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Reproduction, Fertility and Development
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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: Tony Flint 

 
 
 

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Published online 30 September 2014
Changes in myometrial expression of progesterone receptor membrane components 1 and 2 are associated with human parturition at term 
Ray Wang, Penelope M. Sheehan and Shaun P. Brennecke

The progesterone receptor membrane components 1 and 2 (PGRMC1, PGRMC2) may be a part of the mechanism by means of which women come into labour despite continued high progesterone concentrations. Expression of PGRMC1 mRNA and protein was found to be significantly lower in myometrium from women in labour. This change may be mediated by microRNA species hsa-miR-520e.

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Published online 25 September 2014
Ultrastructure and mitochondrial numbers in pre- and postpubertal pig oocytes 
Hanne Skovsgaard Pedersen, Henrik Callesen, Peter Løvendahl, Fenghua Chen, Jens Randel Nyengaard, Nanett Kvist Nikolaisen, Peter Holm and Poul Hyttel

Prepubertal pig oocytes, used for the production of transgenic pigs as human disease models, have low developmental competence. The aim of this experiment was to describe oocyte ultrastructure and to quantify mitochondria in pre- and postpubertal pig oocytes. The ultrastructure and higher number of mitochondria in large pre- and postpubertal oocytes was compatible with higher developmental competence.

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Published online 24 September 2014
Oncostatin M and leukaemia inhibitory factor trigger signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 pathways but result in heterogeneous cellular responses in trophoblast cells 
Wittaya Chaiwangyen, Stephanie Ospina-Prieto, Diana M. Morales-Prieto, Francisco Lazaro Pereira de Sousa, Jana Pastuschek, Justine S. Fitzgerald, Ekkehard Schleussner and Udo R. Markert

Leukemia inhibitory factor (LIF) and Oncostatin M (OSM) play important roles during implantation and placentation. This study aims to compare responses on LIF and OSM in four different trophoblastic cell lines. In all, LIF and OSM induce STAT3 and ERK1/2 phosphorylation, while only LIF significantly increases trophoblast cell invasion. Thus, both factors have similar but not identical funtions in placentation.

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Published online 19 September 2014
Decreased expression of aquaporin 2 is associated with impaired endometrial receptivity in controlled ovarian stimulation 
Dan Zhang, Gufeng Xu, Runju Zhang, Yimin Zhu, Huijuan Gao, Caiyun Zhou, Jianzhong Sheng and Hefeng Huang

This study revealed the role of auquaporin 2 (AQP2, a water channel) in impaired embryo implantation following ovarian stimulation. It was found that AQP2 was decreased in endometria of ovarian stimulated women and suppressing AQP2 led to impaired implantation. The study indicated the importance of water transportation in embryo implantation and suggested a new way to improve outcome of assisted conception.

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Published online 12 September 2014
Decreased melatonin levels and increased levels of advanced oxidation protein products in the seminal plasma are related to male infertility 
Ewa Maria Kratz, Agnieszka Piwowar, Michal Zeman, Katarína Stebelová and Theresia Thalhammer

Melatonin is a wide-spectrum antioxidant; therefore, the aim of this study was to correlate advanced oxidation protein products (AOPPs) and the total antioxidant capacity (TAC) with melatonin levels in the seminal plasma of men with azoospermia, theratozoospermia and fertile controls. Decreased levels of melatonin and AOPP altered the oxidative–antioxidative balance in the ejaculate, thereby reducing fertility. These parameters may serve as additional markers of semen quality and male reproductive potential.

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Published online 11 September 2014
Discovery of human posterior head 20 (hPH20) and homo sapiens sperm acrosome associated 1 (hSPACA1) immunocontraceptive epitopes and their effects on fertility in male and female mice 
Xuemei Chen, Xiaodong Liu, Xiuhua Ren, Xuewu Li, Li Wang and Weidong Zang

Low antibody titres and the irreversible suppression of spermatogenesis are current hurdles in the contraceptive vaccine field. The purpose of immunocontraception research is to obtain prominent and reversible contraceptive effect through vaccines administered to both males and females. These peptides could be used in the specific diagnosis and treatment of male and female infertility as well as contraceptive vaccine development.

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Published online 11 September 2014
Why we should not select the faster embryo: lessons from mice and cattle 
Alfonso Gutierrez-Adan, Carlee R. White, Ann Van Soom and Mellissa R. W. Mann

Many studies have shown that in vitro culture can negatively impact preimplantation development, which necessitates some selection criteria to identify the best-suited embryos for transfer. With emerging data in bovine and mouse supporting increased developmental competency in embryos with moderate rates of development, it is time to question whether the fastest developing embryos are the best embryos for transfer in the human clinic. In this review we explore evidence surrounding the question of developmental timing as it relates to bovine embryo quality, mouse embryo quality and genomic imprint maintenance, and embryo sex.

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Published online 11 September 2014
The effects of permeating cryoprotectants on intracellular free-calcium concentrations and developmental potential of in vitro-matured feline oocytes 
Jason R. Herrick, Chunmin Wang and Zoltan Machaty

Changes in intracellular concentrations of Ca2+, hardening of the zona pellucida, parthenogenetic activation and embryonic development following IVF were evaluated in feline oocytes after exposure to three common cryoprotectants.  A combination of DMSO and propanediol had minimal effects on the viability of feline oocytes and may be useful for the development of more efficient protocols for vitrification of feline oocytes.

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Published online 11 September 2014
Effects of whole-body heat on male germ cell development and sperm motility in the laboratory mouse 
H. Wechalekar, B. P. Setchell, K. R. Pilkington, C. Leigh, W. G. Breed and E. Peirce

Putative effects of whole-body heat exposure include changes in testicular cells and spermatozoa. This study compared laboratory mice exposed to whole-body heat to control mice. The results show reduced testis weights and damage to developing and formed spermatozoa. These changes can cause male infertility. The results from this study could be extrapolated to humans exposed to high temperatures in their occupations.

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Published online 09 September 2014
Epigenetic processes in the male germline 
Alan M. O'Doherty and Paul A. McGettigan

Epigenetic modification of the mammalian genome is indispensable for successful development.  Many of these modifications, such as DNA methylation, are established and removed during gametogenesis, following fertilisation and during the early stages of embryogenesis.  These epigenetic modifications occur at different times during meiosis, depending on whether they occur during spermatogenesis or oogenesis.  Similarly, post-fertilisation modification dynamics occur differently depending on the parental origin of the genomic material.  This review will focus on these processes in the male germline.

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Published online 05 September 2014
Role of Wnt signalling in early pregnancy 
Sarmah B. Nayeem, Frank Arfuso, Arun Dharmarajan and Jeffrey A. Keelan

The Wnt family of secreted ligands plays a major role in regulating a wide range of biological processes, including reproduction. This review summarises our present knowledge of the expression, regulation and function of Wnt signalling pathways in early pregnancy and placentation. Knowledge in this area has applications in the diagnosis, prevention and treatment of infertility and gestational diseases.

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Published online 02 September 2014
Prolactin modulates luteal regression from the coeliac ganglion via the superior ovarian nerve in the late-pregnant rat 
Sandra S. Vallcaneras, Magalí de la Vega, Silvia M. Delgado, Alicia Motta, Carlos Telleria, Ana M. Rastrilla and Marilina Casais

This work provides evidence of the functional interaction between the endocrine and peripheral neural systems. The aim was to study the effects of prolactin added to the coeliac ganglion, via the superior ovarian nerve, on ovarian physiology at the end of rat pregnancy. Prolactin acts at the coeliac ganglion and sends information to the ovary by a neural pathway, modifying parameters of functional and structural luteal regression.

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Published online 28 August 2014
Stress preconditioning of semen before cryopreservation improves fertility and increases the number of offspring born: a prospective randomised study using a porcine model 
A. Horváth, O. Szenci, K. Nagy, L. Végh and Cs. Pribenszky

Standard cryopreservation protocols and modified AI techniques are required to achieve acceptable fertility results with frozen–thawed boar semen. Using simplified procedures, but applying a sublethal hydrostatic pressure to the semen at room temperature before cryopreservation, yielded higher motility and fertility results. The AI dosage still needs to be optimised and, because no direct correlation was found between motility and fertility on an individual basis, the motility results cannot be used to predict potential fertility.

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Published online 28 August 2014
Seminal plasma affects sperm sex sorting in boars 
Diego V. Alkmin, Inmaculada Parrilla, Tatiana Tarantini, David del Olmo, Juan M. Vazquez, Emilio A. Martinez and Jordi Roca

Sex-sorted spermatozoa can play a crucial role in enhancing genetic improvement and production efficiency in swine industry. The presence of a high proportion of seminal plasma in semen samples negatively influences sperm sex-sorting efficiency. Removing the seminal plasma before sorting can reverse the consistent poor sperm sortability exhibited by the ejaculates of some boars.

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Published online 28 August 2014
Human extravillous trophoblast invasion: intrinsic and extrinsic regulation 
E. Menkhorst, A. Winship, M. Van Sinderen and E. Dimitriadis

Invasion of specialised fetal cells, the extravillous trophoblast, into the uterine decidua is critical for the establishment of pregnancy. Overall, there exists an exquisite balance between trophoblast over- and under-invasion. This review summarises what is known about the intrinsic invasive potential of the extravillous trophoblast and the role of maternal cells in regulating invasion. A greater understanding of the mechanisms will lead to therapeutics to treat at-risk pregnancies.

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Published online 26 August 2014
Signalling pathways involved in the synergistic effects of human growth differentiation factor 9 and bone morphogenetic protein 15 
Karen L. Reader, David G. Mottershead, Georgia A. Martin, Robert B. Gilchrist, Derek A. Heath, Kenneth P. McNatty and Jennifer L. Juengel

The oocyte-derived growth factors, bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), play important roles in fertility, but their mechanism of action differs between species. Our aim was to determine the signalling pathways used by human BMP15 and GDF9; we found that they act synergistically and the combinations of pathways used are different from those observed in other species. Understanding the mechanisms of function of these human growth factors may help tailor treatments to improve fertility.

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Published online 25 August 2014
Interleukin 11: similar or opposite roles in female reproduction and reproductive cancer? 
Amy Winship, Ellen Menkhorst, Michelle Van Sinderen and Evdokia Dimitriadis

A signalling protein important in pregnancy can also promote uterine tumour growth in women. High levels of interleukin (IL)-11 protein are found in the placenta of pregnant women who have developed the potentially life-threatening condition of pre-eclampsia. The same protein is also highly expressed in tumours that develop in the endometrium, the inner lining of the uterus. IL-11 may be a new potential therapeutic target for cancer and the pregnancy complication pre-eclampsia.

