affordable male contraceptives. Indazole carboxylic acids such as lonidamine and adjudin disrupt adhesion between Sertoli and germ cells in the rat testis, resulting in infertility. One of the key findings of this study was that lonidamine-ethyl ester also displayed contraceptive effects. Additional studies are required to determine whether indazole carboxylic acids can enter the clinical pipeline

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Published online 02 March 2016
Serial transvaginal ultrasound-guided biopsy of the porcine corpus luteum in vivo 
S. Björkman, J. Yun, M. Niku, C. Oliviero, N. M. Soede and O. A. T. Peltoniemi

For the first time, a minimal invasive method that allows ovarian tissue collection from sows is described in detail. This method, a transvaginal ultrasound-guided biopsy, is quick, can be performed by one person, and has no major effects on the sow’s reproductive performance. It can be applied on the farm to study effects of housing or nutrition on luteal function.

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Published online 01 March 2016
Roles of intracellular ice formation, vitrification of cell water, and recrystallisation of intracellular ice on the survival of mouse embryos and oocytes 
Peter Mazur and Estefania Paredes

Mazur and collaborators have been studying the validity of initial views on vitrification in mouse oocytes and embryos. After decades of work they have established that survival is dependent on the dehydration prior to vitrification, as well as on warming rate. This new paradigm of vitrification may broaden the application of vitrification and ultra-fast warming to other cells. It remains to be seen how widely these findings will be applicable.

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Published online 29 February 2016
Insulin exposure during in vitro bovine oocyte maturation changes blastocyst gene expression and developmental potential 
Denise Laskowski, Ylva Sjunnesson, Patrice Humblot, Marc-André Sirard, Göran Andersson, Hans Gustafsson and Renée Båge

Metabolic imbalance impairs female fertility. We wanted to find out how metabolism affects bovine oocyte quality, and demonstrated that insulin exposure during in vitro oocyte maturation affects embryo development, morphology and gene expression. Even short-term exposure to a metabolic imbalance has lasting effects on embryo development, and this metabolic programming may have consequences later in life.

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Published online 29 February 2016
Greater sperm complexity in the Australasian old endemic rodents (Tribe: Hydromyini) is associated with increased levels of inter-male sperm competition 
Hanna J. McLennan, Stefan Lüpold, Pete Smissen, Kevin C. Rowe and William G. Breed

Most species of Australasian old endemic rodents have a highly complex sperm head morphology that contains an apical hook together with two ventral processes of variable length and orientation. This study tested whether this variation is a sexually selected trait using relative testes mass as a proxy for the intensity of inter-male sperm competition and controlling for phylogeny. The angle of these processes, but not length, was positively correlated with relative testes mass; its possible functional significance is briefly discussed.

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Published online 29 February 2016
Sperm freeze-drying and micro-insemination for biobanking and maintenance of genetic diversity in mammals 
Takehito Kaneko

Reproductive technologies are now indispensable tools for the efficient production of offspring in various mammals. This review covers the latest method of sperm freeze-drying and fertilization by intracytoplasmic sperm injection. Reproductive technologies provide a substantial advantage for biobanking and maintaining the genetic diversity of laboratory, domestic and wild animals.

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Published online 25 February 2016
Micromanipulation of equine blastocysts to allow vitrification 
Katrin Hinrichs and Young-Ho Choi

Cryopreservation of equine blastocysts is hindered by their large size and presence of an embryonic capsule. Collapse of the blastocoele via micromanipulation allows successful cryopreservation of embryos up to 600 microns diameter. Embryos may be shipped overnight to laboratories to perform the collapse and cryopreservation.

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Published online 24 February 2016
Implications of the Nagoya Protocol for genome resource banks composed of biomaterials from rare and endangered species 
Pierre Comizzoli and William V. Holt

The objective of the Nagoya Protocol is to prevent the unfair commercial exploitation of a country’s biodiversity. However, it has major effects on the ability of researchers from around the world to access non-commercial genetic materials as well. Herein we review the unforeseen implications of the Nagoya Protocol, as well as the possible solutions in relation to biobanking and animal conservation.

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Published online 24 February 2016
Fateful triad of reactive oxygen species, mitochondrial dysfunction and lipid accumulation is associated with expression outline of the AMP-activated protein kinase pathway in bovine blastocysts 
S. Prastowo, A. Amin, F. Rings, E. Held, D. Salilew Wondim, A. Gad, C. Neuhoff, E. Tholen, C. Looft, K. Schellander, D. Tesfaye and M. Hoelker

In vitro-derived bovine embryos frequently exhibit lipid accumulation, mitochondrial dysfunction and enhanced levels of reactive oxygen species. With the aim of unravelling the role of the AMP-activated protein kinase cytalytic subunit 1alpha (AMPKα), the present study identified AMPKα activity and distinct expression patterns of AMPKα-related genes to be correlated with that phenotype. These results prove the important role of AMPKα in embryonic energy metabolism.

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Published online 24 February 2016
Growth arrest specific 1 (Gas1) and glial cell line-derived neurotrophic factor receptor ?1 (Gfr?1), two mouse oocyte glycosylphosphatidylinositol-anchored proteins, are involved in fertilisation 
M. Agopiantz, L. Xandre-Rodriguez, B. Jin, G. Urbistondoy, C. Ialy-Radio, M. Chalbi, J.-P. Wolf, A. Ziyyat and B. Lefèvre

In addition to oocyte Cd9 tetraspanin, Juno, a glycosylphosphatidylinositol (GIP)-anchored protein, has recently been discovered to be essential to mammalian fertilisation, as a sperm receptor for Izumo1. The present study demonstrates the involvement of another two other oocyte GPI-anchored proteins, namely growth arrest specific 1 (Gas1) and glial cell line-derived neurotrophic factor receptor α1 (Gfrα1), in fertilisation. The results reinforce the implication of several molecules forming probably complexes.

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Published online 24 February 2016
Effect of spermatozoa motility hyperactivation factors and gamete coincubation duration on in vitro bovine embryo development using flow cytometrically sorted spermatozoa 
Luis B. Ferré, Yanina Bogliotti, James L. Chitwood, Cristóbal Fresno, Hugo H. Ortega, Michael E. Kjelland and Pablo J. Ross

This study evaluated the effects of sperm motility enhancers, media and different IVF times on embryo development. The sperm motility enhancers caffeine and theophylline improved cleavage and embryo development rates, synthetic oviducal fluid fertilisation medium outperformed M199 fertilisation medium and shortening the oocyte–sperm coincubation time (from 18 to 8 h) resulted in similar overall embryo performance rates. This study provides important information for optimising IVF protocols.

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Published online 15 February 2016
Detection of cross-sex chimerism in the common marmoset monkey (Callithrix jacchus) in interphase cells using fluorescence in situ hybridisation probes specific for the marmoset X and Y chromosomes 
E. Wedi, S. Müller, M. Neusser, P. H. Vogt, O. Y. Tkachenko, J. Zimmer, D. Smeets, H. W. Michelmann and P. L. Nayudu

Bone marrow chimerism is a unique and normal feature of Callithrichidae development that originates through shared placental circulation. A new X and Y chromosome-specific fluorescent in situ hybridisation protocol for Callithrix jacchus has been developed and validated to detect and investigate chimerism between male and female littermates. One of the strengths of this method is that it is useful in interphase cells, and allows cell morphology to remain intact to facilitate identification of cell type.

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Published online 15 February 2016
Obesity causes weight increases in prepubertal and pubertal male offspring and is related to changes in spermatogenesis and sperm production in rats 
Harish Navya and Hanumant Narasinhacharya Yajurvedi

Obesity, a global problem, adversely affects human health including reproduction. This study investigated whether exposure to an obesogenic environment during prenatal and postnatal periods affects the initiation and progress of spermatogenesis in rats. It is evident that an obese condition decreases counts of spermatids and spermatozoa and serum levels of testosterone and increases the proportion of abnormal spermatozoa, which might reduce reproductive potential.

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Published online 15 February 2016
Dietary-induced gestational iron deficiency inhibits postnatal tissue iron delivery and postpones the cessation of active nephrogenesis in rats 
Mary Y. Sun, Joseph C. Woolley, Sharon E. Blohowiak, Zachary R. Smith, Ashajyothi M. Siddappa, Ronald R. Magness and Pamela J. Kling

Gestational iron deficiency can cause developmental programming of adult hypertension, but the critical developmental window during nephrogenesis has not been studied. The present study examined renal development during perinatal iron deficiency in rats and found altered timing of kidney maturation and histological renal adaptations that may contribute to dysfunctional glomeruli and renal hyperfiltration, which themselves contribute to hypertension.

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Published online 11 February 2016
MicroRNA indicators of follicular steroidogenesis 
F. X. Donadeu, S. D. Sontakke and J. Ioannidis

Finding novel biomarkers of ovarian follicle function would be of benefit. This study investigated the follicle biomarker potential of microRNAs (miRNAs) and found that levels of miR-873 and miR-202 can be used as indicators of steroidogenic capacity in cattle. These miRNAs, in addition or alternatively to commonly used steroid level measurements, could be used to accurately classify follicles based on steroidogenic capacity.

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Published online 05 February 2016
Preantral follicle density in ovarian biopsy fragments and effects of mare age 
K. A. Alves, B. G. Alves, G. D. A. Gastal, K. T. Haag, M. O. Gastal, J. R. Figueiredo, M. L. Gambarini and E. L. Gastal

Information about follicle density is critical to determine the quantity of fragments during biopsy procedures in women and domestic animals. This study evaluated the effects of age and determined the minimum number of ovarian fragments and histological sections required to estimate equine follicle density in ovarian biopsy fragments. The results show that follicle density is negatively influenced by increasing age and, independent of follicle heterogeneity, three to four ovarian fragments combined with 65 histological sections are required to accurately determine follicle density in equine ovarian tissue.

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Published online 05 February 2016
Glucocorticoid-induced changes in glucocorticoid receptor mRNA and protein expression in the human placenta as a potential factor for altering fetal growth and development 
Svetlana Bivol, Suzzanne J. Owen and Roselyn B. Rose'Meyer

Glucocorticoids (GCs) sustain life and the development of human embryo; however, fetoplacental exposure to high maternal GC levels may lead to placental and fetal abnormalities, with the adverse consequences of GC excess manifested later in life. The mechanisms through which GC excess alters placental and fetal development are yet to be identified; however, changes in placental GR levels and subcellular localization and ratios may modify placental function, in utero growth of the fetus and lifelong health.

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Published online 02 February 2016
Age-associated deterioration in follicular fluid induces a decline in bovine oocyte quality 
Shun Takeo, Koji Kimura, Koumei Shirasuna, Takehito Kuwayama and Hisataka Iwata

The average age for a primipara has been increased and maternal age affects the quality of oocytes. The present study addressed whether age-associated deterioration of oocytes stems from the oocyte itself or the surrounding environment and found that follicular fluid plays a major role. On the basis of the results, given optimum conditions, oocytes derived from aged females could give rise to high-quality oocytes.

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Published online 14 January 2016
Effects of prepartum oilseed supplements on subclinical endometritis, pro- and anti-inflammatory cytokine transcripts in endometrial cells and postpartum ovarian function in dairy cows 
Reza Salehi, Marcos G. Colazo, Mohanathas Gobikrushanth, Urmila Basu and Divakar J. Ambrose

Dietary fats are known to influence immune cell function and fertility. The effects of pre-calving diets containing oilseeds (sunflower or canola vs. no oilseed) on uterine inflammation and ovarian function in dairy cows were investigated. Adding sunflower seed to pre-calving diets positively influenced ovarian function without affecting uterine inflammation, suggesting a role for omega-6 fatty acids in nutritional management of reproduction.

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Published online 12 January 2016
Genome-wide analysis of sperm DNA methylation from monozygotic twin bulls 
Habib A. Shojaei Saadi, Éric Fournier, Christian Vigneault, Patrick Blondin, Janice Bailey and Claude Robert

Monozygotic (MZ) twins are of great interest to elucidate the contributions of pre- and postnatal environmental factors on the expression of complex traits and diseases. In this study we investigated variations in sperm DNA methylation between MZ twin bulls to understand the observed differences in offspring productivity of MZ twin bulls despite their identical genetic backgrounds. Variation in sperm DNA methylation between MZ twin bulls was observed and may contribute to incongruous diverging performances of daughters sired by these bulls.

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Published online 11 January 2016
Mammalian target of rapamycin/eukaryotic initiation factor 4F pathway regulates follicle growth and development of theca cells in mice 
Chao Zhang, Xiao-Ran Liu, Yong-Chun Cao, Jin-Ling Tian, Di Zhen, Xiao-Fei Luo, Xin-Mei Wang, Jian-Hui Tian and Jian-Ming Gao

Follicular growth is mediated by gonadotropic hormones, growth factors and signalling pathways, such as the mammalian target of rapamycin (mTOR) signalling pathway. However, the molecular mechanisms underlying the development of theca cells have rarely been studied. Herein we provide unbiased, detailed and convincing evidence (microarray, in vitro and in vivo studies) to demonstrate that mTOR/eukaryotic initiation factor 4F signalling plays an important role in follicular growth, particularly the development of thecal cells.

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Published online 07 January 2016
Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development 
Leticia Torres, E. Hu and Terrence R. Tiersch

The lack of comprehensive quality assurance and quality control programs has impeded the use of cryopreservation in aquatic species, delaying the establishment of repositories and commercial-scale applications. In this review we focus on the opportunities for standardisation throughout the cryopreservation process and introduce key factors that would assist the transition to commercial-scale application. The establishment of germplasm repositories is essential for conservation efforts, stock enhancement for wild fisheries and genetic improvement for aquaculture.

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Published online 06 January 2016
LIM kinase activity is required for microtubule organising centre positioning in mouse oocyte meiosis 
Xin Li, Yubo Zhu, Yan Cao, Qian Wang, Juan Du, Jianhui Tian, Yuanjing Liang and Wei Ma

In female meiosis, accurate chromosome separation is driven by the spindle apparatus, which is organised under the regulation of unique microtubule organising centres (MTOCs). LIM kinase 1 activity is required for the establishment and maintenance of MTOC bipolarity in mouse oocytes during meiotic division. The results of the present study provide an insight into the molecular mechanism governing bipolar spindle formation in oocytes during meiotic maturation.

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Published online 04 January 2016
Active 3ʹ–5ʹ cyclic nucleotide phosphodiesterases are present in detergent-resistant membranes of mural granulosa cells 
Annick Bergeron, Christine Guillemette, Marc-André Sirard and François J. Richard

Lipids rafts are specialised membrane microdomains involved in cell signalling and phosphodiesterases are key enzymes involved in cyclic AMP signalling in the ovary. We found that in mural granulosa cell membranes, the cyclic AMP–phosphodiesterase activity is mainly contributed by the rafts. These results suggest that in mural granulosa cells, lipid rafts exist as a cell-signalling platform.

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Published online 18 December 2015
Biobanking genetic resources: challenges and implementation at the USDA National Animal Germplasm Program 
P. H. Purdy, C. S. Wilson, S. F. Spiller and H. D. Blackburn

Collecting genetic resources for preserving breeds or species, or for genetic analyses, is challenging for all agricultural species but especially difficult for minor breeds because of low numbers of animals, a lack of breed associations, and minimal information that can be used for categorisation of the animals. Since 1999 the USDA National Animal Germplasm Program has worked to overcome these obstacles and create a germplasm repository for all agricultural species in the USA. This manuscript describes these activities and illustrates them via a case study that describes how our efforts collecting Navajo-Churro sheep have created a secure backup of germplasm and how we specifically overcome these issues as they relate to rare and minor breeds of agricultural species.

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Published online 18 December 2015
Body mass index and human sperm quality: neither one extreme nor the other 
E. M. Luque, A. Tissera, M. P. Gaggino, R. I. Molina, A. Mangeaud, L. M. Vincenti, F. Beltramone, J. Sad Larcher, D. Estofán, M. Fiol de Cuneo and A. C. Martini

To determine whether body weight has an effect on semen quality, the present study explored the possible association between body weight (in terms of body mass index) and sperm function. The results indicate that both morbid obesity and being underweight have a negative effect on sperm quality, probably as a result of changes to epididymal function. These findings highlight the importance of an adequate body weight as the best option for fertility.

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Published online 18 December 2015
Elimination of methylation marks at lysines 4 and 9 of histone 3 (H3K4 and H3K9) of spermatozoa alters offspring phenotype 
Serafín Pérez-Cerezales, Priscila Ramos-Ibeas, Angela Lopez-Cardona, Eva Pericuesta, Raúl Fernandez-Gonzalez, Belen Pintado and Alfonso Gutiérrez-Adán

The function of epigenetic marks present in retained histones of mature spermatozoa is largely unknown. By reducing the methylation level at lysines 4 and 9 of histone 3 of mature spermatozoa we aimed to shed light on their significance during development. We discovered that these marks are important for normal embryo development and their reduction leads to abortions, premature death and morphological abnormalities in the offspring.

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Published online 18 December 2015
Effect of inbreeding depression on bull sperm quality and field fertility 
Jesús Dorado, Rosa Morales Cid, Antonio Molina, Manuel Hidalgo, Julia Ariza, Miguel Moreno-Millán and Sebastián Demyda-Peyrás

We studied for the first time the effect of high inbreeding levels in cattle sperm by automatised and objective methods. An extensive genetic analysis using an animal model showed that inbred bulls have a reduced field fertility and an increased calving interval compared with non-inbred bulls. Sperm from high inbred animals showed an increased ‘hyper-active like’ movement that could explain their decreased fertility.

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Published online 18 December 2015
Aquaporins in boar spermatozoa. Part II: detection and localisation of aquaglyceroporin 3 
Noelia Prieto-Martínez, Roser Morató, Ingrid Vilagran, Joan E. Rodríguez-Gil, Sergi Bonet and Marc Yeste

The aquaporin family plays a fundamental role in water and solute transport across biological membranes. The present work has studied, for the first time, the presence and localisation of aquaglyceroporin 3 in boar spermatozoa. Although two different localisation patterns for this protein were identified, neither AQP3 content nor its localisation were found to be correlated with sperm quality parameters.

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Published online 14 December 2015
In vitro maturation affects chromosome segregation, spindle morphology and acetylation of lysine 16 on histone H4 in horse oocytes 
Federica Franciosi, Ghylene Goudet, Irene Tessaro, Pascal Papillier, Rozenn Dalbies-Tran, Fabrice Reigner, Stefan Deleuze, Cecile Douet, Ileana Miclea, Valentina Lodde and Alberto M. Luciano

Mammalian eggs are often affected by aneuploidy, leading to failure in embryo implantation, miscarriages and genetic diseases. In horse oocytes the onset of aneuploidy is correlated with the disruption of an epigenetic mark that occurs without changes in gene expression. A deeper knowledge of the molecular basis of aneuploidy will allow the development of safer treatments to obtain healthy eggs.