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Published online 22 August 2014
Oxygen modulates human embryonic stem cell metabolism in the absence of changes in self-renewal 
Alexandra J. Harvey, Joy Rathjen, Lijia Jackie Yu and David K. Gardner

In this study we characterise the ability of environmental oxygen to regulate human embryonic stem cells (hESC). We show that the oxygen concentration influences metabolite use, metabolic gene expression, cell death and free radical production in hESC, without affecting the expression of characteristic hESC markers. Our findings suggest that hESC benefit from culture in a reduced oxygen environment that mimics the physiological paradigm.

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Published online 22 August 2014
Role of linker histone H1c during the reprogramming of Chinese swamp buffalo (Bubalus Bubalis) embryos produced by somatic cell nuclear transfer 
Gao-Bo Huang, Li Quan, Yong-Lian Zeng, Jian Yang, Ke-Huan Lu and Sheng-Sheng Lu

The movement of numerous proteins between the cytoplasm and nucleus is essential to remodelling. In this study, the exchange of histone between the cytoplasm of the oocyte (H1foo) and the transferred nucleus (H1c) is shown to be essential to somatic cell nuclear transfer in buffalo.

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Published online 20 August 2014
Variation in seminal plasma alters the ability of ram spermatozoa to survive cryopreservation 
J. P. Rickard, R. E. Schmidt, J. W. Maddison, R. Bathgate, G. W. Lynch, X. Druart and S. P. de Graaf

The effect of seminal plasma on sperm function is yet to be fully explained. This study demonstrates that survival of spermatozoa during cryopreservation is modified by the source of seminal plasma (high or low freezing resilient male) to which they are exposed prior to freezing. These results demonstrate that seminal plasma varies in composition and is the first step in identifying its positive and negative components.

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Published online 13 August 2014
Analysis of STAT1 expression and biological activity reveals interferon-tau-dependent STAT1-regulated SOCS genes in the bovine endometrium 
A. Vitorino Carvalho, C. Eozenou, G. D. Healey, N. Forde, P. Reinaud, M. Chebrout, L. Gall, N. Rodde, A. Lesage Padilla, C. Giraud Delville, M. Leveugle, C. Richard, I. M. Sheldon, P. Lonergan, G. Jolivet and O. Sandra

Implantation is a critical step of mammalian pregnancy whose progression depends on the quality of embryo and female genital tract. Using cattle as an animal model, gene expression, regulation and cell localization of STAT1 transcription factor were investigated in the endometrium during the oestrous cycle and early pregnancy; direct STAT1 target genes were identified by chromatin immunoprecipitation run with primary cultures of bovine endometrial cells. Altogether our data provide new insights on STAT1 biological functions in the endometrium during pregnancy maternal recognition and implantation.

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Published online 13 August 2014
Low physiological levels of prostaglandins E2 and F improve human sperm functions 
Mariana Rios, Daniela V. Carreño, Carolina Oses, Nelson Barrera, Bredford Kerr and Manuel Villalón

The recent discovery that prostaglandins are present in the fluid of the female genital tract and increase the motility of human sperm in vitro could lead to a possible treatment to improve male infertility. The findings of this study that incubation of human spermatozoa with low physiological levels of PGE2 or PGF increases sperm functions could improve conditions for assisted reproduction protocols and help infertile couples seeking medical help, which, according to global estimates, could be as many as 50–80 million people.

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Published online 13 August 2014
The requirement for protein kinase C delta (PRKCD) during preimplantation bovine embryo development 
Qi-En Yang, Manabu Ozawa, Kun Zhang, Sally E. Johnson and Alan D. Ealy

Embryonic loss is one of the major contributors to infertility in humans and domestic animals. This work determined that protein kinase C delta, an intracellular signalling molecule, is involved in several important developmental processes during early embryonic development in cattle. These outcomes implicate this molecule as a principal player in establishing the developmental potential of bovine embryos.

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Published online 11 August 2014
A new technique for analysis of human sperm morphology in unstained cells from raw semen 
Carles Soler, Almudena García-Molina, María Sancho, Jesús Contell, Manuel Núñez and Trevor G. Cooper

We describe the first technique that makes it possible to analyse the real size and shape of sperm cells. The development and application of this technique must lead to a new classification of human sperm morphology. This new approach will provide a revolutionary tool in the definition of human male fertility.

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Published online 11 August 2014
Yolk size and ovulation order determine fertility within double-yolked duck (Anas platyrhynchos domesticus) eggs 
Attila Salamon and John P. Kent

In double-yolked duck eggs, the first yolk ovulated achieved a higher level of fertilisation (68.82%) than the second yolk (34.98%), demonstrating a primacy effect. Eggs with larger yolks achieved a higher level of fertilisation, demonstrating an optimal yolk size effect. Both primacy and optimal size have a positive effect on the successful fertilisation of eggs in species with internal fertilisation.

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Published online 01 August 2014
Effects of maternal undernutrition during late gestation and/or lactation on colostrum synthesis and immunological parameters in the offspring 
S. Chadio, A. Katsafadou, B. Kotsampasi, G. Michailidis, K. C. Mountzouris, D. Kalogiannis and V. Christodoulou

Colostrum, apart from its crucial role for neonate survival, serves as a conduit for the transmission of signalling molecules from the mother to offspring. Maternal nutrition during late gestation affects colostrum composition and immune factor content, which through immunomodulatory properties, alters immune system function in the offspring. Events during fetal or neonatal life may have programming effects on immune function in later life.

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Published online 28 July 2014
Expression of group III metabotropic glutamate receptors in the reproductive system of male mice 
Marcin Marciniak, Barbara Chruścicka, Tomasz Lech, Grzegorz Burnat and Andrzej Pilc

Although glutamate signalling is traditionally investigated in the context of the central nervous system, it could also play a role in non-neuronal tissues. Herein, we investigated the expression of group III metabotropic glutamate (mGlu) receptors in the reproductive system of male mice. Based on findings for the mGlu7 receptor, we postulate that constitutively expressed mGlu receptors in the male reproductive system may influence male fertility.

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Published online 24 July 2014
Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Müllerian hormone profiles 
G. Gamarra, C. Ponsart, S. Lacaze, B. Le Guienne, P. Humblot, M.-C. Deloche, D. Monniaux and A. A. Ponter

Ovum pick-up and in vitro embryo production is a technique that can be used in high genetic merit dairy heifers to produce large numbers of high quality embryos. Serum anti-Müllerian hormone (AMH) was positively correlated with the numbers and quality of in vitro produced embryos after superovulation. A simple dietary supplement, propylene glycol, which increased circulating insulin, glucose and IGF1, was able to further increase oocyte quality and embryo production in animals with high AMH levels.

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Published online 24 July 2014
Variations in epithelial Na+ transport and epithelial sodium channel localisation in the vaginal cul-de-sac of the brushtail possum, Trichosurus vulpecula, during the oestrous cycle 
T.-A. Alsop, B. J. McLeod and A. G. Butt

The vaginal cul-de-sac, which is unique to the reproductive tract of marsupials, is thought to play an important role in reproduction, as the volume of fluid in the cul-de-sac increases up to ovulation and diminishes thereafter. Generally, changes in fluid volume in the reproductive tract are a result of changes in the transport properties of the epithelial lining. Our findings describe an epithelial transport mechanism responsible for absorption of fluid from the cul-de-sac following ovulation.

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Published online 22 July 2014
N-Glycans in Xenopus laevis testis characterised by lectin histochemistry 
Galder Valbuena, Juan Francisco Madrid, María Martínez de Ubago, Laura Gómez-Santos, Edurne Alonso, Lucio Díaz-Flores and Francisco J. Sáez

N-glycans (oligosaccharide chains linked to proteins by the amino group of an asparagine) essentials for mammalian spermatogenesis have been identified. Here, for the first time the presence of N-glycans in the testis of the amphibian Xenopus laevis has been analysed by lectin histochemistry. The functional role of these glycans could be related to the role of N-glycans involved in mammalian spermatogenesis.

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Published online 18 July 2014
Effect of combined exogenous progesterone with luteotrophic support via equine chorionic gonadotrophin (eCG) on corpus luteum development, circulating progesterone concentrations and embryo development in cattle 
L. O'Hara, N. Forde, P. Duffy, F. Randi, A. K. Kelly, A. Valenza, P. Rodriguez and P. Lonergan

Low circulating progesterone in the week following ovulation has been implicated in poor fertility in cattle. The aim of this study was to examine the effect of exogenous progesterone in combination with a luteotrophic agent on corpus luteum survival and conceptus development in cross-bred beef heifers. Results may provide the basis for a strategy to elevate progesterone and improve fertility.

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Published online 18 July 2014
Superovulation alters embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) gene expression in mouse oocytes and early embryos 
Saffet Ozturk, Aylin Yaba-Ucar, Berna Sozen, Derya Mutlu and Necdet Demir

Superovulation adversely influences oocyte maturation and early embryo development, but how superovulation results in these disturbances has not been clearly addressed yet. In this study, we aimed to determine the effect of superovulation on expression of embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) genes. We found that superovulation significantly alters Epab and Pabpc1 gene expression in mouse oocytes and early embryos.

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Published online 18 July 2014
In vitro interaction between resistin and peroxisome proliferator-activated receptor γ in porcine ovarian follicles 
Agnieszka Rak-Mardy?a and Eliza Drwal

Resistin is a small cysteine-rich protein, produced by adipose tissue and involved in insulin resistance, type 2 diabetes and reproduction. The findings of this study suggest that there is local cooperation exists resistin and PPARγ expression in the porcine ovary; resistin significantly increases PPARγ expression, whereas PPARγ decreases resistin expression. PPARγ is a new key regulator of resistin expression and function.

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Published online 18 July 2014
Expression patterns of mitochondrial OXPHOS components, mitofusin 1 and dynamin-related protein 1 are associated with human embryo fragmentation 
Vesna Otasevic, Lela Surlan, Milica Vucetic, Ivan Tulic, Biljana Buzadzic, Ana Stancic, Aleksandra Jankovic, Ksenija Velickovic, Igor Golic, Milica Markelic, Aleksandra Korac and Bato Korac

Improving the quality of preimplantation embryos is imperative in human in vitro fertilisation since the birth of the first baby in 1978, and high fragmentation is one of the limiting factors. This study links the phenomenon of human embryo fragmentation with energy homeostasis, mitochondrial dynamics and oxidative phosphorylation, suggesting altered transcriptional profiles of mitochondria-encoded components. A better understanding of the molecular mechanisms underlying these processes may help improve the energy capacity of the early embryo.

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Published online 17 July 2014
Mitochondrial activity during pre-maturational culture in in vitro-grown bovine oocytes is related to maturational and developmental competences 
Weiping Huang, Sung-Sik Kang, Katsuhisa Nagai, Yojiro Yanagawa, Yoshiyuki Takahashi and Masashi Nagano

Clarifying the mechanism for acquisition of developmental competence in bovine oocytes is important for improving the in vitro production system. Relationships between developmental competence and mitochondrial activity of in vitro-grown oocytes during meiotic arrest immediately before IVM culture were investigated. Mitochondrial activity at 10 h after starting meiotic arrest was highest and those oocytes showed comparable developmental competence with in vivo-grown oocytes.

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Published online 16 July 2014
Efficient generation of sFat-1 transgenic rabbits rich in n-3 polyunsaturated fatty acids by intracytoplasmic sperm injection 
Shun Zhang, Fenghua Lu, Qingyou Liu, Yubing Liu, Xiaomei Guan, Yingming Wei, Shijian Tan and Deshun Shi

N-3 PUFAs are beneficial to human health, and increasing the amount of n-3 PUFAs in mammals by introducing sFat-1 gene will improve the animal meat quality. This study was undertaken to produce sFat-1 transgenic rabbit by ICSI, and found that sFat-1 transgenic rabbits exhibited an ~10-fold increase in n-3 PUFAs in muscles. It will be possible to produce a new transgenic rabbit breed with high n-3 PUFAs.

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Published online 15 July 2014
Low-density lipoprotein receptor affects the fertility of female mice 
Tao Guo, Liang Zhang, Dong Cheng, Tao Liu, Liguo An, Wei-Ping Li and Cong Zhang

Low density lipoprotein receptor (LDLR) is responsible for the uptake of cholesterol-carrying lipoprotein particles into cells. The Ldlr deficient mice developed severe dyslipidemia and had significantly decreased female fertility. This study provides additional evidence for the role of Ldlr in regulation of lipid metabolism as well as female reproductive health during postnatal life.

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Published online 15 July 2014
Low level of basal testosterone: a significant risk factor for poor oocyte yield after ovulation induction 
Jing Guo, Qingxue Zhang, Yu Li, Wenjun Wang and Dongzi Yang

This study supplies a new predictor for poor ovarian response (POR) in the in vitro fertilisation field. We investigated the association of low basal androgen levels and IVF outcomes in a large Chinese population, and found low basal testosterone (<0.88 nmol L–1) was an independent risk factor for POR. This finding may help to identify young patients at high risk of POR.

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Published online 15 July 2014
Oocytes recovered after ovarian tissue slow freezing have impaired H2AX phosphorylation and functional competence 
Sam Sudhakaran, Shubhashree Uppangala, Sujith Raj Salian, Sachin D. Honguntikar, Ramya Nair, Guruprasad Kalthur and Satish Kumar Adiga

Ovarian tissue cryopreservation by slow freezing and vitrification are techniques available to cancer-affected women to preserve their fertility before cancer therapy. Comparison of these two techniques in mouse ovarian tissue has demonstrated the adverse effects of ovarian tissue slow freezing but not vitrification on oocyte DNA quality and functional ability. Hence, vitrification may be preferred over slow freezing for ovarian tissue cryopreservation.

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Published online 11 July 2014
Analysis and characterisation of bovine oocyte and embryo biomarkers by matrix-assisted desorption ionisation mass spectrometry imaging 
Roseli F. Gonçalves, Mónica S. Ferreira, Diogo N. de Oliveira, Rafael Canevarolo, Marcos A. Achilles, Daniela L. D'Ercole, Peter E. Bols, Jose A. Visintin, Gary J. Killian and Rodrigo R. Catharino

Lipid pathways play important biological roles in mammalian embryology. We document a new strategy to analyse bovine oocytes and embryos, and provide a chemical image using MALDI-MSI technology. Results obtained in this work demonstrates that a combination of MF, MSI features and chemometric analysis can be applied to discriminate cell stages, characterising specific biomarkers and relating them to developmental pathways.

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Published online 09 July 2014
Differential gene-expression profiles from canine cumulus cells of ovulated versus in vitro-matured oocytes 
Su-Jin Cho, Kyeong-Lim Lee, Yu-Gon Kim, Dong-Hoon Kim, Jae-Gyu Yoo, Byoung-Chul Yang, Jin-Ki Park and Il-Keun Kong

Canine oocytes are ovulated at the GV/GVBD stage and rely on CCs to mature to MII stage. Consequently, the role of CCs is very likely to be more important than in other species. The objective was to identify the relationship between oocytes and cumulus cells during oocyte maturation. The results demonstrated that cumulus cells play key roles in supporting oocyte maturation to the MII stage.

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Published online 08 July 2014
Semen collection, ejaculate characteristics and in vitro manipulation of spermatozoa from six species of captive flying-fox (Pteropus spp.) 
D. F. Melville, E. G. Crichton and S. D. Johnston

Almost two thirds of the 65 documented Pteropus species (large bats called flying-foxes) are considered threatened with extinction. Our research aimed to provide foundational information from which to develop a method of semen collection, extension and preservation for use in male fertility assessments and artificial insemination. The successful collection of >400 semen samples from six Pteropus species, four which had never been characterised before, adds to the general knowledge of male reproductive biology in these species and advances the likelihood that artificial reproductive technologies may one day be used to assist conservation efforts.

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Published online 26 June 2014
Impacts of and interactions between environmental stress and epigenetic programming during early embryo development 
Michael J. Bertoldo, Yann Locatelli, Christopher O'Neill and Pascal Mermillod

Embryos experience stress when they are grown in the laboratory, and this is believed to be detrimental to the embryo later in life. This article reviews the effects of in vitro conditions on the embryo and the mechanisms involved. By having a greater understanding of the mechanisms, we can potentially prevent stress during embryo growth in vitro.

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Published online 26 June 2014
Effects of varying doses of ?-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos) 
C. C. Stuart, J. L. Vaughan, C. M. Kershaw-Young, J. Wilkinson, R. Bathgate and S. P. de Graaf

Alpaca seminal plasma contains the protein β-nerve growth factor (β-NGF), which induces ovulation in females. We examined the effects of three doses of β-NGF and a gonadotrophin-releasing hormone (GnRH) agonist on ovulation rate and interval, corpus luteum diameter and plasma progesterone when administered to female alpacas and found that only ovulation rate varied. This suggests that there is no advantage to using β-NGF over more readily available and cheaper GnRH agonists to induce ovulation in alpacas.

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Published online 26 June 2014
Antibiotics and oxygen availability affect the short-term storage of spermatozoa from the critically endangered booroolong frog, Litoria booroolongensis 
Aimee J. Silla, Leesa M. Keogh and Phillip G. Byrne

Sperm storage technologies aim to extend sperm longevity and increase the time available to achieve artificial fertilisation. Overall, results from this study on the booroolong frog indicate that increased oxygen availability is beneficial to sperm longevity, but that antibiotics are detrimental. The optimal storage protocols identified will allow spermatozoa from spatially separated breeding stock to be transferred among captive facilities, a tool that will assist the genetic management of this critically endangered species.

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Published online 23 June 2014
Suboptimal maternal nutrition during early-to-mid gestation in the sheep enhances pericardial adiposity in the near-term fetus 
Shalini Ojha, Michael E. Symonds and Helen Budge

Changes in maternal diet during pregnancy can have long-lasting influences on the offspring’s health. In sheep, suboptimal nutrition during early-to-mid gestation results in enhanced fat deposition around the fetal heart. This is accompanied by increased abundance of genes that promote adipocyte synthesis. These adaptations may promote survival at birth but contribute to obesity in later life.

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Published online 19 June 2014
Effect of superstimulation protocols on nuclear maturation and distribution of lipid droplets in bovine oocytes 
D. Dadarwal, M. Honparkhe, F. C. F. Dias, T. Alce, C. Lessard and J. Singh

Lipids are the main energy source for oocytes. Based on 3D-microscopic evaluation, a mature bovine oocyte contained 22 picolitres of lipids distributed as 3000 droplets occupying 3.3% of the cell volume. Extending the FSH treatment from 4 to 7 days yielded a higher proportion of mature oocytes with no effect on lipids. FSH starvation for 84 h resulted in failure of meiotic resumption and accumulation of large lipid droplets, indicating degenerative changes.

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Published online 19 June 2014
Free-radical production after post-thaw incubation of ram spermatozoa is related to decreased in vivo fertility 
Enrique Del Olmo, Alfonso Bisbal, Olga García-Álvarez, Alejandro Maroto-Morales, Manuel Ramón, Pilar Jiménez-Rabadán, Luis Anel-López, Ana J. Soler, J. Julián Garde and María R. Fernández-Santos

This paper shows a direct relation between fertility and ROS production in sperm samples. The Fluorescein (CM-H2DCFDA) test combined with the SOF incubation could be considered a standard method for improving sperm-quality assessment. Ultimately, this technique could be a possible solution to the problem of fertility prediction in the laboratory.

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    | Supplementary Material (70 KB)
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Published online 16 June 2014
Insulin regulates primordial-follicle assembly in vitro by affecting germ-cell apoptosis and elevating oestrogen 
Xin-Lei Feng, Yuan-Chao Sun, Min Zhang, Shun-Feng Cheng, Yan-Ni Feng, Jing-Cai Liu, Hong-Hui Wang, Lan Li, Guo-Qing Qin and Wei Shen

Insulin plays an important role in the regulation of ovarian function; however, the specific molecular mechanism of its function remains largely unknown. This study found that insulin promoted folliculogenesis by facilitating germ-cell apoptosis within the cysts and upregulating oestrogen levels. This work established an effective model to study the function of insulin in folliculogenesis.

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    | Supplementary Material (285 KB)
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Published online 12 June 2014
Transgenerational inheritance of ovarian development deficiency induced by maternal diethylhexyl phthalate exposure 
Xi-Feng Zhang, Teng Zhang, Zhe Han, Jing-Cai Liu, Yu-Ping Liu, Jun-Yu Ma, Lan Li and Wei Shen

Diethylhexyl phthalate (DEHP) disrupts the physiological functions of endogenous hormones and induces abnormal development of mammals. However, the effects of DEHP exposure on folliculogenesis remain largely unknown. DEHP exposure significantly delayed the first meiotic progression of female fetal germ cells and accelerated follicle recruitment. DEHP exposure induced ovarian development deficiency, which was transgenerational in mice.