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    | Supplementary Material (5.9 MB)
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Published online 30 November 2015
Soluble Delta-like ligand 1 alters human endometrial epithelial cell adhesive capacity 
Michelle Van Sinderen, Jennifer Oyanedel, Ellen Menkhorst, Carly Cuman, Katarzyna Rainczuk, Amy Winship, Lois Salamonsen, Tracey Edgell and Evdokia Dimitriadis

Infertility affects 1 in 10 couples or millions of people worldwide and while IVF is an important intervention, 75% of all embryos transferred into the uterus do not establish a pregnancy. We investigated how the uterus of infertile women may prevent pregnancy and discovered a protein released by uterine tissue that potentially blocks pregnancy. Targeting this protein may be useful as a new treatment option for infertile women.

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Published online 24 November 2015
Cumulus cell-conditioned medium supports embryonic stem cell differentiation to germ cell-like cells 
Syed Mohmad Shah, Neha Saini, Syma Ashraf, Manoj Kumar Singh, Radhey Sham Manik, Suresh Kumar Singla, Prabhat Palta and Manmohan Singh Chauhan

The effect of cumulus cell-conditioned medium (CCM) on the differentiation of buffalo embryonic stem cells to germ cells were explored. A concentration of 20%–40% conditioned medium induced the highest expression of primordial germ cell, meiotic, spermatocyte and oocyte markers and led to the formation of oocyte-like structures. Further development of the cumulus conditioned medium induced differentiation strategy would be useful for understanding mammalian gametogenesis and transgenic animal production.

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    | Supplementary Material (334 KB)
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Published online 13 November 2015
Is the association between insulin resistance and diabetogenic haematopoietically expressed homeobox (HHEX) polymorphism (rs1111875) affected by polycystic ovary syndrome status? 
F. Ramezani Tehrani, M. Zarkesh, M. Tohidi, F. Azizi and A. Zadeh-Vakili

Insulin resistance (IR) is an underlying factor, common to both polycystic ovary syndrome (PCOS) and type 2 diabetes. Genetic factors have also been reported to play a role in the pathogenesis of these two conditions, indicting a common genetic aetiology. In this study we aim to determine whether the genetic association between IR and diabetogenic SNPs is affected by PCOS status in Iranian women and found that the association may be influenced by the condition.

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Published online 29 October 2015
Follicular fluid and serum metabolites in Holstein cows are predictive of genetic merit for fertility 
S. G. Moore, A. O'Gorman, L. Brennan, T. Fair and S. T. Butler

This study examined the metabolite profile of serum and follicular fluid on Day 7 of the oestrous cycle in Holstein cows with good or poor genetic merit for fertility. Differences between the two genotypes in the abundance of fatty acids and amino acids were highly predictive of fertility. The results demonstrate that metabolite profiles in cattle respond to genetic selection for fertility traits, and specific metabolites may be useful biomarkers for fertility

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Published online 22 October 2015
Suppression of Sertoli cell tumour development during the first wave of spermatogenesis in inhibin α-deficient mice 
Jenna T. Haverfield, Peter G. Stanton, Kate L. Loveland, Heba Zahid, Peter K. Nicholls, Justine S. Olcorn, Yogeshwar Makanji, Catherine M. Itman, Evan R. Simpson and Sarah J. Meachem

Testicular cancer is the most common malignancy among young men; however, the mechanisms driving tumour growth are unclear. Herein we show that Sertoli cell tumours in the adult mouse testis can be profoundly suppressed by short-term inhibition of the pituitary hormone FSH in early postnatal life. These results show that FSH protects against Sertoli cell tumour growth during discrete windows of testicular development.

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    | Supplementary Material (104 KB)
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Published online 14 October 2015
Retinoic acid triggers c-kit gene expression in spermatogonial stem cells through an enhanceosome constituted between transcription factor binding sites for retinoic acid response element (RARE), spleen focus forming virus proviral integration oncogene (SPFI1) (PU.1) and E26 transformation-specific (ETS) 
Swanand Koli, Ayan Mukherjee and Kudumula Venkata Rami Reddy

Regulation of c-kit during spermatogenesis remains largely unknown. Retinoic acid (RA) regulates c-kit expression through discrete promoter regions. An enhanceosome constituted between transcription factor binding sites RARE, PU.1 and ETS in response to RA play an imperative role in c-kit activation. RA-induced c-kit regulation may provide insights in the understanding of nuances between self renewal and differentiation of spermatogonial cells.

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    | Supplementary Material (1.1 MB)
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Published online 14 October 2015
Sperm DNA fragmentation in cryopreserved samples from subjects with different cancers 
Lara Tamburrino, Marta Cambi, Sara Marchiani, Ilaria Manigrasso, Selene Degl'Innocenti, Gianni Forti, Mario Maggi, Elisabetta Baldi and Monica Muratori

Is sperm DNA fragmentation (SDF) involved in the lower fertilising ability of cryopreserved spermatozoa from cancer patients? In this study we evaluated SDF in two different cytometric sperm populations (PIbrighter and PIdimmer). In cryopreserved spermatozoa from cancer patients, PIbrighter SDF, which is more relevant to reproduction, was positively related to sperm motility and that the proportion of these cells was higher in samples from cancer versus non-cancer (oligozoospermic or affected by autoimmune pathologies) patients. The results of the study indicate that thawed motile spermatozoa of cancer patients may carry DNA damage.

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Published online 14 October 2015
The assessment of sperm DNA fragmentation in the saltwater crocodile (Crocodylus porosus) 
S. D. Johnston, C. López-Fernández, F. Arroyo, J. L. Fernández and J. Gosálvez

It is now possible to reliably collect crocodile semen for fertility evaluation and AI. Herein we describe a validated method for assessing the quality of crocodile sperm DNA using the sperm chromatin dispersion test. This technique, a first for any reptile, will now be used for refining improved methods of crocodile sperm preservation.

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Published online 14 October 2015
Effect of bovine oviductal fluid on development and quality of bovine embryos produced in vitro 
Ricaurte Lopera-Vasquez, Meriem Hamdi, Veronica Maillo, Valeriano Lloreda, Pilar Coy, Alfonso Gutierrez-Adan, Pablo Bermejo-Alvarez and Dimitrios Rizos

The post-fertilisation culture environment in vitro determines embryo quality, and different strategies have been developed to improve culture conditions. The use of bovine oviductal fluid supplementation during in vitro culture had a positive effect on embryo development and embryo quality. Thus, by mimicking in vivo conditions using animal models, we could improve assisted reproductive technologies applied to both domestic species and humans.

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Published online 14 October 2015
Peri-ovulatory endocrine regulation of the prostanoid pathways in the bovine uterus at early dioestrus 
Milena Lopes Oliveira, Fabio Luiz D'Alexandri, Guilherme Pugliesi, Veerle Van Hoeck, Fernando Silveira Mesquita, Claudia M. B. Membrive, João Alberto Negrão, Craig E. Wheelock and Mario Binelli

In cattle, sex steroids modulate pre-implantation uterine function, embryo receptivity and consequently, fertility. Prostaglandins (PGs) are synthesised in the endometrium and influence conceptus growth and corpus luteum lifespan. Here, we examined the biosynthetic pathway of PGs and determined that distinctly different pro-oestrus concentrations of oestradiol and dioestrus concentrations of progesterone did not affect endometrial PG production during the first week after oestrus. The influence of PGs in fertility occurs at later time points of gestation.

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    | Supplementary Material (31 KB)
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Published online 14 October 2015
Cytokine gene expression at the maternal–fetal interface after somatic cell nuclear transfer pregnancies in small ruminants 
Heloisa M. Rutigliano, Amanda Wilhelm, Justin Hall, Bi Shi, Qinggang Meng, Rusty Stott, Thomas D. Bunch, Kenneth L. White, Christopher J. Davies and Irina A. Polejaeva

The reasons why somatic cell nuclear transfer (SCNT) pregnancies are at increased risk of pregnancy loss are poorly understood. This study investigated pregnancy losses and gene expression of inflammatory mediators in sheep and goat SCNT pregnancies. Sheep SCNT pregnancies had an increased incidence of pregnancy loss and increased expression of inflammatory cytokines in the placenta, which indicates the presence of an unfavourable environment for fetal development.

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Published online 09 October 2015
Caudal vena cava progesterone and LH release patterns on Day 14 of gestation in primiparous sows 
L. L. Hoving, S. M. Haen, B. F. A. Laurenssen, O. A. T. Peltoniemi, B. Kemp and N. M. Soede

To date, not much is known about the interaction of progesterone and LH, nor progesterone’s metabolism during early pregnancy in the sow. This study of the release pattern and concentration of progesterone in the caudal vena cava, following LH pulses revealed their temporary relation and confirmed postprandial decrease of progesterone. Already on day 14 of an individual sow’s second pregnancy, the function of the CL seems to be affected by LH.

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Published online 08 October 2015
Characterisation and cryopreservation of the ovarian preantral follicle population from Spix 
Érica C. G. Praxedes, Gabriela L. Lima, Andréia M. Silva, Carlos A. C. Apolinário, José A. B. Bezerra, Ana L. P. Souza, Moacir F. Oliveira, Ana P. R. Rodrigues and Alexandre R. Silva

Spix’s yellow-toothed cavies (Galea spixii) are hystricomorph rodents. The ovarian preantral follicle (PF) population of G. spixii was characterised in the present study. It was estimated that the PF population per ovarian pair was 416.0 ± 342.8 (mean ± s.e.m.). Moreover, after a solid surface vitrification process using dimethyl sulfoxide as the cryoprotectant, 69.5% of PFs were morphologically normal.

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Published online 08 October 2015
Thyroid function and autoimmunity during ovarian stimulation for intracytoplasmic sperm injection 
Gesthimani Mintziori, Dimitrios G. Goulis, Efstratios M. Kolibianakis, Aristidis Slavakis, Julia Bosdou, Grigorios Grimbizis and Basil C. Tarlatzis

The role of thyroid hormones in implantation and early embryo development has been highlighted recently because of the increased use of assisted reproduction technology (ART). Close examination of thyroid function in women undergoing ovarian stimulation shows that thyrotropin-stimulating hormone (TSH) concentrations could increase during the procedure, particularly in women who become pregnant. If this is true, special care, such as universal baseline screening, has to be considered for women undergoing ART.

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Published online 05 October 2015
Modulation of embryonic development due to mating with immunised males 
Ludmila A. Gerlinskaya, Svetlana O. Maslennikova, Margaret V. Anisimova, Nataly A. Feofanova, Evgenii L. Zavjalov, Galina V. Kontsevaya, Yuri M. Moshkin and Mikhail P. Moshkin

Can paternal experiences be transmitted to offspring? Here we show that mating of immunised male mice with unimmunised females significantly altered gestation conditions and in utero embryo development resulting in increased embryonic bodyweight. These results emphasise that, as active environmental explorers, males confer their experiences to their progeny, suggesting an important mechanism of population-wise adaptation to infectious load.

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Published online 01 October 2015
The morphological and functional development of the stria vascularis in miniature pigs 
Weiwei Guo, Haijin Yi, Zhang Yan, Lili Ren, Lei Chen, Li Dong Zhao, Yu Ning, David Z. Z. He and Shi-Ming Yang

Our study evaluated how the stria vascularis and organ of Corti develop in miniature pigs. The stria vascularis develops during the embryonic period, concurrent with maturation of the organ of Corti in miniature pigs. The magnitude of endocochlear potential reached a steady state when the stria vascularis was morphologically adult-like at E98. These findings provide a morphological and functional basis for future animal studies of various inner-ear diseases.

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Published online 28 September 2015
Placenta-derived extracellular vesicles: their cargo and possible functions 
Mary Familari, Tina Cronqvist, Zahra Masoumi and Stefan R. Hansson

Given the great potential of extracellular vesicles as biomarkers for detecting early-stage disease – in particular pre-eclampsia – here we have reviewed the characteristics of placenta-shed vesicles and conducted a meta analysis of their protein content, which has revealed significant variability. Achieving the goal of biomarkers for pregnancy disorders will depend on rigorous standards for isolation and detection of placenta-derived vesicles.

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    | Supplementary Material (75 KB)
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Published online 25 September 2015
Inducing ovulation with oestradiol cypionate allows flexibility in the timing of insemination and removes the need for gonadotrophin-releasing hormone in timed AI protocols for dairy cows 
R. M. Ferreira, H. Ayres, L. U. Gimenes, F. P. Torres, F. A. Lima, M. B. Veras, T. G. Guida, R. V. Sala and P. S. Baruselli

The induction of synchronous ovulation and determination of the time of insemination are critical steps for the success of timed AI (TAI) in dairy cattle. The addition of gonadotrophin-releasing hormone (GnRH) had no beneficial effect on tightening the time to ovulation when oestradiol cypionate was used, and similar pregnancy rates were achieved when AI was performed 48 or 54 h after device removal. Thus, inducing ovulation with oestradiol cypionate removes the need for GnRH and can be a cost-effective and flexible option for TAI in dairy herds.

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Published online 22 September 2015
Cytoplasmic membrane activities during first cleavage of zona-free porcine embryos: description and consequences 
Rong Li, Ying Liu, Hanne Skovsgaard Pedersen and Henrik Callesen

Eggs are used in mammals for experiments and fertility treatment, but new ways to determine their quality are still needed. Using the pig as a model, eggs had their surrounding shell removed to visualise finer details in cell membranes during cleavage, showing new patterns reflecting their normality or otherwise. These observations supplement traditional egg evaluations, but require validation in other types of embryos from other mammals.

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    | Supplementary Material (2.6 MB)
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Published online 21 September 2015
Group management influences reproductive function of the male cheetah (Acinonyx jubatus) 
Diana C. Koester, Elizabeth W. Freeman, David E. Wildt, Kimberly A. Terrell, Ashley D. Franklin, Karen Meeks and Adrienne E. Crosier

The cheetah population under human care is notoriously difficult to breed and scientists need to gather as much biological data as possible to overcome issues ranging from behavioural compatibility to infertility. This work found that male cheetahs managed in small, closely bonded groups experience increased testicular function, including higher sperm quality, compared to males housed singly. These findings support existing management practices to improve breeding efforts to build assurance populations of cheetahs.

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Published online 21 September 2015
Nutrient restriction in early ovine pregnancy stimulates C-type natriuretic peptide production 
Sengodi Madhavan, Timothy C. R. Prickett, Eric A. Espiner and Graham K. Barrell

C-type natriuretic peptide (CNP) may be required for support of the developing conceptus during pregnancy. We explored this by placing pregnant ewes on a restricted diet and found that their blood levels of CNP were elevated in comparison with ewes on a normal level of nutrition. Elevation of CNP levels may be an adaptation to enhance placental function when fetal development is impaired.

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Published online 17 September 2015
Localisation of RNAs and proteins in nucleolar precursor bodies of early mouse embryos 
Elena Lavrentyeva, Kseniya Shishova, German Kagarlitsky and Olga Zatsepina

Nucleolar precursor bodies (NPBs) are mandatory entities of mammalian zygotes, but their relation to nucleolus-like bodies (NLBs) of fully-grown oocytes remains vague. Here, we show that NPBs, unlike NLBs, are impoverished for RNA and rRNA despite of both bodies contain rRNA-binding proteins. We suggest a scheme for NPB assembly and transformation to functional nucleoli that supposes utilization of oocyte-derived nucleolar proteins but mainly of nascent rRNAs.

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Published online 09 September 2015
Effects of testosterone and prolactin on steroidogenesis in post-ovulatory cumuli oophori and on in vitro oocyte fertilisation in the rat 
Agnieszka Starowicz, Jerzy Galas, Ma?gorzata Duda, Zbigniew Tabarowski and Maria Szo?tys

Could a supplement hold the key to understanding the mechanism of progesterone production by the postovulatory cumulus-oocyte complexes (COCs)? In the research various supplements influencing progesterone production by rat COCs containing unfertilised or fertilised oocytes have been tested in vitro, suggesting that the hormone mixture of prolactin and testosterone increased progesterone synthesis and oocyte fertilisability. This might prompt a wider research on the use of this mixture to increase the IVF success rates also in other species.

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Published online 08 September 2015
Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender 
Asmatullah Kaka, Wahid Haron, Rosnina Yusoff, Nurhusien Yimer, A. M. Khumran, Kazhal Sarsaifi, Atique Ahmed Behan, Ubedullah Kaka, Akeel Ahmed Memon and Mahdi Ebrahimi

Earlier studies have shown that in vitro supplementation of DHA produced inconsistent results on the quality of frozen–thawed bull semen. This work aimed to overcome the variations reported previously. Results showed that 3 ng mL–1 DHA supplemented in BioXcell extender improved all sperm parameters compared with the control and other groups. The successful incorporation of DHA into the cell membrane before freezing protected the spermatozoa from disintegration caused by ice crystals.

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Published online 07 September 2015
Follicle vascularity coordinates corpus luteum blood flow and progesterone production 
S. G. S. de Tarso, G. D. A. Gastal, S. T. Bashir, M. O. Gastal, G. A. Apgar and E. L. Gastal

Over the last decade important advances have been made in the area of ovarian function using color-Doppler ultrasonography. Herein we investigated the influence of dimensions and vascularity of the preovulatory follicle (POF) wall as indicators to predict the size, blood flow, and capacity of progesterone production of the subsequent CL in cows. Strong correlations and linear associations between POF and/or CL dimensions and blood flow with systemic progesterone concentrations from the resultant CL were observed. Selection of POFs with greater blood flow is likely to result in CLs with improved potential for plasma progesterone production.

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Published online 04 September 2015
Ultrastructure of spermatogenesis and spermatozoa in agoutis during sexual development 
M. A. M. Arroyo, F. F. S. Silva, P. R. S. Santos, A. R. Silva, M. F. Oliveira and A. C. Assis Neto

The ultrastructure of the spermatogenesis in agoutis help to identify the processes that affect infertility and the use of biotechnologies related to reproduction. The Leydig and Sertoli cells undergo morpho-physiological changes during sexual development, especially in the prepubertal phase. The spermatozoon of agouti had a tapered head without prominence of the acrosome and no evidence of the perforatorium. There is evidence of degeneration of centrioles in completely formed spermatozoa. In order to deepen the knowledge on spermatogenesis of agouti kept in captivity, this work highlights the importance of reproduction in wild species for their preservation and rational creation.