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    | Supplementary Material (581 KB)
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Published online 03 June 2014
Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay 
K. Pollock, J. Gosálvez, F. Arroyo, C. López-Fernández, M. Guille, A. Noble and S. D. Johnston

The integrity of sperm DNA is recognised as an important parameter of semen quality, but there are no published reports of this procedure for any amphibian. Validated as the first amphibian protocol for the assessment of sperm DNA fragmentation, the sperm chromatin dispersion test revealed three nuclear morphotypes representing varying degrees of DNA damage. This protocol has the potential to benefit assisted breeding programs using assisted reproductive technologies to support amphibian conservation efforts.

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Published online 29 May 2014
Open pulled straw vitrification and slow freezing of sheep IVF embryos using different cryoprotectants 
M. H. Bhat, V. Sharma, F. A. Khan, N. A. Naykoo, S. H. Yaqoob, G. Vajta, H. M. Khan, M. R. Fazili, N. A. Ganai and R. A. Shah

Embryo cryopreservation has huge potential to ensure the storage and worldwide distribution of elite germplasm. The aim of this study was to establish a standard procedure using specific cryoprotectant concentrations to avoid loss during cryopreservation procedures. Vitrification using 40% ethylene glycol resulted in superior embryo survival in sheep embryos from abattoir-derived oocytes. These results provide an easier option for embryo cryopreservation without compromising embryo survival.

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Published online 22 May 2014
Protein deubiquitination during oocyte maturation influences sperm function during fertilisation, antipolyspermy defense and embryo development 
Young-Joo Yi, Miriam Sutovsky, Won-Hee Song and Peter Sutovsky

Tagging with multi-ubiquitin chains targets substrate proteins for recycling by the 26S proteasome, during which deubiquitinating enzymes (DUBs) remove multi-ubiquitin chains from the substrate to regenerate mono-ubiquitin. In this study, DUB inhibitors impaired porcine oocyte maturation, fertilisation and pre-embryo development by altering the turnover of oocyte proteins such as major vault protein. Thus, DUBs may promote the acquisition of oocyte developmental competence.

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    | Supplementary Material (352 KB)
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Published online 28 April 2014
Distribution of cortical granules and meiotic maturation of canine oocytes in bi-phasic systems 
Maricy Apparicio, Giuliano Q. Mostachio, Tathiana F. Motheo, Aracelle E. Alves, Luciana Padilha, Eliandra A. Pires-Butler, Paula A. P. Savi, Ricardo A. R. Uscategui, Gaia C. Luvoni and Wilter R. R. Vicente

Studies about canine reproduction have increased over the years once this species can be used as a model for their endangered counterparts. This work aimed to reproduce in laboratory a similar hormonal condition in order to improve the results that are still far from satisfactory. Our results reveal that mimicking the hormonal changes is beneficial for oocyte maturation and so it is a more reliable approach to overcome the challenges of in vitro techniques.

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Published online 23 April 2014
Src family kinases are involved in the meiotic maturation of porcine oocytes 
Kateřina Kheilová, Jaroslav Petr, Tereza Žalmanová, Veronika Kučerová-Chrpová and Dalibor Řehák

Src family kinases are involved in the meiotic maturation of mammalian oocytes, but the information about their role in regulation of meiotic maturation has been mainly obtained from mouse oocytes. The present study proved that the Src family kinases play an active role in the meiotic maturation of porcine oocytes at least partly through MPF and MAPK activity regulation. Considering that data from mouse oocytes need not be validated in other species, our findings contribute to the understanding of the regulation of oocyte maturation.

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Published online 17 April 2014
Addition of l-ascorbic acid to culture and vitrification media of IVF porcine blastocysts improves survival and reduces HSPA1A levels of vitrified embryos 
Miriam Castillo-Martín, Marc Yeste, Albert Soler, Roser Morató and Sergi Bonet

The presence of non-enzymatic antioxidants in the environment surrounding the embryo reduces oxidative stress. Thus, the present study sought to determine how the addition of l-ascorbic acid during culture and vitrification and warming affects quality and gene expression of in vitro produced porcine blastocysts. Results obtained indicate that l-ascorbic acid supplementation increases embryo survival following cryopreservation via decreasing HSPA1A levels.

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Published online 10 April 2014
Substitution of starch for palm oil during gestation: impact on offspring survival and hepatic gene expression in the pig 
K. L. Almond, H. P. Fainberg, M. A. Lomax, P. Bikker, M. E. Symonds and A. Mostyn

Various dietary strategies to reduce piglet mortality have been tested, with increased fat in the maternal diet thought to improve energy partitioning to the fetus, subsequently improving neonatal outcome. However, we found that a maternal diet containing palm oil in place of starch led to reduced maternal glucose tolerance, increased neonatal mortality and altered gene expression in the liver at 1 week of age. Despite this, offspring from mothers fed the fat-substituted diet displayed no changes in postnatal growth or hepatic gene expression at 6 months of age.

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Published online 10 April 2014
In utero exposure to the oestrogen mimic diethylstilbestrol disrupts gonadal development in a viviparous reptile 
Laura M. Parsley, Erik Wapstra and Susan M. Jones

Studies of endocrine-disrupting chemicals (EDCs) in egg-laying reptiles have significantly advanced knowledge in this field; however, 30 % of reptiles are live-bearing. We examined the effects of feminising EDCs by exposing female Niveoscincus metallicus to the synthetic oestrogen, diethylstilbestrol, and found that gonadal development of male and female neonates from exposed mothers was impaired at birth. Maternal exposure to feminising EDCs may have profound consequences on reproductive fitness.

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Published online 10 April 2014
Antioxidant treatment during manipulation procedures prevents mitochondrial and DNA damage and enhances nuclear reprogramming of bovine somatic cell nuclear transfer embryos 
Hyo-Kyung Bae, In-Sun Hwang, Ji-Ye Kim, Sung-Young Lee, Choon-Keun Park, Boo-Keun Yang and Hee-Tae Cheong

The efficiency of animal cloning by somatic cell nuclear transfer (SCNT) is low because of incomplete nuclear reprogramming. We tried to prevent the mitochondrial and DNA damage caused by mechanical stress-associated reactive oxygen species during manipulation procedures of SCNT using antioxidants. Antioxidant treatment during manipulation procedures can efficiently prevent cellular damage and improve nuclear reprogramming.

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Published online 04 April 2014
Prenatal exposure to testosterone masculinises the female gerbil and promotes the development of lesions in the prostate (Skene 
Manoel F. Biancardi, Ana P. S. Perez, Cássia Regina Suzuki Caires, Rejane M. Góes, Patrícia S. L. Vilamaior, Fernanda C. A. Santos and Sebastião R. Taboga

This research reports important improvements in prostate biology focusing on the mechanisms underlying the origins of diseases from the female prostate (Skene’s gland). Thus, the aim of this study was to evaluate the interferential effects of exogenous testosterone exposure during the prenatal and pubertal periods on the health of the female prostate. We observed that the exposure to androgenic disruptors may predispose the gland to develop lesions with aging.

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Published online 03 April 2014
Differential expression of Notch component and effector genes during ovarian follicle and corpus luteum development during the oestrous cycle 
D. Murta, M. Batista, E. Silva, A. Trindade, L. Mateus, A. Duarte and L. Lopes-da-Costa

Ovarian dynamics involve extensive cell-to-cell communication, requiring fine regulation of cell signalling. This led us to evaluate the expression patterns of Notch cell-signalling pathway component and effector genes during follicle and corpus luteum development. We found Notch signalling active, and Notch expression spatially and temporally associated with key events in follicle and corpus luteum development. This suggests a relevant regulatory role of Notch signalling in ovarian dynamics.

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Published online 03 April 2014
Effects of vitrification on the expression of pluripotency, apoptotic and stress genes in in vitro-produced porcine blastocysts 
Miriam Castillo-Martín, Marc Yeste, Eva Pericuesta, Roser Morató, Alfonso Gutiérrez-Adán and Sergi Bonet

The demand for the successful preservation of porcine embryos has increased in the past decade. In this study, we evaluated the effects of vitrification and warming, a widely used cryopreservation method, on the expression of six relevant genes in in vitro-produced porcine blastocysts. The results suggest that vitrification and warming affect embryo developmental ability and pluripotency by increasing the number of cells with fragmented DNA, peroxide levels and HSPA1A gene expression.

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Published online 02 April 2014
Seasonal variation in urinary and salivary reproductive hormone levels in Amazonian manatees (Trichechus inunguis) 
Rodrigo S. Amaral, Fernando C. W. Rosas, Vera M. F. da Silva, Laura H. Graham, Priscila Viau, Marcilio Nichi and Claudio A. Oliveira

The Amazonian manatee is a threatened aquatic mammal endemic to the Amazon basin and the species is a seasonal breeder. The aim of this study was to evaluate the effect of the season on the reproductive hormone levels of captive Amazonian manatees. Different from males, all female hormones levels showed seasonal variation. The results strongly suggest the existence of reproductive seasonality in Amazonian manatees, even in captive conditions; however, apparently only females exhibit reproductive quiescence during the non-breeding season.

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Published online 31 March 2014
Effect of pregnancy on endometrial expression of luteolytic pathway components in the mare 
M. de Ruijter-Villani, H. T. A. van Tol and T. A. E. Stout

Preventing pulsatile endometrial PGF release is critical to pregnancy recognition in mares, and downregulation of PTGS2 and OXTR underlies initial avoidance of luteolysis. By Day 21 of pregnancy, however, expression of both PTGS2 and OXTR recovers to levels consistent with luteolysis. Continued luteal maintenance may instead involve uncoupling of PGF generating feedback loops by inhibition of PGF receptor (PTGFR) expression.

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Published online 31 March 2014
Ram spermatozoa migrating through artificial mucus in vitro have reduced mitochondrial membrane potential but retain their viability 
Carmen Martínez-Rodríguez, Mercedes Alvarez, Elena López-Urueña, Susana Gomes-Alves, Luis Anel-López, Cesar A. Chamorro, Luis Anel and Paulino de Paz

The ram sperm population that migrates through a column of cervical mucus surrogates in a penetration test was evaluated. These spermatozoa exhibit an adequate motility to cross the mucus barrier and we think that these cells can give information about the transport through the female genital tract. The viability of migrating spermatozoa was not changed but the mitochondrial status was affected after migration.

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Published online 27 March 2014
Cloning and expression of progesterone receptor isoforms A and B in bovine corpus luteum 
Robert Rekawiecki, Magdalena Karolina Kowalik and Jan Kotwica

Nuclear progesterone receptor has two main isoforms, PGRA and PGRB, which have different functions. PGRB cDNA from cattle was cloned then mRNA and protein levels for both isoforms in corpora lutea during the oestrous cycle and first trimester of pregnancy were determined. The data suggest that the variable expression of these isoforms during the oestrous cycle may depend on the influence of progesterone.

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Published online 25 March 2014
Contribution of boars to reproductive performance and paternity after homospermic and heterospermic artificial insemination 
C. E. R. Ferreira, D. B. Sávio, A. C. Guarise, M. J. Flach, G. D. A. Gastal, A. O. Gonçalves, O. A. Dellagostin, R. V. Alonso, I. Bianchi, C. D. Corcini and T. Lucia

Although it is commonly used in swine farms, heterospermic AI does not allow precise evaluation of boar fertility. The reproductive performance and the paternity contribution of four boars (A, B, C and D) were compared in this study using homospermic and heterospermic AI in 511 sows. Differences among boars were only identified after genotyping the piglets sired through heterospermic AI.

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Published online 25 March 2014
Validation of computer-assisted sperm-motility analysis in the amphibian Silurana tropicalis 
Severine Larroze, Daniel B. Pickford and William V. Holt

Computer-assisted sperm-motility analysis has been scarcely studied or used on amphibians. In this research we validated a standardised method producing reproducible results and compared traditional data analysis with sperm-subpopulation frequencies. We demonstrated that sperm-subpopulation frequency analysis can detect treatment effects on sperm motility.

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Published online 20 March 2014
Mouse ovarian tissue vitrification on copper electron microscope grids versus slow freezing: a comparative ultrastructural study 
Ferda Topal-Celikkan, Sinan Ozkavukcu, Deniz Balci, Sibel Serin-Kilicoglu and Esra Atabenli-Erdemli

Ovarian cryopreservation for fertility preservation is a gold standard procedure in numerous cases, like childhood cancers, although it is still considered experimental. Vitrification is widely used in embryo and oocyte cryopreservation, but its effectiveness for ovarian freezing remains debatable. Slow freezing seems to preserve ovarian content better and therefore the vitrification of tissues needs to be improved by means of cryoprotective agents and carrying devices.

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Published online 17 March 2014
Follistatin is essential for normal postnatal development and function of mouse oviduct and uterus 
S. J. Holdsworth-Carson, R. G. Craythorn, W. R. Winnall, K. Dhaliwal, R. Genovese, C. J. Nowell, P. A. W. Rogers, D. M. de Kretser, M. P. Hedger and J. E. Girling

Follistatin is a protein with key roles in the female reproductive system. We used mutant mouse strains to demonstrate the importance of follistatin in the postnatal development and function of the uterus and oviduct. Our work highlights the need to further examine the role of follistatin in the reproductive tract, as well as its key role in the ovary.

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    | Supplementary Material (1.3 MB)
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Published online 11 March 2014
Developmental programming of bone deficits in growth-restricted offspring 
Tania Romano, John D. Wark and Mary E. Wlodek

Offspring born with low birth weight are predisposed to a number of adult diseases, including osteoporosis. Herein, we report that such rat offspring develop reduced bone mineral content and strength at various ages. Importantly these deficits affected males more severely than females and became worse with aging. Interventions to improve bone quality in adult life for individuals born small may reduce the incidence of osteoporosis in the future.

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Published online 04 March 2014
Subpopulation pattern of eel spermatozoa is affected by post-activation time, hormonal treatment and the thermal regimen 
V. Gallego, M. C. Vílchez, D. S. Peñaranda, L. Pérez, M. P. Herráez, J. F. Asturiano and F. Martínez-Pastor

Eel culture requires closing of the life cycle in captivity. We have characterised sperm subpopulations in eel sperm by cluster analysis of CASA data. Keeping male eels at 20°C and using recombinant human chorionic gonadotropin (hCGrec) resulted in early and stable spermiation, with a higher proportion of a high quality subpopulation.

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    | Supplementary Material (512 KB)
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Published online 04 March 2014
Variation of spermatogenic and Sertoli cell number detected by fine needle aspiration cytology (FNAC) in Iberian red deer during and out of the breeding season 
Eliana Pintus, José Luis Ros-Santaella and José Julián Garde

Few studies have dealt with spermatogenesis in red deer, which represent a good model in which to investigate testicular function owing to their reproductive seasonality. Herein, we found that both spermatogenesis and Sertoli cell number decreased significantly out of the breeding season in Iberian red deer. Our results provide new insights into the dynamics of spermatogenesis and Sertoli cell populations, and challenge current dogma that Sertoli cell number is fixed in adulthood.

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Published online 28 February 2014
Sperm survival kinetics in different types of bull semen: progressive motility, plasma membrane integrity, acrosomal status and reactive oxygen species generation 
Mushtaq Ahmad, Nasim Ahmad, Amjad Riaz and Muhammad Anzar

Sperm survival is important for the fertilisation of oocytes. The kinetics of progressive sperm motility, acrosomes, plasma membrane and generation of reactive oxygen species were determined in vitro over 24 h. Bull sperm longevity was poor in frozen–thawed semen. Different types of spermatozoa followed different death pathways. All types of spermatozoa generated superoxide anions during incubation, which adversely affected their plasma membranes.

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Published online 24 February 2014
Selection of pre- versus postpubertal pig oocytes for parthenogenetic activation and somatic cell nuclear transfer 
H. S. Pedersen, Y. Liu, R. Li, S. Purup, P. Løvendahl, P. Holm, P. Hyttel and H. Callesen

Oocytes from sexually immature pigs are used for in vitro production techniques studying, for example, human diseases. The objective of the present study was met by developing a method for the selection of prepubertal oocytes with high developmental competence by measuring the oocyte’s diameter in combination with morphological evaluation. The study is important for understanding oocyte growth and development and for increasing the efficiency of in vitro work.

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Published online 24 February 2014
In vitro manipulation techniques of porcine embryos: a meta-analysis related to transfers, pregnancies and piglets 
Ying Liu, Juan Li, Peter Løvendahl, Mette Schmidt, Knud Larsen and Henrik Callesen

Over the last decade in vitro manipulation techniques of pig embryos have been used increasingly, but the overall tendencies and main problems are not well known. In this first systematic review we show that using more in vitro manipulations of porcine embryos decreases the overall production of piglets, and that nuclear transfer has developed markedly and now gives more stable results. This new knowledge illustrates the present status of in vitro techniques, and it will also be useful in the continuing attempts to identify problems and find solutions for nuclear transfer.

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Published online 21 February 2014
Effect of different freezing velocities on the quality and fertilising ability of cryopreserved rabbit spermatozoa 
E. Mocé, E. Blanch, A. Talaván and M. P. Viudes de Castro

The freezing step of the protocol for rabbit sperm cryopreservation negatively influences the performance of the spermatozoa. In this study we show the important role that the freezing rate plays in the quality and fertilising ability of the spermatozoa, with medium freezing rates (–40°C min–1) more suitable for freezing rabbit spermatozoa. These results could help to develop reliable protocols for rabbit sperm cryopreservation.

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Published online 19 February 2014
Elective single versus double embryo transfer: live birth outcome and patient acceptance in a prospective randomised trial 
Nicolás Prados, Rocío Quiroga, Cinzia Caligara, Myriam Ruiz, Víctor Blasco, Antonio Pellicer and Manuel Fernández-Sánchez

The aim of the study was to determine which strategy (i.e. single (eSET) versus double (eDET) embryo transfer) has a better trade-off in live birth delivery considering patient acceptance. The live birth delivery rate was significantly higher after eDET versus eSET (65% vs 42%, respectively), but no multiple births were observed after eSET compared with 35% after eDET. Nearly half the patients refused eSET even after having been well informed of its benefits.

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    | Supplementary Material (44 KB)
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Published online 19 February 2014
Oocyte expression, secretion and somatic cell interaction of mouse bone morphogenetic protein 15 during the peri-ovulatory period 
Brigitta Mester, Lesley J. Ritter, Janet L. Pitman, Adrian H. Bibby, Robert B. Gilchrist, Kenneth P. McNatty, Jennifer L. Juengel and C. Joy McIntosh

New work investigating how mammalian eggs achieve ovulation has found that the key fertility protein bone morphogenetic protein 15 (BMP15) is produced by the egg in more than one protein form just before ovulation. Whereas most previous studies focused on the mature region of BMP15, the present study shows that the proregion form is also increasingly produced just before ovulation and binds to surrounding cumulus and granulosa cells. This finding furthers our understanding of the complexity of BMP15 signalling and provides promising new routes of improving fertility treatments.

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Published online 14 February 2014
Knockdown of regulator of G-protein signalling 2 (Rgs2) leads to abnormal early mouse embryo development in vitro 
Yan Zhu, Ya-Hong Jiang, Ya-Ping He, Xuan Zhang, Zhao-Gui Sun, Man-Xi Jiang and Jian Wang

We hypothesised that Rgs2 may play a role in early embryo development. Therefore, we investigated the localisation of Rgs2 in mouse ovarian tissues and early embryos, and determined the effect of downregulating Rgs2 on the development of early embryos. Rgs2 was abundant in mouse early embryos, and the results of Rgs2 knockdown indicate that Rgs2 plays a critical role in early embryo development.

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Published online 12 February 2014
Polymorphisms in TOX and NCOA2 genes and their associations with reproductive traits in cattle 
Gregório M. F. de Camargo, Raphael B. Costa, Lucia G. de Albuquerque, Luciana C. A. Regitano, Fernando Baldi and Humberto Tonhati

The detection of causal mutation in genes improves genetic evaluation of animals. The aim of the present study was to search for causal mutations in the TOX and NCOA2 genes and relate them to reproductive traits in beef cattle. Two single nucleotide polymorphisms in the NCOA2 gene were associated (P < 0.05) with early pregnancy probability, age at first calving and days to first calving.

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Published online 11 February 2014
New insight into the castrated mouse epididymis based on comparative proteomics 
Xin Liu, Wenjuan Wang and FuJun Liu

The mammalian epididymis is responsible for sperm maturation and is critically regulated by testicular factors. Comparative proteomics was used to discover key testis-dependent proteins in epididymides from castrated mice. The results showed that the differentially expressed proteins participated primarily in energy metabolism and the antigen processing and presentation pathway. Our findings may provide potential new candidates for further studies into epididymal function and male aging.

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    | Supplementary Material (351 KB)
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Published online 30 January 2014
FSH and LH receptors are differentially expressed in cumulus cells surrounding developmentally competent and incompetent mouse fully grown antral oocytes 
Giulia Vigone, Valeria Merico, Carlo Alberto Redi, Giuliano Mazzini, Silvia Garagna and Maurizio Zuccotti

As experienced by many couples that undergo assisted reproduction, the oocyte quality is crucial for correct development. This study shows that Fsh and Lh receptors on the cumulus cells that surround the mouse oocyte are differentially expressed in developmentally competent or incompetent gametes. The method presented is a potential non-invasive procedure to discriminate between oocytes of good or bad quality.