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Published online 27 August 2015
Swim-up of tammar wallaby (Macropus eugenii) spermatozoa in Biggers, Whitter and Whittingham (BWW) medium: maximisation of sperm motility, minimisation of impairment of sperm metabolism and induction of sperm hyperactivation 
Minjie Lin, Xiyi Zhang, Ray N. Murdoch and R. John Aitken

For the first time in a marsupial species, this study visually revealed that spermatozoa of the tammar wallaby can be quickly induced into a hyperactivated pattern, a necessary condition that enables spermatozoa to fertilise eggs in nature. The study also identified that BWW medium is able to maintain sperm motility, metabolism and viability better than other commonly used media. As there has been no success with IVF in any marsupials so far, this information could enable researchers to develop a proper IVF protocol for endangered marsupial species.

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Published online 27 August 2015
Kinetic effect of oestrogen on secretion of prostaglandins E2 and F2? in bovine oviduct epithelial cells 
Zhiheng Dong, Nan Zhang, Wei Mao, Bo Liu, Na Huang, Peifeng Li, Changyou Li and Jinshan Cao

Prostaglandins are important factors in mediating fertilisation and establishing pregnancy. In this study we investigate the effect of oestrogen on prostaglandin E2 (PGE2) and prostaglandin F2α (PGF) secretion and the role of synthase on PGE2 and PGF2a production in bovine oviduct epithelial cells. The findings suggest that exogenous oestradiol, PGE2 and PGF may modify bovine oviduct reproductive endocrine dysfunction.

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Published online 25 August 2015
Differential gene expression in the endometrium reveals cytoskeletal and immunological genes in lactating dairy cows genetically divergent for fertility traits 
Bruce Moran, Stephen T. Butler, Stephen G. Moore, David E. MacHugh and Christopher J. Creevey

We examined the global gene expression profile of endometrial tissue on Day 7 of the oestrous cycle in Holstein cows with good or poor genetic merit for fertility traits. Three physiologically relevant networks of genes were identified: (1) actin and cytoskeletal components; (2) immune function; and (3) ion transportation. This study has identified three important areas for future research.

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    | Supplementary Material (120 KB)
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Published online 25 August 2015
Combinatorial effects of epidermal growth factor, fibroblast growth factor 2 and insulin-like growth factor 1 on trophoblast cell proliferation and embryogenesis in cattle 
Ming Xie, Sarah R. McCoski, Sally E. Johnson, Michelle L. Rhoads and Alan D. Ealy

In vitro embryo culture is gaining popularity in various species as a means for generating pregnancies. In this study, supplementing culture medium with uterine-derived factors improved bovine placental cell proliferation and embryo development. These findings support the contention that providing selective uterine-derived factors during early embryo culture improves embryo competency for maintaining viable pregnancies and producing healthy offspring.

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    | Supplementary Material (118 KB)
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Published online 25 August 2015
Dizygotic monochorionic canine fetuses with blood chimaerism and suspected freemartinism 
Carolynne J. Joonè, Kurt G. M. De Cramer and Johan O. Nöthling

The presence of two canine fetuses within one placental site has been rarely documented in the literature. In this case report, we describe the finding of monochorionic canine fetuses of discordant gender, with co-mingling of the fetal blood supplies. Abnormal sexual development was discovered in both fetuses, which, in the female, resembles freemartinism, a phenomenon not previously reported in this species.

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Published online 25 August 2015
Transgenerational effects of a hypercaloric diet 
A. O. Joaquim, C. P. Coelho, P. Dias Motta, E. F. Bondan, E. Teodorov, M. F. M. Martins, T. B. Kirsten, R. C. V. Casarin, L. V. Bonamin and M. M. Bernardi

The maternal diet can alter the bodyweight and immune programming of subsequent generations. A maternal hypercaloric diet (HD) during puberty and early adult age induces overweight in the F1 generation and increases peripheral inflammation despite the feeding of a normocaloric diet to the F1 generation. It also induces adaptative patterns of the hypothalamic glial response towards a state of neuroprotection. These data reveal a transgenerational effect of the HD that may occur, in part, through permanent changes in immune and nervous system programming.

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    | Supplementary Material (155 KB)
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Published online 24 August 2015
Effect of seminal plasma proteins on the motile sperm subpopulations in ram ejaculates 
Carolina Luna, Marc Yeste, María M. Rivera del Alamo, Juan Domingo, Adriana Casao, Joan E. Rodriguez-Gil, Rosaura Pérez-Pé, José A. Cebrián-Pérez and Teresa Muiño-Blanco

Seminal plasma proteins (SPP) support survival of ram spermatozoa, exerting a dual effect, both capacitating and decapacitating. The addition of SPP before in vitro capacitation with epidermal growth factor induced a decrease in progressive spermatozoa and an increase in hyperactivated spermatozoa. Ram spermatozoa are able to undergo capacitation with no hyperactivation and SPP are able to induce hyperactivation in spermatozoa but maintain them in a decapacitated state.

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Published online 21 August 2015
Effect of C-type natriuretic peptide on maturation and developmental competence of immature mouse oocytes in vitro 
Qiang Wei, Chuan Zhou, Min Yuan, Yuyang Miao, Xiaoe Zhao and Baohua Ma

Being able to induce meiotic arrest, C-type natriuretic peptide (CNP) provides a new alternative to improve the synchronisation of nuclear and cytoplasmic maturation of oocytes matured in vitro. In the present study, temporary meiotic arrest induced by CNP significantly improved maturation and the subsequent development of mouse immature oocytes in vitro. These results indicate the potential value of CNP in improving the efficiency of oocyte IVM.

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Published online 21 August 2015
Nuclear transfer alters placental gene expression and associated histone modifications of the placental-specific imprinted gene pleckstrin homology-like domain, family A, member 2 (PHLDA2) in cattle 
Daniel R. Arnold, Roberta C. Gaspar, Carlos V. da Rocha, Juliano R. Sangalli, Tiago H. C. de Bem, Carolina A. P. Corrêa, João C. T. Penteado, Flavio V. Meirelles and Flavia L. Lopes

Comparing placental development of bovine embryos produced naturally, by IVF and by cloning would allow for improvements in animal production using assisted reproductive techniques. The present study demonstrates that gene expression of the imprinted gene pleckstrin homology-like domain, family A, member 2 (PHLDA2) is higher in early stage cloned placentas. This increase in PHLDA2 mRNA is associated with changes in the epigenetic event of histone methylation.

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Published online 21 August 2015
Effect of intra-uterine growth restriction on long-term fertility in boars 
Yan Lin, Xu Cheng, Peter Sutovsky, De Wu, Lian-Qiang Che, Zheng-Feng Fang, Sheng-Yu Xu, Bo Ren and Hong-Jun Dong

The reproductive performance of offspring in adulthood is determined during fetal and neonatal development. The aim of the present study was to explore the effect of intra-uterine growth restriction on the reproductive performance of offspring. Growth restriction affected sperm production via reproductive organ development and epigenetic regulation. These results provide evidence for the effects of prenatal development on programming reproductive performance in male offspring.

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Published online 21 August 2015
Selection of reference genes for quantitative real-time polymerase chain reaction in porcine embryos 
Won-Jae Lee, Si-Jung Jang, Seung-Chan Lee, Ji-Sung Park, Ryoung-Hoon Jeon, Raghavendra Baregundi Subbarao, Dinesh Bharti, Jeong-Kyu Shin, Bong-Wook Park and Gyu-Jin Rho

The gene expression pattern varies between porcine embryos produced by different methods and to analyse these changes normalisation against reference genes is routinely employed. However, these reference genes should be carefully chosen for evaluation as they also tend to have variable expression. In our study using three different analysis programs, we determined that SDHA and H2A are the most-stable reference genes that can be used for accurate normalisation to obtain reliable results.

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Published online 14 August 2015
Changes in aquaporin 5 in the non-ciliated cells of mouse oviduct according to sexual maturation and oestrous cycle 
Won Heum Nah, Yeong Seok Oh, Jung Hye Hwang and M. C. Gye

Aquaporins (AQPs) play an important role in fluid homeostasis in epithelia. To understand oviductal fluid homeostasis, AQP5 expression was examined in immature and cycling female mice. In oviduct epithelia, AQP5 in non-ciliated cells was increased by gonadotropin in immature mice and peaked during oestrus in cycling females, suggesting its participation in fluid homeostasis in oviduct epithelia.

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Published online 11 August 2015
Puberty onset is delayed following uteroplacental insufficiency and occurs earlier with improved lactation and growth for pups born small 
Tania Romano, Deanne H. Hryciw, Kerryn T. Westcott and Mary E. Wlodek

Growth restriction increases the risk of developing disease in adulthood, with the postnatal environment playing a critical role. This study identified the effects of prenatal and postnatal growth restriction on puberty onset and metabolism in growth restricted rats. The results show that growth restricted males and females had a delayed puberty onset, with altered sex-hormone concentrations and reduced leptin. Improvement of postnatal nutrition leads to a restoration of plasma leptin concentrations and rectification of the timing of puberty onset.

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Published online 11 August 2015
Time within reproductive season, but not age or inbreeding coefficient, affects seminal and sperm quality in the whooping crane (Grus americana) 
M. E. Brown, S. J. Converse, J. N. Chandler, A. L. Crosier, W. Lynch, D. E. Wildt, C. L. Keefer and N. Songsasen

The whooping crane underwent a severe genetic bottleneck in the early 1940s. The present study demonstrates that time of breeding season, but not inbreeding coefficient, affects the seminal characteristics of cranes maintained ex situ. The findings will serve as an important foundation for efforts in incorporating AI with frozen–thawed semen into genetic and demographic management of this endangered species.

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Published online 03 August 2015
Follicular size and stage and gonadotropin concentration affect alginate-encapsulated in vitro growth and survival of pre- and early antral dog follicles 
Jennifer Nagashima, David E. Wildt, Alexander J. Travis and Nucharin Songsasen

This study aims to understand the complex dynamics of dog folliculogenesis through in vitro studies. This study demonstrates that both preantral and early antral stage follicle grow significantly in vitro with FSH, but not LH, being a key regulator. The findings serve as important foundation for the establishment of in vitro follicle culture system for rescuing maternal genomes in domestic and wild canids.

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Published online 29 July 2015
Asynchronous embryo transfer as a tool to understand embryo–uterine interaction in cattle: is a large conceptus a good thing? 
Federico Randi, Beatriz Fernandez-Fuertes, Michael McDonald, Niamh Forde, Alan K. Kelly, Habner Bastos Amorin, Eduardo Muniz de Lima, Fabio Morotti, Marcelo Marcondes Seneda and Patrick Lonergan

Embryo–uterine synchrony is critical for the establishment of pregnancy. The aim was to examine the effect of embryo–uterine synchrony on conceptus development and pregnancy in cattle. Results demonstrate the strong regulatory effect of the uterus on conceptus development and the importance of synchrony for optimal pregnancy rates.

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Published online 21 July 2015
Assessment of yellowtail kingfish (Seriola lalandi) as a surrogate host for the production of southern bluefin tuna (Thunnus maccoyii) seed via spermatogonial germ cell transplantation 
Ido Bar, Andre Smith, Erin Bubner, Goro Yoshizaki, Yutaka Takeuchi, Ryosuke Yazawa, Ben Nan Chen, Scott Cummins and Abigail Elizur

Germ cell transplantation is an innovative technology for the production of interspecies surrogates, with the aim here of facilitating easy handling, shorter generation time and a more economical management of bluefin tuna broodstock. The present study demonstrated that transplanted southern bluefin tuna testicular germ cells migrate and colonise the gonads of yellowtail kingfish host, but do not further differentiate and proliferate.

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    | Supplementary Material (37 KB)
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Published online 21 July 2015
Effects of reduced glutathione on acrosin activity in frozen–thawed boar spermatozoa 
Efrén Estrada, Joan E. Rodríguez-Gil, Maria M. Rivera del Álamo, Alejandro Peña and Marc Yeste

Reduced glutathione (GSH) improves the fertilising ability of frozen–thawed boar spermatozoa. The present study investigated the relationship between acrosin activity and reproductive performance in frozen–thawed boar spermatozoa and the effects of GSH. The results show that GSH counteracts the decrease in acrosin activity in good but not poor freezability ejaculates and suggest that the increase in reproductive performance mediated by GSH could be related to acrosin activity.

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Published online 21 July 2015
Evidence that fibroblast growth factor 10 plays a role in follicle selection in cattle 
A. C. S. Castilho, C. A. Price, F. Dalanezi, R. L. Ereno, M. F. Machado, C. M. Barros, B. G. Gasperin, P. B. D. Gonçalves and J. Buratini

The mechanisms underlying follicle selection in monovular species are largely unknown. We assessed the involvement of FGF7 subfamily members in follicle selection in cattle and found that expression of ligands and receptors is upregulated in subordinate vs dominant follicles during follicle deviation, and that FGF10 inhibits oestradiol secretion and expression of FSHR and IGF1R in granulosa cells. These data point to an important role for FGF10 in follicle selection.

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Published online 21 July 2015
Mitogen-activated protein kinase-activated protein kinase 2 is a critical regulator of pig oocyte meiotic maturation 
Xiang-Hong Ou, Sen Li, Bao-Zeng Xu, Lei-Ning Chen, Man-Xi Jiang, Shao-Qin Chen and Nan-Qiao Chen

Oocyte quality is critical for female reproduction. Using a pig oocyte model, this study revealed the important role of mitogen-activated protein kinase-activated kinase (MK2) in regulating meiotic resumption and spindle organization, as well as euploid oocyte production. This will contribute to understanding the causes of female infertility and improving the success of assisted reproduction.

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Published online 16 July 2015
Factors supporting long-term culture of bovine male germ cells 
Mahesh Sahare, Sung-Min Kim, Ayagi Otomo, Kana Komatsu, Naojiro Minami, Masayasu Yamada and Hiroshi Imai

Bovine stem cells in the testis are destined to be spermatozoa in nature, but can be reprogrammed into pluripotent stem cells in culture. We describe the development of a new culture system for bovine spermatogonial stem cells that maintains their germ-cell and pluripotent-cell characteristics. This culture system provides a feasible strategy for establishing pluripotent germ-cell lines useful for genetic modification in domestic species and preservation of endangered species.

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Published online 16 July 2015
Retinoic acid treatment alters germ cell heterogeneity and testicular echotexture in prepubescent ram lambs 
Jennifer L. Giffin, Ann C. Hahnel and Pawel M. Bartlewski

Scrotal ultrasonography can be employed to study testicular development but echotextural characteristics of specific germ cell types remain unknown. Retinoic acid (RA) given to prepubescent ram lambs reduced the variation in germ cell populations, which corresponded with a significant change in numerical pixel values (brightness) of the testicular parenchyma. RA manipulation of spermatogenic function, particularly in sexually immature animals, may provide a useful model for further investigations into the non-invasive ultrasonographic monitoring of spermatogenesis.

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Published online 16 July 2015
Effects of birthweight on reproductive system development and onset of puberty in gilts 
F. R. C. L. Almeida, B. Laurenssen, L. X. Pereira, K. J. Teerds and N. M. Soede

Studies on subsequent reproductive performance of low birth weight gilts are limited. The effects of birth weight on the development of the reproductive tract and puberty onset were investigated herein, and both parameters seem equivalent in different birth weight gilts. As birth weight may be an important parameter to include in dam line breeding programs, a better understanding of the impact of birth weight on fertility seems critical.

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Published online 15 July 2015
Distribution pattern of cytoplasmic organelles, spindle integrity, oxidative stress, octamer-binding transcription factor 4 (Oct4) expression and developmental potential of oocytes following multiple superovulation 
Guruprasad Kalthur, Sujith Raj Salian, Ramya Nair, Jemey Mathew, Satish Kumar Adiga, Sneha Guruprasad Kalthur, Dimphy Zeegers and M. Prakash Hande

Due to the low success rate and poor ovarian response, female partners of infertile couple are often subjected to repeated superovulation irrespective of the type of infertility. Repeated exposures to supraphysiological levels of hormones raise concerns about the safety of this procedure on the general health of the individual, gamete quality and impact on embryonic development. In the present study, safety of repeated superovulation cycles was investigated by assessing the quality of oocytes and embryos obtained from mice subjected to repeated superovulation with a gap of 7 days between each cycle. Repeated superovulation was found to adversely affect the quality of oocytes as characterized by altered distribution of cytoplasmic organelles, spindle abnormalities, elevated oxidative stress and altered Oct-4 expression which was associated with decrease in the developmental potential of embryos.

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Published online 08 July 2015
Developmental potential of 2n/3n mixoploid mouse embryos produced by fusion of individual second polar bodies and blastomeres of 2-cell embryos 
Toshiaki Hino and Hiroyuki Tateno

Incorporation of a second polar body (PB2) with a blastomeres is considered as a causal mechanism of human 2n/3n mixoploidy. Using a mouse model, we produced 2n/3n mixoploid embryos to demonstrate their developmental competence. These embryos will be useful for understanding the production mechanism of human 2n/3n mixoploidy.

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    | Supplementary Material (71 KB)
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Published online 06 July 2015
Fertilisation of cryopreserved sperm and unfertilised quail ovum by intracytoplasmic sperm injection 
Kyung Soo Kang, Tae Sub Park, Deivendran Rengaraj, Hyung Chul Lee, Hong Jo Lee, Hee Jung Choi, Shusei Mizushima, Tamao Ono and Jae Yong Han

Intracytoplasmic sperm injection (ICSI) is a promising tool for the conservation of animal genetic resources, but remains a challenge in birds. In this study we demonstrated that cryopreserved spermatozoa combined with ICSI followed by surrogate eggshell culture can result in fertilisation and embryo development in the quail. This study could provide technical improvements to maintain avian genetic diversity using ICSI.

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Published online 06 July 2015
Effect of stallion age on the expression of LH and FSH receptors and aromatase P450 in equine male reproductive tissues 
Carlos Vladimir Herrera-Luna, Dragos Scarlet, Ingrid Walter and Christine Aurich

Although stallions may reach an age of >20 years, their fertility often starts to decline earlier. This study demonstrates differences in the responsiveness of testicular tissue to LH and FSH between prepubertal and mature stallions. In postpubertal stallions, no further changes occurred with increasing age. Aromatase, a key enzyme for oestrogen synthesis, seems to be of importance for the maintenance of fertility.

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Published online 06 July 2015
High fructose consumption in pregnancy alters the perinatal environment without increasing metabolic disease in the offspring 
Christopher Lineker, Paul M. Kerr, Patricia Nguyen, Ian Bloor, Stuart Astbury, Nikhil Patel, Helen Budge, Denise G. Hemmings, Frances Plane, Michael E. Symonds and Rhonda C. Bell

Maternal dietary intake is important for a healthy pregnancy but it is not clear whether high intake of sugar, particularly fructose, has long-term adverse effects on the offspring. Mothers fed fructose were fatter, had mild diabetes, and poor vascular control in pregnancy; offspring did not receive fructose after birth and were heavier at 3 months but not different from control offspring at 6 months of age. Raised fructose during pregnancy resulted in poor maternal health but no major detrimental effects in offspring up to 6 months of age.

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Published online 01 July 2015
Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants 
D. Acha, M. Hidalgo, I. Ortiz, M. J. Gálvez, J. J. Carrasco, V. Gómez-Arrones and J. Dorado

A suitable method for cryopreserving semen is needed for the conservation of endangered donkey breeds. This study compared the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. It concluded that Gent B improved in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% ethylene glycol or 2.5% dimethylformamide increased sperm protection against cryodamage.

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Published online 30 June 2015
Ovarian suppression in a marsupial following single treatment with a gonadotrophin-releasing hormone agonist in microspheres 
Ryan Robert Witt, Ian Ross Forbes, John McBain and John Cameron Rodger

A practical female synchronisation strategy for marsupials is needed if artificial insemination using frozen stored spermatozoa or other assisted breeding methods are to be part of conservation breeding programs. This study investigated the potential for Lucrin Depot, a therapeutic drug used to suppress reproductive activity in humans, to regulate the marsupial cycle. The results show that Lucrin Depot suppresses reproductive activity and has the potential to synchronise the recovery of breeding activity in the fat-tailed dunnart.

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Published online 29 June 2015
Morphometric analysis and developmental comparison of embryos from carriers with balanced chromosomal rearrangements in preimplantation genetic diagnosis cycles 
Baoheng Gui, Zhongyuan Yao, Yanru Huang, Libin Mei, Yanping Li, Donge Liu, Nenghui Liu, Yan Xia, Desheng Liang and Lingqian Wu

Embryos from carriers with balanced chromosomal rearrangements show distinct developmental characteristics. This study quantified embryo morphological parameters and demonstrates that morphometric characteristics are correlated with developmental outcomes as well as with chromosomal composition in embryos from carriers with balanced chromosomal rearrangements. Combining morphometric and qualitative assessment may increase the precision of embryo evaluation and contribute to improving the efficiency of preimplantation genetic diagnosis.

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    | Supplementary Material (61 KB)
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Published online 26 June 2015
Testicular injection of busulfan for recipient preparation in transplantation of spermatogonial stem cells in mice 
Yusheng Qin, Ling Liu, Yanan He, Wenzhi Ma, Huabin Zhu, Mingyuan Liang, Haisheng Hao, Tong Qin, Xueming Zhao and Dong Wang

Transplantation of spermatogonial stem cells (SSCs) has theoretical and practical significance in discovering the mechanisms underlying spermatogenesis and increasing male reproduction potential. We developed a method to prepare recipients for SSCs transplantation using testicular injections of busulfan. The recipients did not die with this method and normal donor offspring were obtained. This safe method could be adapted for use in other species in the future.

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Published online 25 June 2015
Reproductive hormones affect follicular cells and ooplasm of Stage I and II oocytes in zebrafish 
Maria Lígia Sousa, Ana Silva, Fernanda Malhão, Maria João Rocha, Eduardo Rocha and Ralph Urbatzka

Little is known about the regulation of follicle development at the first stages (I and II) in fish or the potential role of hormones. Reproductive hormones had different effects on the ooplasm (increased presence of endoplasmic reticulum) and the granulosa­–theca cell layer (reduced progression of cell cycle) of zebrafish follicles, predominantly at Stage II. These results deliver insights into the follicle development of fish species with an asynchronous mode of reproduction.

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    | Supplementary Material (193 KB)
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Published online 25 June 2015
Effects of brain-derived neurotrophic factor on oocyte maturation and embryonic development in a rat model of polycystic ovary syndrome 
Qiaoli Zhang, Dong Liu, Meiling Zhang, Na Li, Shulan Lu, Yanzhi Du and Zi-Jiang Chen

Polycystic ovary syndrome (PCOS) is one of the most common reproductive endocrine disorders. It causes abnormal ovulation and infertility in women. We established a rat PCOS model and demonstrated that appropriate concentrations of brain-derived neurotrophic factor (BDNF) promoted oocyte maturation and embryonic development in this model. Our studies indicate that BDNF may benefit women with PCOS by promoting oocyte developmental capacity and embryonic formation.

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    | Supplementary Material (397 KB)
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Published online 18 June 2015
Steroids affect gene expression, ciliary activity, glucose uptake, progesterone receptor expression and immunoreactive steroidogenic protein expression in equine oviduct explants in vitro 
Hilde Nelis, Bartosz Wojciechowicz, Anita Franczak, Bart Leemans, Katharina D'Herde, Karen Goossens, Pieter Cornillie, Luc Peelman, Ann Van Soom and Katrien Smits

Hormones play a key role during the oestrous cycle and pregnancy of the mare, but little information is available on their influence at the level of the oviduct. We used an in vitro model to establish the effect of hormones on the morphology, functionality and gene expression of the equine oviduct. The findings can be implemented in further studies on embryo–maternal interactions.

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Published online 15 June 2015
Lactation-induced changes in metabolic status and follicular-fluid metabolomic profile in postpartum dairy cows 
Niamh Forde, Aoife O'Gorman, Helena Whelan, Pat Duffy, Lydia O'Hara, Alan K. Kelly, Vitezslav Havlicek, Urban Besenfelder, Lorraine Brennan and Pat Lonergan

Poor fertility in lactating dairy cows is a worldwide phenomenon, which has serious financial implications, particularly in season systems of production. This study characterised lactation-induced differences in the composition of the follicular fluid, which potentially impact negatively on the developing oocyte. These data will contribute to our understanding of the factors affecting fertility and may lead to strategies to address them.

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Published online 15 June 2015
Extended ex vivo culture of fresh and cryopreserved whole sheep ovaries 
Sara Maffei, Giovanna Galeati, Georgia Pennarossa, Tiziana A. L. Brevini and Fulvio Gandolfi

It is difficult to study the mammalian ovary in vitro because it is a complex organ that requires a three-dimensional structure to sustain follicle development and oocyte maturation. This study describes an original perfusion system for the culture of whole ovine ovaries for up to 4 days preserving follicle development and endocrine function. This system provides a relevant model for physiological and toxicological studies.

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Published online 05 June 2015
Androgen and oestrogen modulation by D-aspartate in rat epididymis 
S. Falvo, M. M. Di Fiore, L. Burrone, G. Chieffi Baccari, S. Longobardi and A. Santillo

In the testes D-aspartate induces testosterone synthesis, but its action in the epididymis is unknown. The effects of in vivo D-aspartate administration in rat epididymis were investigated. The findings show that D-aspartate modulates androgen and oestrogen levels and expression of their receptors by acting on 5α-reductase and P450-aromatase gene expression. Our findings support the role of D-aspartate in the male reproductive system.

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Published online 03 June 2015
Interaction of the transforming growth factor-? and Notch signaling pathways in the regulation of granulosa cell proliferation 
Xiao-Feng Sun, Xing-Hong Sun, Shun-Feng Cheng, Jun-Jie Wang, Yan-Ni Feng, Yong Zhao, Shen Yin, Zhu-Mei Hou, Wei Shen and Xi-Feng Zhang

Do signalling pathways in organisms work independently or dependently? To understand the precise mechanism of cell proliferation in follicular development, we conducted this study and found that synergistic effects of the transforming growth factor (TGF)-β and Notch signalling pathways play an important role in regulating granulosa cell proliferation. The interaction between the TGF-β and Notch signalling pathways found in granulosa cells may also exist in many other types of cells.

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Published online 29 May 2015
Plasma prolactin concentrations during lactation, pouch young development and the return to behavioural oestrus in captive koalas (Phascolarctos cinereus) 
K. Ballantyne, S. T. Anderson, A. Mucci, V. Nicolson and S. D. Johnston

Increased prolactin (PRL) levels during lactation in the koala are regarded as a key factor contributing to female anoestrus. In this study, PRL concentrations in captive lactating koalas remained low until 161–170 days after parturition, at which time concentrations increased significantly. Given that behavioural oestrus was suppressed throughout the majority of lactation, elevated PRL levels do not appear to be the cause of oestrous cycle suppression.

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Published online 27 May 2015
Activation of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor gene expression following DNA demethylation in placental choriocarcinoma and transformed cell lines 
Pan-Hong Wu, Xue-Mei Chen, Xue-Qing Liu, Jun-Lin He, Qian Feng, Xi Lan, Xue Zhang, Yan-Qing Geng, Ying-Xiong Wang and Yu-Bin Ding

DNA methylation and gene expression of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors are differed from choriocarcinoma and transformed trophoblastic cell lines. And we showed DNA methylation is associated with TRAIL receptor expression and might be involved in trophoblast apoptosis.

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    | Supplementary Material (340 KB)
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Published online 26 May 2015
Head morphology of ram spermatozoa is associated with their ability to migrate in vitro and correlates with fertility 
Carmen Martínez-Rodríguez, Mercedes Alvarez, Elena López-Urueña, Susana Gomes-Alves, Luis Anel-López, Jorge E. Tizado, Luis Anel and Paulino de Paz

The head morphology of spermatozoa that migrated in a penetration test was evaluated. The relationships among head morphometric characteristics, the ability to penetrate the cervical mucus surrogate and fertility were evaluated. In conclusion, the proportion of spermatozoa with a short and wide head in the fresh sperm sample may be used as a negative predictor of male fertility.

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    | Supplementary Material (304 KB)
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Published online 21 May 2015
Trolox enhances follicular survival after ovarian tissue autograft in squirrel monkey (Saimiri collinsi) 
Sarah R. Scalercio, Christiani A. Amorim, Danielle C. Brito, Sandro Percário, Irma C. Oskam, Sheyla F. S. Domingues and Regiane R. Santos

Ovarian tissue transplantation remains risky due to ischaemic–reperfusion injury leading to follicular degeneration or massive activation of follicles from the reserve pool. As oxidative stress might be involved in such a loss, we report herein the use of the antioxidant Trolox, which was able to enhance follicular survival and to avoid extreme follicular activation in grafted ovarian tissue from squirrel monkeys.

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Published online 20 May 2015
Neonatal testosterone exposure induces early development of follicular cysts followed by sympathetic ovarian hyperinnervation 
Gabriel Anesetti and Rebeca Chávez-Genaro

Elevated neonatal androgens act as disturbers of follicular ovarian growth, inducing the formation of cysts in the adult ovary. In this work we analysed the temporal correlation between cyst development and changes in sympathetic ovarian innervation. An increase in density of innervation after cyst development was observed; this could be associated with an attempt to restore normal ovarian activity or as a sign of loss of ovarian functionality.

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Published online 20 May 2015
Genomic expression profiles in cumulus cells derived from germinal vesicle and MII mouse oocytes 
Li Shao, Ri-Cheng Chian, Yixin Xu, Zhengjie Yan, Yihui Zhang, Chao Gao, Li Gao, Jiayin Liu and Yugui Cui

Cumulus cells (CCs) are distinct from other granulosa cells surrounding the egg and the expression of some genes in CCs during maturation may predict oocyte quality. In this study, some genes exhibited differential expression in CCs and could be potential biomarkers for egg development following fertilisation. The expression of these genes could be used for oocyte selection in the fields of animal husbandry and clinical infertility treatment.

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Published online 20 May 2015
Effect of ovarian hormones on the healthy equine uterus: a global gene expression analysis 
Christina D. Marth, Neil D. Young, Lisa Y. Glenton, Drew M. Noden, Glenn F. Browning and Natali Krekeler

For successful reproduction, the uterus needs to be finely regulated by hormones to adapt to its various functions. The aim of this study was to generate gene expression profiles for two cycle stages in the equine uterus, which identified major changes in the expression of genes associated with its morphology, immune system and metabolic processes. Understanding these physiological variations helps distinguish and identify pathologies.

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    | Supplementary Material (2.8 MB)
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Published online 19 May 2015
Influence of bovine serum albumin and fetal bovine serum supplementation during in vitro maturation on lipid and mitochondrial behaviour in oocytes and lipid accumulation in bovine embryos 
Maite del Collado, Naiara Z. Saraiva, Flavia L. Lopes, Roberta C. Gaspar, Luciana C. Padilha, Roberta R. Costa, Guilherme F. Rossi, Roberta Vantini and Joaquim M. Garcia

Lipids and mitochondria are essential to oocyte and embryo energy metabolism; however, their behaviour during in vitro production of bovine embryos remains unknown. We demonstrated the effects of media supplements on lipid and mitochondrial behaviour during in vitro maturation and their impact on embryo lipid accumulation. Variations in mitochondrial and lipid dynamics caused by in vitro maturation may have negative effects on oocyte development and embryo lipid accumulation.

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Published online 18 May 2015
Involvement of G9A-like protein (GLP) in the development of mouse preimplantation embryos in vitro 
Xian-Ju Huang, Xueshan Ma, Xuguang Wang, Xiaolong Zhou, Juan Li, Shao-Chen Sun and Honglin Liu

G9A-like protein (GLP) plays an important role in mouse early embryonic development and Glp-deficient embryos exhibit severe growth retardation and defects that lead to lethality at approximately Embryonic Day 9.5. In this study we investigated the effect of Glp deficiency on in vitro early embryonic development. Knockdown of Glp impaired the pluripotency of mouse embryos. The results of the present study may be applicable to human prenatal diagnosis.

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    | Supplementary Material (474 KB)
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Published online 15 May 2015
Subtraction suppressive hybridisation analysis of differentially expressed genes associated with puberty in the goat hypothalamus 
G. L. Cao, T. Feng, M. X. Chu, R. Di, Y. L. Zhang, D. W. Huang, Q. Y. Liu, W. P. Hu and X. Y. Wang

Discovery of genes associated with the onset of puberty is important for precocious goat breeds. The aim of the present study was to find the genes associated with the onset of puberty in the goat. The results indicate that energy production and conversion may be initiating factors for puberty in the goat. The genes associated with these processes can be validated and may be used in goat breeding.

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    | Supplementary Material (116 KB)
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Published online 14 May 2015
Adoptive transfer of transforming growth factor-?1-induced CD4+CD25+ regulatory T cells prevents immune response-mediated spontaneous abortion 
Tian Qiu, Yincheng Teng, Yudong Wang and Liang Xu

T cells induced with anti-CD3, anti-CD28, IL-2, and TGF-β1 could prevent spontaneous abortion in mice. Adoptive transfer of CD4+CD25+ or TGFB1-induced Treg cells significantly reduced the fetal resorption rate, increased serum IL10 and TGF-β1 levels, and decreased IFNG levels. Autologous transfer of in vitro TGF-β1-induced Treg cells may represent a novel therapeutic strategy for women with unexplained recurrent spontaneous abortions.

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    | Supplementary Material (70 KB)
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Published online 13 May 2015
In vitro culture of stem-like cells derived from somatic cell nuclear transfer bovine embryos of the Korean beef cattle species, HanWoo 
Daehwan Kim, Sangkyu Park, Yeon-Gil Jung and Sangho Roh

Bovine somatic cell nuclear transfer embryo-derived stem-like cells were established under defined culture conditions containing three inhibitors of differentiation. Long-term maintenance of stemness and differentiation capacity of the cells was demonstrated. The results have increased our understanding of the state of bovine pluripotent stem cells (PSCs) and provides new insights into the establishment of PSCs in domestic animals.

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    | Supplementary Material (890 KB)
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Published online 13 May 2015
Comparison of different fertilisation media for an in vitro maturation–fertilisation–culture system using flow-cytometrically sorted X chromosome-bearing spermatozoa for bovine embryo production 
Luis B. Ferré, Yanina Bogliotti, James L. Chitwood, Cristóbal Fresno, Hugo H. Ortega, Michael E. Kjelland and Pablo J. Ross

The aim of this study was to evaluate three fertilisation media (M199, synthetic oviductal fluid (SOF) and Tyrode’s albumin–lactate–pyruvate (TALP)) on IVF performance using female sex-sorted spermatozoa. Embryos derived from SOF and TALP media had higher numbers of ICM, TE and total cells than those fertilised in M199. The fertilisation medium used affected cleavage rate and subsequent embryo development, quality and hatching ability.

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Published online 08 May 2015
Relocalisation and activation of integrins induced rapidly by oestrogen via G-protein-coupled receptor 30 in mouse blastocysts 
Ting Qu, Shi-mao Zhang, Lin-lin Yu, Sheng Zhang, Dong-Zhi Yuan, Qian Xu, Jin-Hu Zhang, Ya-ping He and Li-min Yue

It is necessary for estrogen to activate embryos at blastocysts stage during implantation. In this research, we explored the mechanism of estrogen action onblastocysts and found estrogen could rapidly cause the relocalisation of integrins in blastocyst cells, and enhanced their affinity with the ligands, so that they attached and implanted into uterine endometrium. Our results suggest a possibility of improving outcome of in vitro fertilisation (IVF).

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Published online 08 May 2015
Association of a region of bovine chromosome 1 (BTA1) with age at puberty in Angus bulls 
María E. Fernández, Alberto Prando, Andrés Rogberg-Muñoz, Pilar Peral-García, Andrés Baldo, Guillermo Giovambattista and Juan P. Lirón

Age at puberty is an important trait for reproductive performance of cattle, so the identification of genes regulating this character could assist in the selection for early pubertal animals. Through an association analysis in Angus bulls, two candidate genes for puberty were detected. These results could contribute to the understanding of puberty regulation and advanced research in the context of reproduction.

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    | Supplementary Material (669 KB)
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Published online 06 May 2015
Characterisation of the deleted in azoospermia like (Dazl)–green fluorescent protein mouse model generated by a two-step embryonic stem cell-based strategy to identify pluripotent and germ cells 
Priscila Ramos-Ibeas, Eva Pericuesta, Raúl Fernández-González, Alfonso Gutiérrez-Adán and Miguel Ángel Ramírez

The deleted in azoospermia like (Dazl) gene was initially described as a master regulator of germline specification; however, recent studies indicate that it may have pluripotency-related functions. We generated Dazl–green fluorescent protein-transgenic mice in which the transgene is expressed in both pluripotent and germ cells. These transgenic mice represent a valuable tool for the prospective identification of stem cells from several tissues.