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    | Supplementary Material (180 KB)
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Published online 30 January 2014
Shotgun proteomics of rainbow trout ovarian fluid 
Joanna Nynca, Georg J. Arnold, Thomas Fröhlich and Andrzej Ciereszko

Studies on the protein components of ovarian fluid (OF) may contribute to a better understanding of the mechanisms underlying oocyte development and quality. A shotgun proteomic approach was used to identify 54 proteins of in the OF of rainbow trout; most of the proteins are described for the first time in fish OF. The results of this study could help in the decoding of the biological function of these proteins and in the discovery of potential biomarkers of oocyte quality.

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    | Supplementary Material (18 KB)
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Published online 24 January 2014
Mitochondria and vesicles differ between adult and prepubertal sheep oocytes during IVM 
Karen L. Reader, Neil R. Cox, Jo-Ann L. Stanton and Jennifer L. Juengel

By identifying differences between good and poor quality oocytes (i.e. eggs), methods for improving oocyte quality may be ascertained. This research discovered differences in the distribution, number and structure of mitochondria and the volume of storage vesicles, between adult (good quality) and juvenile (poor quality) sheep oocytes. Treatments that can alter these cell organelles in the oocytes, either in the laboratory or the animal, may now be identified to improve oocyte quality.

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Published online 15 January 2014
Efficiency of delayed reproduction in Mus spicilegus 
Marie Lafaille, Patrick Gouat and Christophe Féron

To cope with varying ecological costs some species delay their first breeding, but delaying the first reproduction generally involves reproductive costs. We show here in a wild-type rodent (Mus spicilegus) that the delayed reproducers reproduce as well and more quickly than younger first-time breeders. The hypothesis of a lower reproductive potential in these relatively aged individuals seems to be contradicted.

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Published online 13 January 2014
Exogenous γ-tocotrienol promotes preimplantation development and improves the quality of porcine embryos 
Enok Lee, Sung-Hun Min, Bong-Seok Song, Ji-Yeong Yeon, Jin-Woo Kim, Jung-Ho Bae, Soo-Yong Park, Yong-Hee Lee, Sun-Uk Kim, Dong-Seok Lee, Kyu-Tae Chang and Deog-Bon Koo

Researchers have paid much attention to gamma-tocotrienol (GTT) due to its strong anti-oxidant properties. Therefore, the effects of GTT on blastocyst development, expression levels of reactive oxygen species and apoptotic index were investigated in preimplantation porcine embryos. We revealed that GTT improves the developmental competence and quality of porcine embryos by preventing oxidative stress.

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Published online 13 January 2014
Relationship between adiponectin and fertility in the female pig 
Danila B. Campos, Marcelo Albornoz, Paula C. Papa, Marie-France Palin, Vilceu Bordignon and Bruce D. Murphy

Adiponectin isoforms mediate different aspects of the pleiotropic function of the protein, including the reproductive process. We demonstrated that lower abundance of circulating adiponectin isoforms and altered ovarian adiponectin system expression are related to lower porcine fertility. These data present new elements contributing to a better understanding of the role of adiponectin in the reproduction and extends previous observations that reduced circulating adiponectin is associated with reproductive disorders, including PCOS. These findings open a new avenue for the application of commercial letrozole in monosex fish production.

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Published online 13 January 2014
Effect of dietary administration of letrozole and tamoxifen on gonadal development, sex differentiation and biochemical changes in common carp (Cyprinus carpio L.) 
Atul K. Singh, P. P. Srivastava, Rita Verma, Sharad C. Srivastava, Dinesh Kumar and Abubakar Ansari

Induced sex manipulation in fish is connected with better aquaculture production and sex control. Masculinisation in sexually undifferentiated common carp was attempted using non-steroidal tamoxifen and letrozole, which resulted in highly significant changes, particularly for letrozole, which produced almost all male C. capio. These findings open a new avenue for the application of commercial letrozole in monosex fish production.

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Published online 06 January 2014
Oxygen-regulated gene expression in murine cumulus cells 
Karen L. Kind, Kimberley K. Y. Tam, Kelly M. Banwell, Ashley D. Gauld, Darryl L. Russell, Anne M. Macpherson, Hannah M. Brown, Laura A. Frank, Daniel J. Peet and Jeremy G. Thompson

Oxygen is a key component of the environment during oocyte maturation, in vivo and in vitro. Expression of target genes of the hypoxia-inducible factors was increased in cumulus cells from mouse cumulus–oocyte complexes matured in vitro at 2% or 5%, compared with 20% oxygen. Oxygen-regulated gene expression is identified as a potential mechanism through which oxygen could influence cumulus cell function.

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Published online 03 January 2014
Aging alters histone H3 lysine 4 methylation in mouse germinal vesicle stage oocytes 
Gen-Bao Shao, Jie Wang, Liu-Ping Zhang, Chao-Yang Wu, Jie Jin, Jian-Rong Sang, Hong-Yan Lu, Ai-Hua Gong, Feng-Yi Du and Wan-Xin Peng

Abnormalities of chromatin modifications in old MII oocytes are the leading cause of mammalian infertility. Recent studies report that changes in histone acetylation at the MII stage arise at the GV stage. In the present study, aging caused abnormalities of histone methylation and demethylase at the GV stage. Inhibition of the demethylase alleviated some of the aging-caused GV abnormalities. These results provide insights for future studies into the effects of reproductive aging on chromatin modifications.

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Published online 03 January 2014
T26248G-transversion mutation in exon7 of the putative methyltransferase Nsun7 gene causes a change in protein folding associated with reduced sperm motility in asthenospermic men 
Nahid Khosronezhad, Abasalt Hosseinzadeh Colagar and Syed Golam Ali Jorsarayi

The Nsun7 gene with location 4p14, encodes putative methyltransferase Nsun7, has a role in sperm motility. Analysis of the direct sequencing of PCR products of exon7 showed a T26248G-transversion mutation with protein folding and function exchange in asthenospermic men, which may lead to reduced sperm motility and infertility. Based on, it can be helpful as an infertility marker for males with a history of subfertility or idiopathic infertility in fertility assessment.

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Published online 03 January 2014
Assessment of DNA damage in goat preantral follicles after vitrification of the ovarian cortex 
Luciana R. Faustino, Adeline A. Carvalho, Cleidson M. G. Silva, Rafael Rossetto, Cláudio A. P. Lopes, Maurício F. van Tilburg, Pedro B. M. Carneiro, Sônia N. Báo, Arlindo A. A. Moura, Vilceu Bordignon, José R. Figueiredo and Ana Paula R. Rodrigues

Effective methods for tissue cryopreservation should protect cell viability and genome integrity. This study used the TUNEL assay and detection of γH2AX foci to investigate DNA damage in goat ovarian tissues cryopreserved with a new vitrification method (the Ovarian Tissue Cryosystem). This vitrification method preserved both morphology and DNA integrity of goat preantral follicles and therefore represents a potential alternative for ovarian tissue preservation in other species.

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Published online 23 December 2013
Interaction between differential gene expression profile and phenotype in bovine blastocysts originating from oocytes exposed to elevated non-esterified fatty acid concentrations 
V. Van Hoeck, D. Rizos, A. Gutierrez-Adan, I. Pintelon, E. Jorssen, I. Dufort, M. A. Sirard, A. Verlaet, N. Hermans, P. E. J. Bols and J. L. M. R. Leroy

Metabolic disorders are risk factors for reproductive failure and elevated non-esterified fatty acid (NEFA) concentrations have been proposed as key contributors by affecting oocyte quality. The aim of this study was to unravel the underlying molecular mechanisms through which NEFAs impact on bovine maturing oocytes and subsequent embryo development. Transcriptome and functional data point to metabolic dysregulation in the resulting preimplantation embryos, findings that might explain the reduced conception rates in females across species suffering lipolytic disorders.

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Published online 20 December 2013
Supplementation with omega-3 fatty acids during gestation and lactation to a vitamin B12-deficient or -supplemented diet improves pregnancy outcome and metabolic variables in Wistar rats 
Amrita Khaire, Richa Rathod, Nisha Kemse, Anvita Kale and Sadhana Joshi

Maternal vitamin B12 deficiency is known to lead to an adverse pregnancy outcome and increase the risk for developing diseases in mothers in later life. This study examines the effect of maternal n-3 PUFA supplementation to vitamin B12-deficient or -supplemented diets on pregnancy outcome, fatty-acid status and metabolic variables in Wistar rats. Results indicate that maternal vitamin B12 status is critical in determining pregnancy outcome and metabolic variables in dams and supplementation with n-3 PUFA is beneficial.

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Published online 17 December 2013
Dynamic changes in nuclear import of a nuclear localisation signal-bearing substrate in 8-cell stage porcine embryos 
Yanfang Li, Ki-Eun Park and Ryan A. Cabot

Mechanisms that partition intracellular proteins between the nucleus and cytoplasm have been shown to be critical players in cellular differentiation. Using the porcine embryo as our model, we found that the activity of discrete nuclear import pathways changes during progression from the oocyte to the 8-cell stage embryo. This suggests that nuclear import pathways serve a critical role regulating cleavage development.

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Published online 12 December 2013
Influence of pathogenic bacteria species present in the postpartum bovine uterus on proteome profiles 
A. M. Ledgard, G. A. Smolenski, H. Henderson and R. S.-F. Lee

After parturition >90% of dairy cows will have some form of uterine infection with the risk of developing endometritis, which can reduce conception rates. We examined the effect of uterine infection or inflammation on the postpartum uterus and identified 12 proteins that correlated with the presence of bacterial pathogens and polymorphonuclear neutrophils. These indicator proteins could be used as biomarkers of subclinical endometritis.

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Published online 04 December 2013
Progestin priming before gonadotrophin stimulation and AI improves embryo development and normalises luteal function in the cat 
Rosemary A. Stewart, Adrienne E. Crosier, Katharine M. Pelican, Budhan S. Pukazhenthi, Brandon D. Sitzmann, Tom E. Porter, David E. Wildt, Mary Ann Ottinger and JoGayle Howard

Exogenous gonadotrophins can alter endocrine dynamics and inhibit embryo development in felids. We demonstrate that priming the cat ovary with oral progestin before gonadotrophin stimulation and AI mitigates the negative influence of gonadotrophin therapy by normalising luteal function and improving embryonic developmental competence. These data enhance our understanding of the mechanisms involved in cat ovarian control and have practical implications for refining assisted reproductive technologies in endangered felids.