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    | Supplementary Material (38 KB)
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Published online 05 May 2015
Transcriptome of early embryonic invasion at implantation sites in a murine model 
J. M. Moreno-Moya, N. A. Franchi, S. Martínez-Escribano, J. A. Martínez-Conejero, S. Bocca, S. Oehninger and J. A. Horcajadas

Successful implantation relies on the interaction between a competent embryo and a receptive endometrium. The aim of the present study was to investigate genes specifically expressed in early invasive embryonic tissue versus decidual tissue using microarray analysis. Genes differentially expressed in the maternal compartment included chemokines, lipoproteins, growth factors and transcription factors, whereas the embryonic invasive tissue expressed genes commonly observed in invasive tumour-like processes. The results provide information about genes involved in early embryonic invasion and the control exerted by the surrounding decidua.

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    | Supplementary Material (105 KB)
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Published online 05 May 2015
Potential role of maternal lineage in the thoroughbred breeding strategy 
Xiang Lin, Shi Zhou, Li Wen, Allan Davie, Xinkui Yao, Wujun Liu and Yong Zhang

Results of two association studies indicate that maternal heritability is a stronger contributor to race performance than paternal heritability in a sample of thoroughbred horses in Australia. In addition, 8–11 years appeared to be the optimal age range of dams for child bearing in relation to their offsprings’ achievements in racing. These findings warrant further research into the significance and contribution of maternal heritability in breeding practice for thoroughbred racehorses.

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Published online 05 May 2015
Dynamics of Notch signalling in the mouse oviduct and uterus during the oestrous cycle 
D. Murta, M. Batista, A. Trindade, E. Silva, L. Mateus, A. Duarte and L. Lopes-da-Costa

The oviductal and uterine cellular remodelling occurring along the estrous cycle (EC) requires a finely tuned intercellular communication. We found that the Notch pathway is enrolled in this cell signalling. Notch genes are differentially transcribed and expressed in the oviduct and uterus along the EC, and this is correlated with progesterone concentrations. Overall, the study implies the Notch pathway as a regulator of the cellular remodelling occurring along the EC.

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Published online 05 May 2015
Multiple roles of hypoxia in ovarian function: roles of hypoxia-inducible factor-related and -unrelated signals during the luteal phase 
Ryo Nishimura and Kiyoshi Okuda

After the discovery of hypoxia-inducible factor-1 (HIF1), the importance of low oxygen conditions (hypoxia) in physiological responses has become recognized. This review focuses on the roles of hypoxia in ovarian physiology, especially during luteal phase. Because the reports concerning the roles of hypoxia in ovarian function are increasing, the understanding of this area will help further understanding of ovarian function.

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Published online 05 May 2015
Relationship between seminal plasma tuberoinfundibular peptide of 39 residues and sperm functional attributes in buffalo (Bubalus bubalis) 
Sellappan Selvaraju, Lakshminarayana Somashekar, Binsila B. Krishnan, Sivashanmugam Parthipan, Guvvala Pushparani, Arunachalam Arangasamy, Duraisamy Rajendran and Janivara Parameswaraiah Ravindra

Assessment of semen quality is important to improve fertility in the buffalo. The aim of this study was to assess the seminal plasma protein profile in relation to semen function. We found that tuberoinfundibular peptide of 39 residues (TIP39) protein in seminal plasma was important for maintaining sperm function and fertilising ability. TIP39 could be a potential molecule to assess semen quality in buffalo bulls.

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    | Supplementary Material (29 KB)
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Published online 05 May 2015
Novel equine conceptus–endometrial interactions on Day 16 of pregnancy based on RNA sequencing 
C. Klein

Maintenance of pregnancy is dependent on the exchange of signals between the conceptus and the endometrium. This is the first report of concurrent transcriptome analysis of conceptus and endometrium in the mare. A plethora of interactions between genes expressed by the conceptus and endometrium 16 days after ovulation in the horse were revealed, providing rationale for further investigation.

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Published online 01 May 2015
The use of a synthetic progesterone, levonorgestrel (LNG), to control the oestrous cycle in the koala 
K. Ballantyne, S. T. Anderson, M. Pyne, V. Nicolson, A. Mucci, A. Lisle and S. D. Johnston

Obtaining control of the koala reproductive cycle in order to manipulate the timing of oestrus is a fundamental component of artificial insemination in this species. Synthetic progesterone implants were used to temporarily suppress oestrous cycle activity in female koalas. Implant removal resulted in the synchronous return to oestrus in three of the four treated animals. These results provide hope for the effective use of a progesterone-based oestrus synchronisation tool in the koala.

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Published online 01 May 2015
Use of the gonadotrophin-releasing hormone antagonist azaline B to control the oestrous cycle in the koala (Phascolarctos cinereus) 
K. Ballantyne, S. T. Anderson, M. Pyne, V. Nicolson, A. Mucci, A. Lisle and S. D. Johnston

Administration of a GnRH antagonist for the purposes of oestrous synchronization in a marsupial species is reported for the first time. Injections of azaline B temporarily inhibited the LH response in koalas to exogenous mGnRH and successfully delayed the return to oestrus. Although azaline B clearly disrupts koala folliculogenesis, further investigation and refinement is required to implement this pharmaceutical into an effective oestrous synchronisation protocol.

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Published online 30 April 2015
Comparative genomic identification and expression profiling of a novel ?-defensin gene cluster in the equine reproductive tract 
Gillian P. Johnson, Andrew T. Lloyd, Cliona O'Farrelly, Kieran G. Meade and Sean Fair

β-defensins are small cationic proteins that have been shown to play a pivotal reproductive–immunobiological role across a range of species including mice, macaques and men. This study profiles a cluster of 13 novel β-defensin genes along Chromosome 22 of the Equus caballus genome and characterises their expression in the reproductive tract of the stallion and mare. β-defensin preferential regional expression in the caput epididymis in the stallion and the oviduct in the mare suggests a possible role in immunoprotection of the equine reproductive tract.

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Published online 22 April 2015
Changes in lysosomal enzymes and mannose-6-phosphate receptors related to sexual maturation in bull epididymis 
Andrea C. Aguilera, Lorena Carvelli, Verónica Boschin, Fabián Mohamed, Leila Zyla and Miguel A. Sosa

Mammalian epididymides secrete proteins into the lumen, which contribute to sperm maturation. The aim of the present study was to investigate the activity and distribution of lysosomal enzymes in bull epididymides at two different reproductive ages. It was found that enzyme activity and distribution vary during sexual maturity in bull epididymides and are regulated by mannose-6-phophate receptors. These findings could provide some molecular basis for male infertility.

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Published online 22 April 2015
MicroSort® sperm sorting causes no increase in major malformation rate 
Donald P. Marazzo, David Karabinus, Lawrence A. Johnson and Joseph D. Schulman

MicroSort sperm sorting is a groundbreaking technology that increases the odds of having a child of the indicated sex. The present study compared the rates of major malformations in pregnancies conceived with or without the use of MicroSort technology and demonstrated no increase in the rate of major malformations. This study represents a model for evaluating a crucial aspect of safety related to reproductive interventions in humans.

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Published online 22 April 2015
Spindle configuration and developmental competence of in vitro-matured bovine oocytes exposed to NaCl or sucrose prior to Cryotop vitrification 
Núria Arcarons, Roser Morató, Jozé F. W. Spricigo, Marcia A. M. M. Ferraz and Teresa Mogas

Bovine oocytes are particularly difficult to cryopreserve successfully. In the present study we examined whether exposure of in vitro-matured bovine oocytes to high concentrations of NaCl or sucrose before vitrification and warming would improve their cryotolerance. Pretreatment with 375 mOsmol NaCl or sucrose had no adverse effects on the spindle status of vitrified–warmed cow oocytes. However, sucrose pretreatment offered no benefits for embryo development.

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Published online 21 April 2015
Spatio-specific regulation of endocrine-responsive gene transcription by periovulatory endocrine profiles in the bovine reproductive tract 
Estela R. Araújo, Mariana Sponchiado, Guilherme Pugliesi, Veerle Van Hoeck, Fernando S. Mesquita, Claudia M. B. Membrive and Mario Binelli

Early embryo mortality hinders fertility in domestic species and humans, and is associated with poor maternal receptivity. Receptivity depends on sex-hormone priming of the maternal tissue to result in balanced secretions of nutrients and growth factors for the embryo. Here, we showed that abundance of molecules important for embryo receptivity differs according to the region of the reproductive tract. This provides the molecular basis for the characterisation of microenvironments within the reproductive tract that are associated with embryo receptivity and fertility.

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Published online 21 April 2015
Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2? and PGE2 production in the bovine endometrium 
Mariko Kuse, Ryosuke Sakumoto and Kiyoshi Okuda

Endometrial prostaglandins (PGs) are the most important modulators of the oestrous cycle and are regulated by ovarian hormones. This study aimed to clarify the regulatory mechanisms of progesterone on PG production. Our findings demonstrate that progesterone has at least two actuating pathways and precisely controls PG production by regulating enzymes of the arachidonate cascade via genomic pathways and by controlling signal transduction via non-genomic pathways.

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Published online 21 April 2015
Sperm superoxide dismutase is associated with bull fertility 
Kamilah E. Grant, Rodrigo V. de Oliveira, Bettye Sue Hennington, Aruna Govindaraju, Andy Perkins, John Stokes, Dennis Rowe, Einko Topper, Abdullah Kaya, Arlindo Moura and Erdogan Memili

Fertility is the most important factor regulating animal reproduction. The aim of the present study was to ascertain to what extent sperm proteins β-tubulin 2C, heat shock protein 10 and superoxide dismutase 1 (SOD1) were associated with bull fertility using molecular and bioinformatics approaches. We demonstrated that SOD1 was associated with in vivo bull fertility and that it interacts with key regulators of cell viability. The findings are significant because they reveal the paternal effect protein involved in bull fertility.

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Published online 20 April 2015
Subclinical mastitis disrupts oocyte cytoplasmic maturation in association with reduced developmental competence and impaired gene expression in preimplantation bovine embryos 
Z. Roth, S. Asaf, O. Furman, Y. Lavon, D. Kalo, D. Wolfenson and G. Leitner

Subclinical mastitis is associated with reduced fertility in dairy cows; however, the underlying mechanism is not entirely clear. Herein we provide evidence, for the first time, that subclinical mastitis induces changes in oocyte maturation in association with reduced developmental competence. Effects were observed at both the cellular and molecular levels following the induction of subclinical mastitis using toxins of Staphylococcus aureus (Gram-positive) or Escherichia coli (Gram-negative) origin.

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Published online 17 April 2015
Expression of leukaemia inhibitory factor at the conceptus–maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare 
M. de Ruijter-Villani, C. Deelen and T. A. E. Stout

Leukaemia inhibitory factor (LIF) is essential for murine blastocyst survival and implantation. We demonstrate that LIF expression increases at the conceptus–maternal interface in the mare when the trophectoderm and endometrium first come into direct contact, i.e. as the intervening blastocyst capsule disintegrates. Since LIF receptor expression in the trophoblast increases simultaneously, we propose that LIF contributes to trophoblast development and non-invasive implantation in horses.

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Published online 17 April 2015
Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques 
J. Dorado, M. J. Gálvez, S. Demyda-Peyrás, I. Ortiz, J. M. Morrell, F. Crespo, J. Gósalvez and M. Hidalgo

The use of chilled semen has a special importance in canine breeding; however, the quality of chilled dog semen decreases during cold storage. This study compared the efficacy of simple sperm washing (SW), single-layer centrifugation (SLC) and modified swim-up (SU) techniques in the preparation of dog spermatozoa for cooling. It was concluded that SU through Androcoll-C can be integrated into canine semen preservation protocols.

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Published online 15 April 2015
C-Type natriuretic peptide maintains domestic cat oocytes in meiotic arrest 
Yougang Zhong, Jiabao Lin, Xiaoping Liu, Jian Hou, Yong Zhang and Xingxu Zhao

C-Type natriuretic peptide (CNP) plays an essential role in maintaining meiotic arrest of mouse and porcine oocytes. Herein we studied the effect of CNP on spontaneous maturation of oocytes from the domestic cat and found that CNP is required to maintain oocyte meiotic arrest and prevent oocyte degeneration in this species. Therefore, CNP seems ubiquitously involved in the meiotic arrest of oocytes in mammals.

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Published online 15 April 2015
miR-424/322 is downregulated in the semen of patients with severe DNA damage and may regulate sperm DNA damage 
Kai Zhao, Yaoping Chen, Ruifeng Yang, Yang Bai, Cuiling Li, Honggang Li and Chengliang Xiong

This study explored the relationship between testis miRNAs and sperm DNA damage. We found that miR-424/322 was significantly downregulated in semen of patients with severe DNA damage and its downregulated expression in mouse testis increased sperm DNA damage. This is, therefore, a novel mechanism of sperm DNA damage and potential therapeutic target.

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Published online 15 April 2015
Potential effects of interferon regulatory factor 4 in a murine model of polyinosinic-polycytidylic acid-induced embryo resorption 
Jing Wang, Tailang Yin, Yanqi Wen, Fuju Tian, Xiaojun He, Danni Zhou, Yi Lin and Jing Yang

How pathogens lead to pregnancy failure remains unclear. To explore signal transduction in the effects of pathogens, a mouse model of pathogen-induced embryo loss was established. Pathogen-produced double-stranded RNA may activate an immune response via interferon regulatory factor 4, subsequently leading to embryo loss. The findings of the present study may be useful in preventing or treating infection-related pregnancy failure.

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Published online 13 April 2015
Hypoxia limits mouse follicle growth in vitro 
J. M. Connolly, M. T. Kane, L. R. Quinlan, P. Dockery and A. C. Hynes

There are many problems with trying to grow or culture follicles and eggs outside the human body for reproductive therapies. Even with current best practice, success rates of 10% or less are commonplace. We show that paying particular attention to the level of oxygen in the culture environment can greatly improve follicle growth.

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Published online 31 March 2015
Epigenetic pattern changes in prenatal female Sprague-Dawley rats following exposure to androgen 
Yanjie Xia, Shanmei Shen, Xinlin Zhang, Zhantao Deng, Zou Xiang, Hongwei Wang, Long Yi, Qian Gao and Yong Wang

Polycystic ovary syndrome (PCOS), a common cause of female infertility, may be predetermined in the womb. Exposing mothers to excess amounts of an androgenic hormone may render their female offspring susceptible to developing PCOS due to altered regulation of androgen production-related genes in the offspring. This research has furthered our understanding of the mechanisms of PCOS and sheds light on disease prevention.

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Published online 19 March 2015
Different locations of RANTES and its receptors on mouse epididymal spermatozoa 
Jin-Hua Wei, Xiao Feng, Zhi-Jian Sun, Pang Cheng, Bin-Fang Ma, Jie Zhao, Yu-Hang Dong, Yuan-Qiang Zhang and Zhen Li

The present study was undertaken to investigate the association between the chemokine RANTES and epididymal spermatozoa. RANTES and its receptors were not in the same sperm location, suggesting that if RANTES binds to mouse epididymal spermatozoa, it would be independent of the receptors. Our study provides a new basis for further investigations of the association of RANTES with spermatozoa.

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Published online 19 March 2015
The employment of IVF techniques for establishment of sodium, copper and selenium impact upon human sperm quality 
Urszula Marzec-Wróblewska, Piotr Kami?ski, Pawe? ?akota, Marek Szyma?ski, Karolina Wasilow, Grzegorz Ludwikowski, Magdalena Kuligowska-Prusi?ska, Gra?yna Odrow??-Sypniewska, Tomasz Stuczy?ski and Alina Wo?niak

The levels of trace elements in spermatozoa and whole semen may significantly affect male fertility. The aim of this study was to analyse the levels of Na, Cu and Se in human semen and GPx activity in seminal plasma: Na, Cu, Se and GPx are related to sperm characteristics and male fertility and their survey could improve male infertility diagnosis.

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Published online 18 March 2015
Effects of local testicular heat treatment on Leydig cell hyperplasia and testosterone biosynthesis in rat testes 
Zhonghai Li, Jianhai Tian, Genggang Cui, Meng Wang and Dapeng Yu

Rather little attention has been paid to the molecular mechanisms in the effects of heat treatment on Leydig cell function. In this study, testicular heat treatment led to Leydig cell hyperplasia and a reduction in testosterone levels. The findings of this study provide new insights into improvements in the treatment of male infertility.

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Published online 17 March 2015
Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts 
Y. H. Choi, M. C. T. Penedo, P. Daftari, I. C. Velez and K. Hinrichs

Detection of sex, disease status and other genetic traits of horse embryos by biopsy before they are transferred would be a valuable breeding management tool. We found that accuracy differed between methods used for DNA amplification, but not between in vivo- and in vitro-produced embryos. The final high accuracy achieved (>95%) using DNA amplification and multiplex PCR supports clinical use of this technique.

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Published online 13 March 2015
Developmental competence of 8–16-cell stage bison embryos produced by interspecies somatic cell nuclear transfer 
L. Antonio González-Grajales, Laura A. Favetta, W. Allan King and Gabriela F. Mastromonaco

Interspecies nuclear transfer can be a valuable tool for the preservation of threatened wildlife species; however, numerous challenges remain. Development of embryos reconstructed from wood bison donor cells transplanted into enucleated domestic cattle oocytes was affected as early as the 8–16-cell stage with altered cellular function and viability. These findings provide evidence for low blastocyst development rates and highlight the influence of interspecies communication in reconstructed embryos.

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Published online 13 March 2015
Sperm rescue in wild African elephants 
Joseph Saragusty, María Teresa Prieto, Alexandre Courtiol, Romain Potier, Frank Göritz, Thomas B. Hildebrandt and Robert Hermes

What do you do when you have large volume of frozen semen from an endangered species with quality too poor to be used in assisted reproductive programs? Facing such a conundrum we tested a number of density-gradient centrifugation media, finding Percoll as the medium of choice for wild African elephant spermatozoa. Rather than discarding semen of inferior quality, density-gradient centrifugation can help select for better-quality and useable spermatozoa.

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Published online 13 March 2015
Relationship between sperm apoptosis and bull fertility: in vivo and in vitro studies 
Lauren Erickson, Tom Kroetsch and Muhammad Anzar

Early assessment of fertility is important for bull selection in a breeding program. The relationships of sperm membrane and DNA fitness with bull fertility were determined. Spermatozoa with membrane and DNA breaks in bull semen adversely affected field fertility following artificial insemination. Cow ova fertilised with semen possessing DNA-nicked spermatozoa demonstrated poor embryonic development in the laboratory.