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Published online 04 December 2013
Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability 
I. Ortiz, J. Dorado, D. Acha, M. J. Gálvez, M. Urbano and M. Hidalgo

The Andalusian donkey is an endangered breed and sperm banks are needed; however, cryopreservation may result in low sperm quality. This study investigated the effect of colloid single-layer centrifugation (SLC) on frozen–thawed donkey semen. It concluded that SLC is a suitable procedure to improve cryopreserved donkey semen, in particular for low-freezability ejaculates. These results could be helpful in the conservation of endangered donkey breeds.

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Published online 27 November 2013
The beneficial effects of reduced magnesium during the oocyte-to-embryo transition are conserved in mice, domestic cats and humans 
Jason R. Herrick, Kevin J. Strauss, Ann Schneiderman, Mary Rawlins, John Stevens, William B. Schoolcraft and Rebecca L. Krisher

Reduced concentrations of Mg2+ during in vitro fertilisation or intracytoplasmic sperm injection improved cleavage or embryonic development in mice, domestic cats and humans. Although specific mechanisms may vary between species, sensitivity to increased concentrations of Mg2+ in the medium during the oocyte-to-embryo transition appears to be conserved in three different mammalian species.

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Published online 27 November 2013
Influence of sperm fertilising concentration, sperm selection method and sperm capacitation procedure on the incidence of numerical chromosomal abnormalities in IVF early bovine embryos 
Sebastián Demyda-Peyrás, Jesús Dorado, Manuel Hidalgo and Miguel Moreno-Millán

Chromosomal abnormalities are primary contributors to reduced efficiency of in IVF embryos, having a negative effect on the success of the entire process. Herein, we demonstrate that sperm fertilising concentration and handling increase the appearance of such chromosomal abnormalities. By using a lower sperm concentration and an adequate capacitation compound, the effect of chromosomal abnormalities in IVF embryos can be reduced.

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Published online 12 November 2013
Structure, histochemistry and seasonal variations of the male reproductive accessory glands in the Pallas 
Caroline M. Christante, Mateus R. Beguelini, Cintia C. I. Puga, Ana C. Negrin, Eliana Morielle-Versute, Patrícia S. L. Vilamaior and Sebastião R. Taboga

The male reproductive accessory glands (RAGs) of bats vary widely according to the reproductive strategies of each species. Thus, this study aimed to characterise the RAGs of Molossus molossus (Molossidae) and evaluate their seasonal variations. Results demonstrated the existence of two regions (ventral and dorsal), which had inherent characteristics; however, both synchronised to establish the main reproductive peak in spring (early rainy season).

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Published online 11 November 2013
Tolerance of spermatozoa to hypotonic stress: role of membrane fluidity and correlation with cryosurvival 
Harriëtte Oldenhof, Anna Heutelbeck, Anne-Kathrin Blässe, Heinrich Bollwein, Gunilla Martinsson, Willem F. Wolkers and Harald Sieme

Sperm from different stallions exhibit great differences in cryosurvival rates, which likely can be attributed to differences in cellular properties, such as membrane stability towards osmotic stress. The aim of this study was to obtain insights in factors determining membrane stability as well as cryosurvival. Pre-freeze osmotic tolerance was evaluated for individual stallions, and it was found that this cannot be used as a parameter to predict sperm cryosurvival rates.

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Published online 11 November 2013
Exposure to Brefeldin A promotes initiation of meiosis in murine female germ cells 
Lian-Jun Zhang, Bo Chen, Xin-Lei Feng, Hua-Gang Ma, Li-lan Sun, Yan-Min Feng, Gui-Jin Liang, Shun-Feng Cheng, Lan Li and Wei Shen

Our research focused on the mechanism of meiosis initiation, a complex process that is still not clear. In this study, we cultured 11.5 days post coitum female mouse genital ridges in vitro with exposure to Brefeldin A and found that meiosis initiation was stimulated partly via a retinoic acid–Stra8 signal. Our results contributed to the study of meiosis initiation in vitro and also confirmed the role of Stra8 in meiosis initiation.

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    | Supplementary Material (336 KB)
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Published online 06 November 2013
Follicle size and reproductive hormone profiles during a post-weaning altrenogest treatment in primiparous sows 
J. J. J. van Leeuwen, M. R. T. M. Martens, J. Jourquin, M. A. Driancourt, A. Wagner, B. Kemp and N. M. Soede

Post-weaning treatments with a progesterone analogue can be applied in sows to postpone oestrus after weaning, which may affect subsequent fertility. The present study investigated the underlying endocrinology of this phenomenon. Daily altrenogest treatment resulted in a wave of follicle growth for 5 days and a concomitant increase in FSH, but follicles lost oestrogenic competence by Day 2. The latter may be the cause of reduced fertility after short altrenogest treatments.

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Published online 01 November 2013
Effect of vehicle and route of administration of letrozole on ovarian function in a bovine model 
M. J. Yapura, R. J. Mapletoft, R. A. Pierson, J. Singh and G. P. Adams

The use of natural or synthetic oestrogens for controlling reproductive function has been the subject of considerable controversy in the cattle industry. Letrozole, an aromatase inhibitor, prevents the body from producing its own oestrogens. The ability of aromatase inhibitors to control follicular dynamics holds potential for development of letrozole-based (non-steroidal) methods of controlling and synchronising ovarian function in cattle and other species.

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Published online 30 October 2013
Structure, histochemistry, ultrastructure and seasonal variations of the male prostatic complex in the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae) 
Ana C. Negrin, Mateus R. Beguelini, Cintia C. I. Puga, Caroline M. Christante, Larissa M. Bueno, Eliana Morielle-Versute, Patrícia S. L. Vilamaior and Sebastião R. Taboga

Because of its wide geographic distribution, Myotis nigricans exhibit considerable variability in reproductive strategies. Thus, the aim of the present study was to characterise the prostatic complex of M. nigricans and evaluate its seasonal variations. The prostatic complex is composed of three bilobed regions (ventral, dorsolateral and dorsal), each with distinctive morphological characteristics. However, all three regions synchronise to establish the main reproductive peak in summer.

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Published online 25 October 2013
Recovery of fertility from adult ovarian tissue transplanted into week-old Japanese quail chicks 
Jianan Liu, Kimberly M. Cheng and Frederick G. Silversides

Fertility can be recovered from birds’ ovaries using transplantation. Fragments of ovaries from adult Japanese quail were transplanted into week-old quail recipients and chicks were produced from the donor tissue, demonstrating that adult birds can be used as ovarian donors for genetic conservation. Changes in egg production with the transplants also suggested that ovarian transplantation can be used to study reproduction in birds.

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Published online 24 October 2013
EZH2 is essential for development of mouse preimplantation embryos 
Xian-Ju Huang, Xuguang Wang, Xueshan Ma, Shao-Chen Sun, Xiaolong Zhou, Chengcheng Zhu and Honglin Liu

Ezh2 controls the fate of mouse embryos at an early stage. We investigated Ezh2 expression during preimplantation and found that lacking Ezh2 results in mouse embryos with severe development defects. Our results showed that Ezh2 affect mouse embryo development through the regulation of epigenetic modification, gene expression and apoptosis.

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   | Supplementary Material (622 KB)
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Published online 24 October 2013
The pattern of LH secretion and the ovarian response to the ‘ram effect’ in the anoestrous ewe is influenced by body condition but not by short-term nutritional supplementation 
R. J. Scaramuzzi, L. Oujagir, J-B. Menassol, S. Freret, A. Piezel, H. M. Brown, J. Cognié and C. Fabre Nys

The food animals eat has a major influence on their ability to reproduce. This experiment was designed to determine if fatness or high-energy diets affected the socio-sexual responses of ewes to rams. High-energy diets stimulated insulin and glucose but had little influence on reproductive responses. Thin ewes had reduced glucose and insulin and compromised responses to socio-sexual stimulation, suggesting that fatness was a significant contributor to reproductive success.

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Published online 21 October 2013
Y chromosome in Turner syndrome: detection of hidden mosaicism and the report of a rare X;Y translocation case 
Adriana Valéria Sales Bispo, Pollyanna Burégio-Frota, Luana Oliveira dos Santos, Gabriela Ferraz Leal, Andrea Rezende Duarte, Jacqueline Araújo, Vanessa Cavalcante da Silva, Maria Tereza Cartaxo Muniz, Thomas Liehr and Neide Santos

The presence of Y-chromosome material in Turner syndrome is of medical importance because it results in an increased risk of developing gonadal tumours and virilisation. We detected cryptic Y-chromosome sequences in 2.7% of cases and describe one girl with a very uncommon karyotype 46,X,der(X)t(X;Y)(p22.3?2;q11.23). We recommend that the search for Y-chromosome mosaicism should be carried out in all Turner cases.

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Published online 21 October 2013
Are the basal cells of the mammalian epididymis still an enigma? 
S. Arrighi

This review summarises the striking novelties obtained from morphophysiological investigations after David W. Hamilton stated ‘epididymal basal cells are an enigma’ in the 1970s. In addition to an earlier hypothesised scavenger role linked to the expression of macrophage antigens, basal cells have been shown to be involved in cell–cell cross-talk and to function as luminal sensors regulating epithelial activity.

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Published online 21 October 2013
Spermatozoa of the old endemic rodents of Australia – the possible functional significance of their ventral processes 
Simon Drew, Chris Leigh and William G. Breed

Most Australian old endemic rodents contain a highly complex spermatozoon with the head containing two processes extending from its upper concave surface. This study investigates the structure and function of these processes.  It shows that they are highly rigid structures and suggests that they may stabilise the spermatozoon at the time of egg-coat binding. Since these spermatozoa occur throughout this rodent group they probably evolved in an early common ancestor over 5 million years ago.

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Published online 25 September 2013
Isolation, characterisation and cDNA sequencing of a new form of parvalbumin from carp semen 
Mariola A. Dietrich, Błażej Westfalewicz, Patrycja Jurecka, Ilgiz Irnazarow and Andrzej Ciereszko

Parvalbumins are calcium-binding proteins and their presence in carp semen may be related to the importance of calcium ions in the mechanism controlling carp sperm movement. We aimed to purify and characterise the new form of carp seminal parvalbumin (Pv-II), which differed from other carp parvalbumins and had a tendency to oligomerise. The oligomerisation was protected by the presence of seminal plasma. The results suggest the role of parvalbumin in mechanism of carp sperm activation and spermatozoa protection against apoptosis.