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Published online 10 March 2015
Human sperm aneuploidy after exposure to polycyclic aromatic hydrocarbons 
Michał Radwan, Joanna Jurewicz, Wojciech Sobala, Sławomir Brzeźnicki, Paweł Radwan, Lucjusz Jakubowski, Wanda Hawuła, Anna Ulańska and Wojciech Hanke

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in daily life and have endocrine-disrupting properties that may affect reproductive and developmental processes. The purpose of this study was to investigate whether 1-hydroxypyrene – a biomarker of PAH exposure – was associated with sperm aneuploidy. The results suggest that exposure to PAHs may be associated with sperm aneuploidy. As this is the first such epidemiological study these findings require further replication.

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Published online 10 March 2015
Steroids in the equine oviduct: synthesis, local concentrations and receptor expression 
Hilde Nelis, Julie Vanden Bussche, Bartosz Wojciechowicz, Anita Franczak, Lynn Vanhaecke, Bart Leemans, Pieter Cornillie, Luc Peelman, Ann Van Soom and Katrien Smits

Early pregnancy influences hormones in the blood of the mare, but little is known about local hormone concentrations in the reproductive tract, which is in direct contact with the developing equine embryo. We determined hormone levels in the oviduct of the horse and our main finding was that the concentration of the pregnancy hormone progesterone is significantly elevated in the oviduct after ovulation and on the side of ovulation. This study provides the basis for further research into embryo–maternal interactions in the equine oviduct.

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Published online 05 March 2015
The impact of storage temperature and sperm number on the fertility of liquid-stored bull semen 
Craig Murphy, Shauna A. Holden, Edel M. Murphy, Andrew R. Cromie, Patrick Lonergan and Sean Fair

The use of liquid bull semen in artificial insemination provides an ideal opportunity to maximise the number of inseminations per ejaculate through a lower number of spermatozoa per insemination dose. This study demonstrated that a temperature range of 5 to 22°C is not detrimental to liquid sperm function and that lowering the number of spermatozoa per liquid insemination dose from 5 to 3 million does not result in a significant decline in fertility. This reduction in sperm number allows for the better use of genetically superior sires leading to enhanced genetic gain.

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Published online 27 February 2015
Effects of long-term dietary supplementation with conjugated linoleic acid on bovine oocyte lipid profile 
Andrés F. González-Serrano, Christina R. Ferreira, Valentina Pirro, Andrea Lucas-Hahn, Julia Heinzmann, Klaus-Gerd Hadeler, Ulrich Baulain, Patrick Aldag, Ulrich Meyer, Marion Piechotta, Gerhard Jahreis, Sven Dänicke, R. Graham Cooks and Heiner Niemann

Nutritional and environmental conditions around conception and during early embryonic development have significant effects on health and well-being in adult life. Here, we used a bovine heifer model to investigate the effects of rumen-protected fat supplementation on oocyte quality and embryo development with the aid of advanced mass spectrometry and molecular methods. Results show that rumen-protected fatty-acid supplementation affects oocyte lipid content and may pave the way towards a better understanding of reproductive disorders associated with nutritional impairments.

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    | Supplementary Material (753 KB)
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Published online 25 February 2015
Spermatozoa of Sminthopsis murina (Mammalia: Metatheria) exhibit an unusually high degree of chromatin stability in the absence of disulphide bonding in protamine 1 
S. D. Johnston, C. López-Fernández, F. Arroyo, S. Fardell, R. Roy and J. Gosálvez

Despite a lack of cysteine in the protamine fraction of dunnart spermatozoa, the chromatin remains remarkably stable. Using a range of molecular techniques and under conditions known to fragment sperm DNA, we further confirm this phenomenon. Our results suggest that cysteine is present in the histone fraction of the chromatin and it may be functioning to conform tertiary and/or quaternary chromatin structure.

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Published online 24 February 2015
Association between endometrial thickness in oocyte donation cycles and pregnancy success rates 
Hans Arce, Esther Velilla and Marisa López-Teijón

Endometrial receptivity is a primary concern in fertility treatments. No significant differences in pregnancy rate, the number of gestational sacs and miscarriage rates were found with different endometrial thickness measurements in the present study, the largest study so far evaluating the role of endometrial thickness in oocyte donation cycles. Hence, embryo transfer can take place once endometrial thickness >5 mm has been observed in a single ultrasonographic evaluation, with no need for subsequent monitoring.

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Published online 24 February 2015
Oestrous cycle-dependent expression of Fas and Bcl2 family gene products in normal canine endometrium 
S. Henriques, E. Silva, S. Cruz, M. F. Silva, G. Ferreira-Dias, L. Lopes-da-Costa and L. Mateus

Knowledge of the molecular mechanisms regulating apoptosis in normal canine cyclic endometrium might lead to a better understanding of tissue changes associated with implantation and uterine pathological conditions such as cystic endometrial hyperplasia. The results of our work indicate that pro- and anti-apoptotic proteins regulate the balance between cell proliferation and death in the canine endometrium during the oestrous cycle.

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Published online 23 February 2015
Gene expression in the mammary gland of the tammar wallaby during the lactation cycle reveals conserved mechanisms regulating mammalian lactation 
C. J. Vander Jagt, J. C. Whitley, B. G. Cocks and M. E. Goddard

The tammar wallaby has evolved a unique lactation strategy compared to eutherian mammals making it a valuable comparative model for lactation studies. In this study, the tammar wallaby mammary gland was investigated for changes in gene expression during key stages of the lactation cycle and differentially regulated genes were identified. These genes may play a crucial regulatory role in marsupial milk production, and are likely to play a related role in other mammals.

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Published online 19 February 2015
Expression and immunolocalisation of follicle-stimulating hormone receptors in gonads of newborn and adult female horses 
Dragos Scarlet, Ingrid Walter, Juraj Hlavaty and Christine Aurich

FSH and its receptor (FSHR) are essential for the development and function of the ovary. This study demonstrates for the first time a constant expression and immunolocalisation of FSHR in ovaries from adult mares and newborn fillies, together with the existence of tertiary follicles up to 4 mm in female neonates. Our results suggest an involvement of FSH in luteal function and oocyte maturation in horses.

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Published online 18 February 2015
Use of an organotypic mammalian in vitro follicle growth assay to facilitate female reproductive toxicity screening 
Yuanming Xu, Francesca E. Duncan, Min Xu and Teresa K. Woodruff

Reproductive toxicity screening relies on animal testing. We applied our in vitro follicle growth assay to screen for endocrine and gamete outcomes following exposure to compounds including an oil dispersant used heavily during the Deepwater Horizon oil spill. Here we show that in vitro follicle health can be monitored in vitro, which may help inform future human health exposure policy.

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Published online 18 February 2015
An integrated approach to bovine oocyte quality: from phenotype to genes 
Leslie Angulo, Catherine Guyader-Joly, Sylvain Auclair, Christelle Hennequet-Antier, Pascal Papillier, Mekki Boussaha, Sébastien Fritz, Karine Hugot, François Moreews, Claire Ponsart, Patrice Humblot and Rozenn Dalbies-Tran

In cattle, a suboptimal pool of oocyte factors is suspected to contribute to the poor reproductive performance. Cows with high and low oocyte developmental potential in vitro were discriminated, and differential transcripts evidenced using microarrays; reverse transcription–quantitative polymerase chain reactionconfirmed altered expression for SLC25A16, PPP1R14C, ROBO1, AMDHD1 and MEAF6. These genes will be examined for polymorphisms that could be used in genomic selection.

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Published online 18 February 2015
Mutations in the leptin receptor gene associated with delayed onset of puberty are also associated with decreased ovulation and lambing rates in prolific Davisdale sheep 
Jennifer L. Juengel, Michelle C. French, Anne R. O'Connell, Sara J. Edwards, Avijit Haldar, Rudiger Brauning, Phil A. Farquhar, Ken G. Dodds, Susan M. Galloway, Peter D. Johnstone and George H. Davis

Reproductive efficiency is controlled by genetics and environmental factors such as nutrition. Using a sheep model, we have discovered that mutations in the leptin receptor, a key pathway in the nutritional control of reproduction, are associated with reduced fertility. This has potential applications for the selection of sheep with improved reproductive efficiency as well as providing a new model to better understand how leptin regulates fertility in mammals.

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Published online 16 February 2015
Food deprivation and social inequality may lead to oxidative damage: a study on the preventive role of melatonin in the male rat reproductive system 
Shiva Nasiraei-Moghadam, Kazem Parivar, Abolhasan Ahmadiani, Mansoureh Movahhedin and Mohammad-Reza Vaez Mahdavi

Spermatogenic cells are susceptible to oxidative stress. Food deprivation and the sense of inequality in food-deprived rats that see other rats have access to normal food (hereafter referred to as ‘instability’) are known stressors that can increase oxidative stress. In this study, food deprivation and instability increased markers of oxidative stress, whereas melatonin treatment and isolation of rats (which balanced instability) decreased these markers.

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Published online 10 February 2015
Morphological and molecular characterisation of Twitcher mouse spermatogenesis: an update 
Erica Puggioni, Laura Governini, Martina Gori, Giuseppe Belmonte, Paola Piomboni, Elvira Costantino-Ceccarini and Alice Luddi

Altered sphingolipid metabolism has been suggested to cause sperm morphological abnormalities in Twitcher, a mouse model of Krabbe disease. However, our data demonstrate that a derangement of the hypothalamic–pituitary–gonadal axis, caused by severe brain degeneration, may contribute to Twitcher infertility. These findings may also contribute in further understanding of all cases of infertility associated with neurological impairment.

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Published online 09 February 2015
Effects of ovarian hyperstimulation on mitochondria in oocytes and early embryos 
Jing Shu, Li-Li Xing, Guo-Lian Ding, Xin-Mei Liu, Qing-Feng Yan and He-Feng Huang

Millions of babies come from artificial reproductive technology, which harvests eggs commonly by superovulation. Is this safe? We studied the superovulation effects on mice egg mitochondria, the important organelle reflecting egg potential and with genetic function. The results showed a decline in the average mitochondrial quality by superovulation. However, it is not superovulation that causes the damage, it is superovulation that recruits more eggs with poor mitochondrial quality instead.

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Published online 15 January 2015
Breed-specific factors influence embryonic lipid composition: comparison between Jersey and Holstein 
Luis Baldoceda, Isabelle Gilbert, Dominic Gagné, Christian Vigneault, Patrick Blondin, Christina Ramires Ferreira and Claude Robert

Dairy cattle breeds differ considerably in their milk production performance but breed effects are seldom considered for reproductive performance. It is known that Jersey embryos display poor post-freezing survival rates compare to Holstein. We demonstrate that Jersey embryos contain more lipid droplets associated with a lower mitochondrial activity and also that the lipid composition between the two breeds is different. Even under same management conditions, breed can influence the embryos lipid content and metabolism.

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Published online 14 January 2015
Frequency of exogenous hormone therapy impacts spermiation in male Fowler 
Caitlin E. McDonough, Michelle W. Martin, Carrie K. Vance, Judith A. Cole and Andrew J. Kouba

Amphibians are experiencing a global extinction crisis and captive assurance colonies in zoos and aquariums are unsustainable over the long-term due to poor reproduction. In this study, we aimed to improve our understanding of the use of hormone therapy in male toads for sperm collection. Our results have clear implications that frequent hormone-induced spermiation could have a negative effect on sperm production and we recommend that hormone injections be spaced a minimum of 2 weeks apart to optimise the health of the animals and provide optimum sperm quality for assisted reproductive technologies.

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Published online 14 January 2015
The effects of glucuronic acid and N-acetyl-D-glucosamine on in vitro fertilisation of porcine oocytes 
K. Schmidt, A. Clark, A. Mello, C. Durfey, A. Buck, K. Boyd and B. D. Whitaker

Reducing the incidence of polyspermic penetration may aid in improving embryonic development and further the understanding of fertilisation in pigs. This study reveals that by supplementing oocytes with perivitelline space components, the space thickness increases and polyspermic penetration is reduced. The findings highlight the need for adequate environments to improve the maturation of oocytes prior to fertilisation.

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Published online 13 January 2015
Maternal treatment with fluoxetine promotes testicular alteration in male rat pups 
Aline C. Ramos, Alice H. dos Santos, Kennia M. Silveira, Ana Carolina I. Kiss, Suzana F. P. Mesquita and Daniela C. C. Gerardin

Previous results obtained in our laboratory showed that maternal exposure to fluoxetine (FLX) alters testicular morphometric parameters in adult male offspring. The present study investigated the effects of maternal FLX exposure on maternal behavior and on testes function offspring in the weanling and pubertal periods in rats. FLX treatment did not impair mother–pup interactions; however, it could impair testicular function in offspring.

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Published online 13 January 2015
Genetic influence on the reduction in bovine embryo lipid content by l-carnitine 
Luis Baldoceda, Dominic Gagné, Christina Ramires Ferreira and Claude Robert

Holstein and Jersey embryos display difference in term of lipid profile and molecular characteristics, suggesting a variance in lipid metabolism. Using L-carnitine, we noticed a reduction in embryonic lipid content for both breeds, but the response to treatment is weaker in the Jersey. L-carnitine supplementation reduces lipid content of blastomere and offers the potential to improve cryopreservation success.

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Published online 07 January 2015
Characterisation of several ankyrin repeat protein variant 2, a phosphoprotein phosphatase 1-interacting protein, in testis and spermatozoa 
Joana Vieira Silva, Luís Korrodi-Gregório, Georg Luers, Maria João Cardoso, António Patrício, Nuno Maia, Edgar F. da Cruz e Silva and Margarida Fardilha

Phosphoprotein phosphatase 1 (PPP1) is essential for male fertility and the key to understanding PPP1 function lies in the identification of its interacting partners. This study was undertaken to characterise several ankyrin repeat protein variant 2 (SARP2), a PPP1-interacting protein, highly enriched in testis and spermatozoa. Given the expression pattern of SARP2 and its association with PPP1 and sperm-associated antigen 9, we suggest that it plays a role in spermiogenesis and sperm function, namely in sperm motility and the acrosome reaction.

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Published online 07 January 2015
Expression and function of transcription factor AP-2γ in early embryonic development of porcine parthenotes 
Sung-Hyun Lee, Jung-Woo Kwon, Inchul Choi and Nam-Hyung Kim

This is the first study to report on the expression and functions of transcription factor activator protein (AP) 2γ in pig embryos. AP2γ was detected at all stages during preimplantation and depletion negatively affected early embryo development. These findings suggest that AP2γ may play a role in critical cellular events, including cell adhesion, proliferation and cell fate, during preimplantation development.

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Published online 07 January 2015
Prenatally administered dexamethasone impairs folliculogenesis in spiny mouse offspring 
Monika Hułas-Stasiak, Piotr Dobrowolski and Ewa Tomaszewska

Prenatal exposure to dexamethasone causes developmental alterations in the ovaries of offspring. This study demonstrates that dexamethasone administered during the fetal period enhances follicular atresia by activating programmed cell death, which, in turn, depletes the stock of ovarian follicles and can lead to reduction of fertility in adult females. The findings highlight the need for careful use of dexamethasone in pregnant women.

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Published online 07 January 2015
Gelatinases and their tissue inhibitors are associated with oxidative stress: a potential set of markers connected with male infertility 
Ewa M. Kratz, Anna Kałuża, Mirosława Ferens-Sieczkowska, Beata Olejnik, Renata Fiutek, Mariusz Zimmer and Agnieszka Piwowar

The search for new diagnostic markers of male infertility takes into account factors other than sperm characteristics. The expression of matrix metalloproteinase (MMP)-9 and MMP-2 may be regulated by oxidative stress in various pathophysiological processes. Thus, the presence of seminal MMPs, their tissue inhibitors (tissue-specific inhibitor of metalloproteinase (TIMP)-1, TIMP-2) and their correlation with levels of oxidative stress parameters (i.e. advanced oxidation protein products (AOPP)) is interesting in the context of male fertility. High AOPP and an increased MMP-9 : TIMP-1 ratio alters the oxidative–antioxidative balance of the ejaculate, thereby reducing male fertility. Thus, reducing male fertility, and therefore these parameters may serve as additional diagnostic markers of semen quality and male reproductive potential.

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Published online 07 January 2015
Cooled semen for fixed-time artificial insemination in beef cattle 
Juliana C. Borges-Silva, Márcio R. Silva, Daniel B. Marinho, Eriklis Nogueira, Deiler C. Sampaio, Luiz Orcírio F. Oliveira, Urbano G. P. Abreu, Gerson B. Mourão and Roberto Sartori

Oestradiol-based fixed-time artificial insemination (FTAI) programs have a relatively wide window of time for ovulation that may decrease pregnancy per AI (P AI–1) depending on semen processing. We have shown a 20% increase in P AI–1 in beef cattle when cooled versus frozen–thawed semen was used, because of better sperm quality and longevity.

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Published online 07 January 2015
Increased mRNA expression of selected pro-inflammatory factors in inflamed bovine endometrium in vivo as well as in endometrial epithelial cells exposed to Bacillus pumilus in vitro 
Martina A. Gärtner, Sarah Peter, Markus Jung, Marc Drillich, Ralf Einspanier and Christoph Gabler

Inflammatory processes in the bovine endometrium caused by bacterial infection often result in reduced fertility. We aimed to elucidate the first inflammatory response of the bovine endometrium. Increased mRNA expression of pro-inflammatory factors was connected to inflammation in vivo and was provoked in endometrial epithelial cells co-cultured with the potentially pathogenic Bacillus pumilus. These findings suggest that the endometrium can react promptly to bacterial infections.

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Published online 07 January 2015
Histochemical structure and immunolocalisation of the hyaluronan system in the dromedary oviduct 
Omnia Mohey-Elsaeed, Waleed F. A. Marei, Ali A. Fouladi-Nashta and Abdel-Aleem A. El-Saba

Hyaluronic acid is a major component of the extracellular matrix and is also found in body fluids. We show immunohistochemical localization of its synthases, main receptor and degrading enzymes in the camel oviduct. Regional differences in its synthesis and degradation suggest a differential role in the oviduct–gamete interaction in the ampulla and isthmus.

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Published online 05 January 2015
Actin crosslinking protein filamin A during early pregnancy in the rat uterus 
Romanthi J. Madawala, Connie E. Poon, Samson N. Dowland and Christopher R. Murphy

Filamin A is an actin-binding protein that was found to be associated with actin cytoskeletal remodelling of both luminal uterine epithelial cells (UECs) and endometrial stromal cells (ESCs) during early pregnancy. The distribution of filamin A in the endometrium was influenced by deciduogenic factors, while distribution of filamin A was found to be dependent on ovarian hormones. The expression of filamin A in the endometrium during early pregnancy contributes to the molecular and morphological changes that both UECS and ESCs undergo in order for them to be receptive to implantation by contributing to the remodelling of the actin cytoskeleton and hence the maintenance of plasma membrane stability of UECs and ESCs during early pregnancy.

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Published online 05 January 2015
Creation of DNA aptamers against recombinant bone morphogenetic protein 15 
Jun Sheng Lin, Alexia Kauff, Yong Diao, Huiyong Yang, Steve Lawrence and Jennifer L. Juengel

The oocyte-derived growth factor bone morphogenetic protein (BMP) 15 plays important roles in fertility, but its mechanism of action remains poorly understood. Sequences of single-stranded DNA specifically recognising BMP15 have been generated by a directed molecular evolutionary process. These special sequences (aptamers) can be used as a tool to detect BMP15 and decipher its biological actions.

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Published online 05 January 2015
Effects of dietary quercetin on female fertility in mice: implication of transglutaminase 2 
Kelly E. Beazley and Maria Nurminskaya

Increasing use of dietary supplementation with bioflavonoids such as quercetin raises concerns about their potential impacts on reproductive health. Here we show a negative impact of quercetin consumption on female fertility in the animal model and identify its target, transglutaminase 2, as a novel regulator of ovarian ageing. Thus the beneficial effects of quercetin may be overshadowed in females of reproductive age.

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Published online 17 December 2014
High oxygen tension increases global methylation in bovine 4-cell embryos and blastocysts but does not affect general retrotransposon expression 
Wenwen Li, Karen Goossens, Mario Van Poucke, Katrien Forier, Kevin Braeckmans, Ann Van Soom and Luc J. Peelman

Environmental stresses can have detrimental effects on the developing embryo that resonate throughout the rest of the life of that individual. In this study we tested whether the negative effects are induced by changes the chromatin structure and, if so, whether this loosens the brake on the activity of retrotransposons. Although global chromatin changes were observed, the results indicate that retrotransposons are also held in check by other factors.

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    | Supplementary Material (409 KB)
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Published online 17 December 2014
Impact of superovulation and mating on the wellbeing of juvenile and adult C57BL/6N mice 
Thomas Kolbe, Sarjoun Sheety, Ingrid Walter, Rupert Palme and Thomas Rülicke

Superovulation of mice is preferentially applied before sexual maturity of females is reached in order to maximize ova production. The effect of donor age in mice was evaluated with regard to distress induced by hormonal treatment and mating. Our results confirmed higher ovulation rates for immature females compared to adults and revealed no significant welfare issues for juvenile mice used for superovulation.

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    | Supplementary Material (85 KB)
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Published online 17 December 2014
Embryonic developmental plasticity in the long-snouted seahorse (Hippocampus reidi, Ginsburg 1933) in relation to parental preconception diet 
Francisco Otero-Ferrer, Marisol Izquierdo, Alireza Fazeli and William V. Holt

In seahorses, embryo development occurs within the male instead of the female, leading us to ask whether the male’s brood pouch has a similar function to the mammalian placenta. We specifically investigated whether the male’s diet before conception can influence the future survival and well being of newborn seahorses. Our experiment confirmed that to optimise the health and future survival of seahorses, male parents need to be supplied with dietary unsaturated fatty acids before they become pregnant.

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Published online 17 December 2014
Nutrition affects Sertoli cell function but not Sertoli cell numbers in sexually mature male sheep 
Yongjuan Guan, Guanxiang Liang, Penny A. R. Hawken, Sarah J. Meachem, Irek A. Malecki, Seungmin Ham, Tom Stewart, Le Luo Guan and Graeme B. Martin

A comprehensive evaluation of Sertoli cells has been used to investigate a reversible, non-pathological change in testis mass induced by nutrition in mature sheep. Under-nutrition did not affect Sertoli cell number but did affect Sertoli cell function, including reversal of cellular differentiation and disruption of tight-junctions. These responses were associated with major decreases in the output of spermatozoa.

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Published online 17 December 2014
Luteotropic and luteolytic factors regulate mRNA and protein expression of progesterone receptor isoforms A and B in the bovine endometrium 
Robert Rekawiecki, Magdalena Karolina Kowalik and Jan Kotwica

Expression of nuclear progesterone receptor isoforms A and B (PGRA and PGRB, respectively) can be regulated by various factors. PGRA and PGRB mRNA and protein expression was determined in bovine endometrium after stimulation with selected luteotropic and luteolytic factors. The data suggest that these factors affect PGRA and PGRB mRNA and protein levels and, in this way, regulate the effects of progesterone on endometrial cells.

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Published online 17 December 2014
New evidence of melatonin receptor contribution to ram sperm functionality 
Marta Gonzalez-Arto, Caorlina Luna, Rosaura Pérez-Pé, Teresa Muiño-Blanco, José A. Cebrián-Pérez and Adriana Casao

This study analysed the effects of melatonin through its receptors (MT1 and MT2) on ram sperm functionality. Using immunofluorescence assays and by evaluating the effects of MT1 and MT2 receptor agonists and antagonists, we determined that the modulating effect of melatonin on ram sperm capacitation seems to be receptor mediated, primarily via MT2 receptors. These results may explain the effects of exogenous melatonin on ram fertility.

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Published online 08 December 2014
Prospects for immunocontraception in feral horse population control: exploring novel targets for an equine fertility vaccine 
Aleona Swegen and R. John Aitken

Rapidly growing populations of feral horses can cause significant damage to landscapes and rural economies. Animal welfare concerns along with social and ethical considerations are increasingly precluding the use of lethal control methods to maintain acceptable population sizes, while non-lethal fertility control is not yet able to provide effective alternatives. This review explores the unique aspects of equine reproductive physiology that may provide opportunity for developing novel fertility vaccine strategies.

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Published online 08 December 2014
Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis 
Lalitha Devi, Himesh Makala, Lavanya Pothana, Khemlal Nirmalkar and Sandeep Goel

Testis cryopreservation is an efficient tool for preservation of the genetic potential of immature males that die before attaining sexual maturity. This is especially useful for buffalo calves that suffer a high mortality rate. Our results show that an optimum concentration of dimethylsulfoxide and fetal bovine serum could effectively preserve immature buffalo testicular tissues subjected to an uncontrolled slow-freezing protocol. Our findings encourage the cryo-banking of immature buffalo testicular tissues.

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Published online 04 December 2014
Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification 
Roser Morató, Míriam Castillo-Martín, Marc Yeste and Sergi Bonet

The ability to correctly select viable embryos is relevant to improving pregnancy rates and avoiding futile transfer attempts. Herein, we assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity prior to vitrification. Faster embryo development underlies higher blastocyst cryotolerance. The best results for survival after warming were obtained with the vitrification of porcine blastocysts presenting a high degree of expansion, particularly when vitrification was performed after 6 days of in vitro embryo culture .

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Published online 28 November 2014
Involvement of prolactin in the meloxicam-dependent inflammatory response of the gonadotropic axis to prolonged lipopolysaccharide treatment in anoestrous ewes 
Andrzej P. Herman, Agata Krawczyńska, Joanna Bochenek, Hanna Antushevich, Anna Herman and Dorota Tomaszewska-Zaremba

An immune challenge can affect the reproductive process in females. In this study, performed in ewes, we demonstrated that inhibition of cyclo-oxygenase 2 by meloxicam could minimise the negative effect of systemic inflammation induced by seven injections of bacterial endotoxin on gonadotropin-releasing hormone and LH secretion. However, the effectiveness of meloxicam treatment may be reduced during an inflammatory state characterized by high levels of prolactin.

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Published online 26 November 2014
Taking control of the female fertile lifespan: a key role for Bcl-2 family proteins 
Seng H. Liew, Kavitha Vaithiyanathan and Karla J. Hutt

The number of eggs laid down in the ovary at birth and subsequently maintained during reproductive life influences the length of the female fertile lifespan. Recent studies have revealed a critical role for cell death regulators belonging to the BCL-2 protein family in determining how many eggs are available to females to support fertility. This review summarises our current understanding of the proteins responsible for initiating and mediating cell death during the establishment and maintenance of the female germline.

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Published online 20 November 2014
Effect of spontaneous term labour on the expression of the NR4A receptors nuclear receptor related 1 protein (Nurr1), neuron-derived clone 77 (Nur77) and neuron-derived orphan receptor 1 (NOR1) in human fetal membranes and myometrium 
Martha Lappas

The upstream mechanisms involved in the processes of human labour are yet to be fully elucidated. This study shows that members of the nuclear receptor subfamily 4A (NR4A) are increased in fetal membranes and myometrium obtained from women after spontaneous term labour onset, and in the presence of various bacterial products. These findings suggest that NR4A family members may play a role in the processes of human labour including fetal membrane rupture and myometrial contractions.

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blank image Reproduction, Fertility and Development
Volume 28 Number 6 2016

 
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Suppressed expression of macrophage migration inhibitory factor in the oviducts of lean and obese cows 
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Asrafun Nahar and Hiroya Kadokawa
pp. 655-662

Oviducts synthesise macrophage migration inhibitory factor (MIF) to promote sperm capacitation and embryogenesis. We discovered that both obese and lean Japanese Black cows have suppressed expression of MIF mRNA and protein in the ampullae and isthmuses of their oviducts. Therefore, maternal nutritional condition may change expression of oviductal MIF, which plays important roles in fertilisation and early embryo development.

 
  
 

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Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality 
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Noelia Prieto-Martínez , Ingrid Vilagran , Roser Morató , Joan E. Rodríguez-Gil , Marc Yeste and Sergi Bonet
pp. 663-672

The aquaporin protein family allows the transport of water and other solutes through cell membranes and plays a role in reproductive physiology. The present work has studied, for the first time, the presence of AQP7 and AQP11 in boar spermatozoa. Although both were identified in this species, only AQP11 was found to be related to sperm quality.

 
  
 

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Endothelin as a local regulating factor in the bovine oviduct 
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Yuki Yamamoto , Misa Kohka , Yoshihiko Kobayashi , Izabela Woclawek-Potocka and Kiyoshi Okuda
pp. 673-681

Endothelin regulates oviductal motility, which is important for the transport of gametes and embryo. To define the endothelin system in the bovine oviduct, the production and target sites of endothelin were evaluated. All isoforms of endothelin are produced by epithelial cells and endothelin receptors localise in the smooth muscle, suggesting that the bovine oviduct possesses a local endothelin system.

 
  
 

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Leukotriene production profiles and actions in the bovine endometrium during the oestrous cycle 
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Anna J. Korzekwa , Robert Milewski , Martyna Łupicka and Dariusz J. Skarzynski
pp. 682-689

Leukotrienes are involved in reproductive functions in cows; however, their particular role in endometrium is unknown. The aim of the study was to elucidate the actions of leukotriene B4 and C4 in the bovine uterus. We demonstrated that these eicosanoids are produced, secreted and act in the endometrium depending upon the luteal stage. The results indicate multiple actions of leukotrienes in the bovine reproductive tract.

 
  
 

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Inhibition of angiopoietin-1 (ANGPT1) affects vascular integrity in ovarian hyperstimulation syndrome (OHSS) 
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Leopoldina Scotti , Dalhia Abramovich , Natalia Pascuali , Luis Haro Durand , Griselda Irusta , Ignacio de Zúñiga , Marta Tesone and Fernanda Parborell
pp. 690-699

OHSS is a disorder associated with ovarian stimulation and is characterised by ovarian enlargement and vascular permeability. The aim was to investigate the effect of follicular fluid from women at risk of OHSS, in the presence of an ANGPT1 inhibitor, on the chorioallantoic membrane assay and in endothelial cell culture. Our findings suggest that ANGPT1 increases pathophysiological angiogenesis in patients at risk of OHSS.

 
  
 

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Regulation of primordial follicle recruitment by cross-talk between the Notch and phosphatase and tensin homologue (PTEN)/AKT pathways 
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Lin-Qing Wang , Jing-Cai Liu , Chun-Lei Chen , Shun-Feng Cheng , Xiao-Feng Sun , Yong Zhao , Shen Yin , Zhu-Mei Hou , Bo Pan , Cheng Ding , Wei Shen and Xi-Feng Zhang
pp. 700-712

The growth of oocytes and the development of follicles require certain pathways involved in cell proliferation and survival. However, the mechanism of primordial follicle recruitment remains largely unknown. Inhibition of the Notch signalling pathway suppressed AKT phosphorylation and the proliferation of granulosa cells. Recruitment of primordial follicles was regulated by an interaction between the Notch and PI3K/AKT signalling pathways.

 
    | Supplementary Material (169 KB)
 

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Aberrant expression of TAR DNA binding protein-43 is associated with spermatogenic disorders in men 
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Divya Saro Varghese , Uma Chandran , Ambili Soumya , Sathy M. Pillai , Krishnapillai Jayakrishnan , Prabhakara P. Reddi and Pradeep G. Kumar
pp. 713-722

Defects in the molecular mechanisms involved in the production and normal functioning of spermatozoa could be a major cause of male factor infertility. We identified a strong association between the absence of a critical molecule in testicular germ cells and male factor infertility. Identification of such critical molecules and their subsequent use in diagnosing male factor infertility may improve infertility management strategies.

 
    | Supplementary Material (1 MB)
 

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N-Acetylcysteine improves oocyte and embryo quality in polycystic ovary syndrome patients undergoing intracytoplasmic sperm injection: an alternative to metformin 
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Ebrahim Cheraghi , Malek Soleimani Mehranjani , Mohammad Ali Shariatzadeh , Mohammad Hossein Nasr Esfahani and Zahra Ebrahimi
pp. 723-731

Although the number of oocytes in patients with polycystic ovary syndrome (PCOS) is usually greater than normal following ovulation stimulation, these oocytes are often of low quality. The aim of the present study was to investigate the effects of metformin and N-acetylcysteine (NAC) on follicular fluid parameters, as well as oocyte and embryo quality, in Iranian women with PCOS. We conclude that NAC improves oocyte and embryo quality, and could be administered as an alternative to metformin.

 
    | Supplementary Material (137 KB)
 

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Proteome profiling of the sperm maturation milieu in the rhesus monkey (Macaca mulatta) epididymis 
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Xin Liu , Shao-Hua Jin , Xue-Xia Liu , Wen-Juan Wang and Fu-Jun Liu
pp. 732-741

The first gel-based profiling of the rhesus monkey (Macaca mulatta) sperm-milieu proteome was established by a two-dimensional electrophoresis map. When compared with bull and human sperm-milieu proteins, those in the rhesus monkey had a close evolutionary relationship to human. It could be used as an ideal model for the development of diagnostic and therapeutic targets for infertility or contraception.

 
  
 

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CXADR-like membrane protein (CLMP) in the rat ovary: stimulation by human chorionic gonadotrophin during the periovulatory period 
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Feixue Li , Xiaoping Miao , Yonglong Chen and Thomas E. Curry
pp. 742-749

Cell–cell adhesion is important for cellular communication, cellular structure and steroidogenesis. The present study demonstrates that an ovulatory stimulus of human chorionic gonadotrophin (hCG) stimulates ovarian expression of the cell adhesion molecule CXADR-like membrane protein (CLMP). The hCG induction of Clmp mRNA is mediated through the protein kinase A signalling pathway by activation of the epidermal growth factor receptor and requires the synthesis of new protein. The temporal stimulation of ovarian CLMP may be important in cell adhesion events during ovulation and luteal formation.

 
  
 

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Influences of melatonin treatment, melatonin receptor 1A (MTNR1A) and kisspeptin (KiSS-1) gene polymorphisms on first conception in Sarda ewe lambs 
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S. Luridiana , M. C. Mura , C. Daga , G. Cosso , S. Bodano , F. Farci , F. Zidda and V. Carcangiu
pp. 750-756

Many factors are involved in the onset of reproductive activity in sheep, such as melatonin, its own receptor gene (MTNR1A) and kisspeptin (KiSS-1) gene. The aim of this study was to investigate if melatonin implants and MTNR1A and KiSS-1 gene polymorphisms can advance the first conception; we found that the G/G genotype of the MTNR1A gene improved the reproductive response to melatonin treatment. Understanding the mechanisms of melatonin function may improve reproductive performance in sheep.

 
  
 

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Neonatal testicular cell transplantation restores murine spermatogenesis damaged in the course of herpes simplex virus-induced orchitis 
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Ekaterina A. Malolina , Andrey Yu. Kulibin and Alla A. Kushch
pp. 757-764

There is growing evidence that viral infections of the genital tract may contribute to male infertility and cause orchitis. The present study was carried out to develop cell therapy for this disorder. We revealed that a combination of antiviral therapy and neonatal testis-cell transplantation is effective for restoration of murine spermatogenesis damaged by herpes simplex virus infection.

 
  
 

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Localisation and endocrine control of hyaluronan synthase (HAS) 2, HAS3 and CD44 expression in sheep granulosa cells 
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R. Chavoshinejad , W. F. A. Marei , G. M. Hartshorne and A. A. Fouladi-Nashta
pp. 765-775

We localised hyaluronan (HA) synthases (HAS) 1–3 and their main receptor CD44 in sheep antral follicles, showing intense staining in granulosa cells. Analyses of hormonal regulation of the HA system in cultured sheep granulosa cells indicated differential regulation of HAS2, HAS3 and CD44 expression by oestradiol, insulin-like growth factor, insulin and gonadotropins.

 
  
 

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Controlled hydrostatic pressure stress downregulates the expression of ribosomal genes in preimplantation embryos: a possible protection mechanism? 
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I. Bock , H. Raveh-Amit , E. Losonczi , A. C. Carstea , A. Feher , K. Mashayekhi , S. Matyas , A. Dinnyes and C. Pribenszky
pp. 776-784

Stress tolerance of gametes and embryos can be increased by preconditioning with hydrostatic pressure (HP) treatment. To study the effect of HP, mouse oocytes were treated, fertilised and then analysed by gene expression microarray at the 4-cell stage. The results revealed that numerous ribosomal genes were downregulated after HP treatment, suggesting that regulation of translation has a major role in hydrostatic pressure-induced stress tolerance.

 
    | Supplementary Material (15 KB)
 

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Association of C677T transition of the human methylenetetrahydrofolate reductase (MTHFR) gene with male infertility 
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Mohammad Karimian and Abasalt Hosseinzadeh Colagar
pp. 785-794

The human methylenetetrahydrofolate reductase (MTHFR) gene, located at chromosome 1p36.3, is associated with male infertility. Previous reports reported that the common polymorphisms C677T, A1298C and G1793A may be related to infertility. This study shows that the C677T transition plays an important role in the change in affinity of the MTHFR–Flavin adenine dinucleotide binding site, increasing the risk of infertility.

 
  
 

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Stable reference genes in granulosa cells of bovine dominant follicles during follicular growth, FSH stimulation and maternal aging 
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Muhammad Irfan-ur-Rehman Khan , Fernanda Caminha Faustino Dias , Isabelle Dufort , Vikram Misra , Marc-Andre Sirard and Jaswant Singh
pp. 795-805

Little is known about stability of reference genes during dominant follicle development. This study examined the expression of a number of granulosa cell genes during ovarian superstimulation and maternal aging in cattle. Results support the use of the geometric mean of four genes (UBE2D2, EIF2B2, GAPDH and SF3A1) for relative gene expression analyses of granulosa cells during maternal and follicular aging.

 
  
 

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Prepubertal male rats with high rates of germ-cell apoptosis present exacerbated rates of germ-cell apoptosis after serotonin depletion 
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Néstor Méndez Palacios , María Elena Ayala Escobar , Maximino Méndez Mendoza , Rubén Huerta Crispín , Octavio Guerrero Andrade , Javier Hernández Meléndez and Andrés Aragón Martínez
pp. 806-814

Genetic expression could be behind individual sensitivity to chemical substances. Here we show that individuals with innate low or high levels of germ-cell death have distinct gene expression and that levels of germ-cell death changed differentially after administration of amphetamine. This helps both to understand the exacerbated response to drugs by some individuals and to develop a rationale in individualised medicine.

 
    | Supplementary Material (22 KB)
 

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Sexual maturation of the Mongolian gerbil (Meriones unguiculatus): a histological, hormonal and spermatic evaluation 
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Maria Etelvina Pinto-Fochi , Ana Carolina Negrin , Wellerson Rodrigo Scarano , Sebastião Roberto Taboga and Rejane Maira Góes
pp. 815-823

Several reproductive aspects of Mongolian gerbil still remain unknown. This study determined the phases of sexual development of this rodent based on an integrative analysis of testicular morphology, hormonal data and sperm parameters. In terms of laboratory rodents, the gerbil has a similar reproductive profile with the rat. These reproductive data provide new foundations for future investigations involving toxicological or developmental studies with this species.

 
  
 

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Treatment of buffalo (Bubalus bubalis) donor cells with trichostatin A and 5-aza-2’-deoxycytidine alters their growth characteristics, gene expression and epigenetic status and improves the in vitro developmental competence, quality and epigenetic status of cloned embryos 
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M. Saini , N. L. Selokar , H. Agrawal , S. K. Singla , M. S. Chauhan , R. S. Manik and P. Palta
pp. 824-837

Skin cells are widely used as nuclear donors for producing cloned animals. We showed that treatment of these cells with certain chemical agents, which affect their DNA and the proteins associated with it, alters the characteristics and improves the clonability of these cells. This could help to improve the yield and quality of cloned embryos derived from skin cells.

 
    | Supplementary Material (323 KB)
 

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Cellular changes in the hamster testicular interstitium with ageing and after exposure to short photoperiod 
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E. Beltrán-Frutos , V. Seco-Rovira , C. Ferrer , J. F. Madrid , F. J. Sáez , M. Canteras and L. M. Pastor
pp. 838-851

The testicular interstitium is responsible for endocrine function in the testis. We have studied the cellular changes in two physiological processes of testicular involution, one reversible due to exposure to short photoperiod and the other irreversible due to ageing. These processes induce very different changes in the cells. These differences are probably related to cell turnover in the testicular interstitium and to the non-apoptotic but necrotic character of Leydig cell death.

 
  
 

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These articles have been peer reviewed and accepted for publication. They are still in production and have not been edited, so may differ from the final published form.

    RD15309  Accepted 25 April 2016
    Maternal food-restriction in rats of the F0 generation increases retroperitoneal fat and the number and size of adipocytes and induces periventricular astrogliosis in female F1 and male F2 generations.
    Andreia Joaquim, Cidéli Coelho, Priscila Motta, Luciano Felício, Eduardo Bondan, Elizabeth Teodorov, Maria Martins, Thiago Kirsten, Leoni Bonamin, Maria Bernardi
    Abstract


    RD16028  Accepted 23 April 2016
    Effects of intracerebroventricular infusions of ghrelin on secretion of follicle-stimulating hormone in peripubertal female sheep
    Anna Wójcik-GÅ‚adysz, Marta WaÅ„kowska, Alina Gajewska, Tomasz Misztal, Marlena ZieliÅ„ska-Górska, MichaÅ‚ Szlis, Jolanta Polkowska
    Abstract


    RD16145  Accepted 20 April 2016
    The role of Ca2+ in the in vitro maturation of the sterlet Acipenser ruthenus spermatozoa
    Olga Bondarenko, Boris Dzyuba, Marek Rodina, Jacky Cosson
    Abstract


    RD16033  Accepted 20 April 2016
    Actions of activin A, connective tissue growth factor, hepatocyte growth factor and teratocarcinoma-derived growth factor 1 on development of the bovine preimplantation embryo
    Jasmine Kannampuzha-Francis , Paula Tribulo, Peter Hansen
    Abstract


    RD15391  Accepted 20 April 2016
    Iloprost supports early development of in vitro-produced porcine embryos through activation of the PI3K/AKT signaling pathway
    Pil-Soo Jeong, Seung-Bin Yoon, Seon-A Choi, Bong-Seok Song, Ji-Su Kim, Bo-Woong Sim, Young-Ho Park, Hae-Jun Yang, Seong-Eun Mun, Young-hyun Kim, Philyong Kang, Kang-Jin Jeong, Youngjeon Lee, Young Bae Jin, Jae-Won Huh, Sang-Rae Lee, Deog-Bon Koo, Young Il Park, Sun-Uk Kim, Kyu-Tae Chang
    Abstract


    RD15483  Accepted 07 April 2016
    Individual differences in the distribution of acrosomal tyrosine-phosphorylated proteins in epididymal and ejaculated spermatozoa of Japanese Black bulls
    Miyuki Arai, Kenta Minami, Yukari Ogura, Nagisa Otsuka, Shohei Hama, Hiroshi Harayama, Mitsuhiro Sakase, Moriyuki Fukushima
    Abstract


    RD16026  Accepted 06 April 2016
    Pak1 activity is required for H3 Ser10 phosphorylation and chromatin condensation in mouse oocyte meiosis
    Nana Zhang, Xiuhong Li, Xiaoyun Liu, Yan Cao, Dandan Chen, Xiaoyu Liu, Qian Wang, Juan Du, Jing Weng, Wei Ma
    Abstract


    RD15538  Accepted 04 April 2016
    Expressions of lipoprotein receptors and cholesterol efflux regulatory proteins during luteolysis in bovine corpus luteum
    Kei Horihata, Shin Yoshioka, Masahiro Sano, Yuki Yamamoto, Koji Kimura, Dariusz Skarzynski, Kiyoshi Okuda
    Abstract


    RD15526  Accepted 05 April 2016
    Potential bleaching effects on coral reproduction
    Mary Hagedorn, Virginia Carter, Claire Lager, Julio Camperio Ciani, Alison Dygert, Reuben Schleiger , E. Henley
    Abstract


    RD15253  Accepted 31 March 2016
    Variable duration of reproductive suppression in male coyotes (Canis latrans) treated with a high dose of the GnRH agonist deslorelin
    Marjorie MacGregor, Cheryl Asa, Donal Skinner
    Abstract


    RD16077  Accepted 31 March 2016
    Aquaglyceroporins 3 and 7 in bull sperm: identification, localisation and their relationship with sperm cryotolerance
    Noelia Prieto-Martínez, Roser Morató, Rodrigo Muiño, Carlos O. Hidalgo, Joan Rodriguez-Gil, Sergi Bonet, Marc Yeste
    Abstract


    RD15330  Accepted 28 March 2016
    The effects of sildenafil citrate on the fetoplacental development and hemodynamics in a rabbit model of intrauterine growth restriction
    Jorge Lopez-Tello, Maria Arias-Alvarez, Maria-Ángeles Jiménez-Martínez, Alicia Barbero, Rosa Maria García-García, Maria Rodriguez, Pedro Lorenzo, Laura Torres-Rovira, Susana Astiz, Antonio Gonzalez-Bulnes, Pilar Rebollar
    Abstract


    RD15357  Accepted 26 March 2016
    Non-genomic action of Vitamin D3 on NMDA and kainate receptor-mediated actions in juvenile GnRH neurons
    Pravin Bhattarai, Janardhan Bhattarai, Min Sun Kim, Seong Kyu Han
    Abstract


    RD15414  Accepted 24 March 2016
    Membrane lipid profile of in vitro-produced embryos is impacted by vitrification but not by long-term dietary supplementation of polyunsaturated fatty acids for oocyte donor beef heifers
    Beatriz Leão, Nathalia Rocha, Eriklis Nogueira, Elaine Cabral, Christina Ferreira, Marcos Eberlin, Monica Accorsi, Thiago Neves, Gisele Mingoti
    Abstract


    RD15209  Accepted 25 March 2016
    Roles of interferon-stimulated gene 15-kDa protein in bovine embryo development
    Shuan Zhao, Yi Wu, Hui Gao, Alexander Evans, Shenming Zeng
    Abstract


    RD15432  Accepted 17 March 2016
    Prominin-1 glycosylation changes throughout early pregnancy in uterine epithelial cells under the influence of maternal ovarian hormones
    Samson Dowland, Romanthi Madawala, Connie Poon, Laura Lindsay, Chris Murphy
    Abstract


    RD15415  Accepted 17 March 2016
    Temporal regulation of ERK-1/2 phosphorylation and HSP70 and ATF3 during PGF-induced luteal regression in pseudopregnant rats following heat stress
    Wujiao Bai, Pengjin Jin, Meiqian Kuang, Quanwei Wei, Fangxiong Shi, Davis John, Dagan Mao
    Abstract


    RD16090  Accepted 16 March 2016
    The effects of TNF-α and its antagonist “Etanercept” in ejaculated human sperm: an in vitro study
    Nicola Pascarelli, Antonella Fioravanti, Elena Moretti, Giacomo Guidelli, Lucia Mazzi, Giulia Collodel
    Abstract


    RD16020  Accepted 17 March 2016
    Implantation rates subsequent to the transfer of embryos produced in different phases during double stimulation of poor ovarian responder: a retrospective study
    Qiong Zhang, Xiaoyue Guo, Yanping Li
    Abstract


    RD15385  Accepted 15 March 2016
    Effects of foetal genotype and sex on developmental response to maternal malnutrition
    Laura Cogollos, Consolacion Garcia-Contreras, Marta Vazquez-Gomez, Susana Astiz, Raul Sanchez-Sanchez, Ernesto Gomez-Fidalgo, Cristina Ovilo, Beatriz Isabel, Antonio Gonzalez-Bulnes
    Abstract


    RD15290  Accepted 13 March 2016
    Anti-Müllerian hormone (AMH) reduces growth rate without altering follicular survival in isolated caprine preantral follicles cultured in vitro
    Rebeca Rocha, Laritza Lima, Ivina Brito, Gerlane Silva, Hudson Correia, Naiza Sá, Anna Ferreira, Antonia Débora Sales, Carlos Lobo, Claudio Campello, Johan Smitz, Matthew Wheeler, José Figueiredo
    Abstract


    RD15517  Accepted 12 March 2016
    Perinatal exposure to insecticide fipronil: effects on reproductive system in male rats.
    Aline de Barros, Julie Bae, Cibele Borges, Josiane Rosa, Marilia Cavariani, Patrícia Silva, Patricia Pinheiro, Janete Anselmo-Franci, Arielle Arena
    Abstract


    RD15492  Accepted 09 March 2016
    Progesterone alters phosphorylation of motility associated proteins in human spermatozoa
    Vrushali Sagare-Patil, Deepak Modi
    Abstract


    RD15494  Accepted 19 February 2016
    Optineurin suppression activates the mediators involved in the terminal effector pathways of human labour and delivery
    Ratana Lim, Gillian Barker, Martha Lappas
    Abstract


    RD15521  Accepted 11 February 2016
    Cerium oxide nanoparticles (CeO2 NPs) improve the developmental competence of in vitro matured prepubertal ovine oocytes
    Federica Ariu, Luisa Bogliolo, Alessandra Pinna, Luca Malfatti, Plinio Innocenzi, Laura Falchi, Daniela Bebbere, Sergio Ledda
    Abstract


    RD15412  Accepted 02 February 2016
    Saving Wild Ungulate Diversity Through Enhanced Management and Sperm Cryopreservation
    Budhan Pukazhenthi
    Abstract


    RD15261  Accepted 05 January 2016
    Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates
    Claire Kershaw, Gareth Evans, Rachael Rodney, Chis Maxwell
    Abstract


    RD15079  Accepted 03 March 2015
    A developmental tale - metabolism takes centre stage
    Roger Sturmey, Rebecca Krisher
    Abstract


    RD14181  Accepted 19 January 2015
    Time within reproductive season, but not age or inbreeding coefficient, influences seminal and sperm quality in the whooping crane (Grus americana)
    Megan Brown, Sarah Converse, Jane Chandler, Adrienne Crosier, Warren Lynch, David Wildt, Carol Keefer, Nucharin Songsasen
    Abstract


    RD12339  Accepted 19 October 2012
    The critical importance of ovarian angiogenesis
    Bob Robinson
    Abstract


30


The Most Read ranking is based on the number of downloads from the CSIRO PUBLISHING website of articles published in the previous 12 months. Usage statistics are updated daily.

Rank Paper Details
1. Published 17 April 2015
Nutrition and maternal metabolic health in relation to oocyte and embryo quality: critical views on what we learned from the dairy cow model

Jo L. M. R. Leroy, Sara D. M. Valckx, Lies Jordaens, Jessie De Bie, Karolien L. J. Desmet, Veerle Van Hoeck, Jack H. Britt, Waleed F. Marei and Peter E. J. Bols

2. Published 8 July 2015
Maternal age and in vitro culture affect mitochondrial number and function in equine oocytes and embryos

W. Karin Hendriks, Silvia Colleoni, Cesare Galli, Damien B. B. P. Paris, Ben Colenbrander, Bernard A. J. Roelen and Tom A. E. Stout

3. Published 17 April 2015
Blastocyst metabolism

David K. Gardner and Alexandra J. Harvey

4. Published 17 April 2015
History of oocyte and embryo metabolism

Henry J. Leese

5. Published 3 June 2015
DNA methylation reprogramming during oogenesis and interference by reproductive technologies: Studies in mouse and bovine models

Ellen Anckaert and Trudee Fair

6. Published 3 December 2015
IETS 2016 abstracts


7. Influence of bovine serum albumin and fetal bovine serum supplementation during in vitro maturation on lipid and mitochondrial behaviour in oocytes and lipid accumulation in bovine embryos

Maite del Collado, Naiara Z. Saraiva, Flavia L. Lopes, Roberta C. Gaspar, Luciana C. Padilha, Roberta R. Costa, Guilherme F. Rossi, Roberta Vantini and Joaquim M. Garcia

8. Published 8 July 2015
Maternal control of early embryogenesis in mammals

Kun Zhang and George W. Smith

9. Published 8 July 2015
Antioxidant supplementation during in vitro culture improves mitochondrial function and development of embryos from aged female mice

Elena Silva, Alison F. Greene, Kevin Strauss, Jason R. Herrick, William B. Schoolcraft and Rebecca L. Krisher

10. Published 17 April 2015
Hyperglycaemia and lipid differentially impair mouse oocyte developmental competence

Siew L. Wong, Linda L. Wu, Rebecca L. Robker, Jeremy G. Thompson and Melanie L. Sutton McDowall

11. Published 17 April 2015
Nutrient pathways regulating the nuclear maturation of mammalian oocytes

Stephen M. Downs

12. Published 8 July 2015
Effects of age on follicular fluid exosomal microRNAs and granulosa cell transforming growth factor-β signalling during follicle development in the mare

Juliano C. da Silveira, Quinton A. Winger, Gerrit J. Bouma and Elaine M. Carnevale

13. Published 17 April 2015
Adverse effects of obesity and/or high-fat diet on oocyte quality and metabolism are not reversible with resumption of regular diet in mice

Kasey A. Reynolds, Anna L. Boudoures, Maggie M.-Y. Chi, Qiang Wang and Kelle H. Moley

14. Published 8 July 2015
Not just a number: effect of age on fertility, pregnancy and offspring vigour in thoroughbred brood-mares

Charles F. Scoggin

15. Published 17 April 2015
Do little embryos make big decisions? How maternal dietary protein restriction can permanently change an embryo’s potential, affecting adult health

Tom P. Fleming, Adam J. Watkins, Congshan Sun, Miguel A. Velazquez, Neil R. Smyth and Judith J. Eckert

16. Published 8 July 2015
Oogonial stem cells as a model to study age-associated infertility in women

Neha Garg and David A. Sinclair

17. Published 8 July 2015
Centrosome and microtubule functions and dysfunctions in meiosis: implications for age-related infertility and developmental disorders

Heide Schatten and Qing-Yuan Sun

18. Published 17 April 2015
One-carbon metabolism and epigenetic regulation of embryo development

Juan Xu and Kevin D. Sinclair

19. Published 17 April 2015
Ambient ionisation mass spectrometry for lipid profiling and structural analysis of mammalian oocytes, preimplantation embryos and stem cells

Christina R. Ferreira, Alan K. Jarmusch, Valentina Pirro, Clint M. Alfaro, Andres F. González-Serrano, Heiner Niemann, Matthew B. Wheeler, Rathnaweera A. C. Rabel, Judy E. Hallett, Rebecca Houser, Annemarie Kaufman and R. Graham Cooks

20. Published 17 April 2015
Applying metabolomic analyses to the practice of embryology: physiology, development and assisted reproductive technology

Rebecca L. Krisher, Adam L. Heuberger, Melissa Paczkowski, John Stevens, Courtney Pospisil, Randall S. Prather, Roger G. Sturmey, Jason R. Herrick and William B. Schoolcraft


      
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