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Published online 18 September 2013
LAPS-FSH: a new and effective long-acting follicle-stimulating hormone analogue for the treatment of infertility 
Sunyoung Jung, Youngjin Park, YoungHoon Kim, Yu Yon Kim, Hyun-Ji Choi, Woo-Chan Son and SeChang Kwon

As FSH has relatively short half-life, there is a great need for the development of a long-acting FSH analogue with increased efficacy. A new FSH analogue, LAPS-FSH, demonstrates several advantages such as a long half-life, good potency, and ability to increase the number of Graafian follicles and the size of the follicles. LAPS-FSH is expected to increase the success rate of infertility treatments while also reducing the effort and time.

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Published online 17 September 2013
Altered chromatin condensation of heat-stressed spermatozoa perturbs the dynamics of DNA methylation reprogramming in the paternal genome after in vitro fertilisation in cattle 
Mohammad Bozlur Rahman, Md. Mostofa Kamal, Tom Rijsselaere, Leen Vandaele, Mohammed Shamsuddin and Ann Van Soom

Shortly after fertilisation, methylation of both maternal and paternal DNA is reprogrammed for further embryo development. The objectives of this study were to investigate DNA methylation reprogramming following fertilisation of oocytes with heat-stressed spermatozoa and the results show that the normal pattern of DNA methylation reprogramming is perturbed in the paternal genome, leading to a lower fertilisation rate. Our data can be used to address lower fertilisation potential of spermatozoa with suboptimal quality.

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Published online 12 September 2013
Cryotolerance and global gene-expression patterns of Bos taurus indicus and Bos taurus taurus in vitro- and in vivo-produced blastocysts 
Mateus J. Sudano, Ester S. Caixeta, Daniela M. Paschoal, Alicio Martins, Rui Machado, José Buratini and Fernanda D. C. Landim-Alvarenga

The major challenge for a greater dissemination of in vitro-produced bovine embryos is to improve embryonic survival after cryopreservation. The results generated in this study provide different transcriptomic signatures of a model with two bovine subspecies and two embryo origins that presented distinct lipid content and post-cryopreservation survival. Such knowledge will be useful to characterise the specific role of identified genes in lipid metabolism and embryo cryosurvival capacity.

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Published online 06 September 2013
Predictive value of antral follicle count and anti-Müllerian hormone for follicle and oocyte developmental competence during the early prepubertal period in a sheep model 
Laura Torres-Rovira, Antonio Gonzalez-Bulnes, Sara Succu, Antonio Spezzigu, Maria E. Manca, Giovanni G. Leoni, Marina Sanna, Salvatore Pirino, Marilia Gallus, Salvatore Naitana and Fiammetta Berlinguer

Factors and mechanisms driving early prepubertal follicle development still need to be completely elucidated. Here, we found that antral follicle count (AFC) could represent a sign of follicular maturation in the prepubertal ovaries, while its close relationship with the non-growing follicle pool could develop only after the full establishment of reproductive activity. These data extend our knowledge of the ovarian functionality during the early prepubertal period.

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Published online 28 August 2013
Triphenyltin chloride induces spindle microtubule depolymerisation and inhibits meiotic maturation in mouse oocytes 
Yu-Ting Shen, Yue-Qiang Song, Xiao-Qin He, Fei Zhang, Xin Huang, Yu Liu, Lu Ding, Lin Xu, Mao-Bi Zhu, Wen-Feng Hu, Zhong-Quan Qi, Hai-Long Wang and Xiang-Jun Yang

Triphenyltin chloride (TPTCL) is a highly bioaccumulated and toxic environmental oestrogen; however, its effect on oocyte meiosis remains unknown. In this study we examined the effect of TPTCL on mouse oocyte meiotic maturation in vitro and in vivo and found that TPTCL seriously affects meiotic maturation by disturbing cell-cycle progression and the microtubule cytoskeleton. Our study may be helpful for investigating infertility and miscarriage of undetermined origin.

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Published online 27 August 2013
Mitochondrial SIRT5 is present in follicular cells and is altered by reduced ovarian reserve and advanced maternal age 
Leanne Pacella-Ince, Deirdre L. Zander-Fox and Michelle Lane

Young women whose eggs age prematurely or women over 40 have reduced fertility. Our study shows that the follicular cells of these women have reduced activity of a mitochondrial protein, SIRT5, that alters the activity of enzymes resulting in an accumulation of ammonium in the follicular fluid that bathes the egg. This altered follicular-fluid composition may contribute to the reduced egg quality in these women.

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blank image Reproduction, Fertility and Development
Volume 26 Number 7 2014

 
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Three-dimensional systems for in vitro follicular culture: overview of alginate-based matrices 
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Ivina R. Brito , Isadora M. T. Lima , Min Xu , Lonnie D. Shea , Teresa K. Woodruff and José R. Figueiredo
pp. 915-930

In vitro follicle culture can be accomplished using two- or three-dimensional systems. This review focuses on three-dimensional in vitro culture systems based on the use of alginate hydrogel and its importance for the maintenance of follicular architecture, which is essential to ensure appropriate interactions between granulosa cells and the oocyte. Such a culture system has many important applications including its use in assisted reproductive technology.

 
  
 

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Pentose phosphate pathway activity: effect on in vitro maturation and oxidative status of bovine oocytes 
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Cynthia Gutnisky , Gabriel C. Dalvit , Jeremy G. Thompson and Pablo D. Cetica
pp. 931-942

To improve embryo in vitro production techniques, is necessary to understand the variables involved in the process. In the present study, we investigated the control of the pentose phosphate pathway (PPP), a key metabolic pathway involved in NADPH and ribose synthesis, in cumulus–oocytes complexes during IVM and revealed a link between PPP activity and oxidative and mitochondrial metabolism of the oocyte. These findings will help us understand the relationship between the PPP in cumulus cells and events related with the maturational capability of the oocyte.

 
  
 

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In vitro development of preimplantation porcine embryos using alginate hydrogels as a three-dimensional extracellular matrix 
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Catherine N. Sargus-Patino , Elane C. Wright , Sarah A. Plautz , Jeremy R. Miles , Jeff L. Vallet and Angela K. Pannier
pp. 943-953

Deficiencies in porcine embryo elongation contribute to approximately 20% of embryonic loss, but exact mechanisms of elongation are poorly understood. We have established a culture system using alginate hydrogel as a three-dimensional matrix that facilitates morphological changes with increased oestrogen production and differential gene expression of porcine embryos, consistent with in vivo elongation. This alginate culture system can serve as a tool for evaluating specific mechanisms of embryo elongation.

 
  
 

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Equine oviduct explant culture: a basic model to decipher embryo–maternal communication 
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Hilde Nelis , Katharina D’Herde , Karen Goossens , Lynn Vandenberghe , Bart Leemans , Katrien Forier , Katrien Smits , Kevin Braeckmans , Luc Peelman and Ann Van Soom
pp. 954-966

The subtle manner of embryo–oviductal communications in the horse is very difficult to analyse in vivo. Therefore, we developed a system sustaining equine oviduct explants, bordered by highly differentiated, functional and intact epithelial cells, with only a negligible percentage of apoptotic or necrotic cells and so mimicking the in vivo situation very closely. Since the system is easy to set up, it appears to be an excellent tool for proteome, transcriptome and miRNome analysis, in order to unravel embryo–maternal interactions in the horse.

 
  
 

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Gestational protein restriction delays prostate morphogenesis in male rats 
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Cristiane F. Pinho , Mariana A. Ribeiro , Jaqueline C. Rinaldi , Sergio L. Felisbino , Patricia F. Pinheiro , Raquel F. Domeniconi , Ricardo A. Fochi , Patrícia A. Boer and Wellerson R. Scarano
pp. 967-973

Maternal nutritional status during pregnancy is essential for normal growth, as intra-uterine development is dependent on the nutritional, hormonal and metabolic environment provided by the mother. We investigated the effects of gestational protein restriction on prostate morphogenesis in male rats and the results showed that maternal malnutrition delayed prostate development. This result could be an important factor in altered postnatal development and long-term diseases in the prostate.

 
  
 

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Induction of autophagy during in vitro maturation improves the nuclear and cytoplasmic maturation of porcine oocytes 
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Bong-Seok Song , Ji-Su Kim , Young-Hyun Kim , Bo-Woong Sim , Seung-Bin Yoon , Jae-Jin Cha , Seon-A Choi , Hae-Jun Yang , Seong-Eun Mun , Young-Ho Park , Kang-Jin Jeong , Jae-Won Huh , Sang-Rae Lee , Sang-Hyun Kim , Sun-Uk Kim and Kyu-Tae Chang
pp. 974-981

The critical role of autophagy has been demonstrated in mammalian early embryogenesis. The present study was conducted to elucidate the effect of rapamycin, a chemical autophagy inducer, on the nuclear and cytoplasmic maturation of porcine oocytes. We revealed that autophagy induction by rapamycin could improve the rate of MII oocyte production, monospermic fertilization, blastocyst formation rate, total cell number and cell survival.

 
  
 

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Ovarian hyperstimulation affects fluid transporters in the uterus: a potential mechanism in uterine receptivity 
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Laura A. Lindsay and Christopher R. Murphy
pp. 982-990

Controlled ovarian hyperstimulation, commonly involved in fertility treatment, is associated with an altered uterine environment, making it less receptive to an implanting blastocyst. A rat ovarian hyperstimulation model used in this study showed an increase in AQP5 water channels and a decrease in a tight junction protein, claudin-4, in the uterus at the time of implantation. These results suggest that there is an alteration in uterine luminal fluid dynamics and content at the time of implantation in the ovarian hyperstimulated rat.

 
  
 

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Effects of castration on androgen receptors and gonadotropins in the pituitary of adult male viscachas 
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Verónica Filippa , Daiana Godoy , Edith Perez and Fabian Mohamed
pp. 991-1000

Androgens are essential for the initiation and maintenance of reproduction in mammals. Androgens act via androgen receptors (AR) and can regulate the function of a variety of cells in a number of organs, including pituitary cells secreting gonadotropins (LH and FSH). In the present study we saw a decrease in pituitary cells expressing AR, LH and FSH, and in cells coexpressing FSH and AR, in castrated male viscachas. These findings suggest that androgens could differentially affect the LH-and FSH-secreting cells.

 
  
 

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Human oocyte vitrification with corona radiata, in autologous follicular fluid supplemented with ethylene glycol, preserves conventional IVF potential: birth of four healthy babies 
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Xian-Hong Tong , Li-Min Wu , Ren-Tao Jin , Hong-Bing Luan and Yu-Sheng Liu
pp. 1001-1006

There is the possibility for a woman to cryopreserve and store her own eggs in an egg bank until she is ready to proceed with motherhood. Autologous follicular fluid supplemented with ethylene glycol and sucrose was an efficient cryoprotectant for egg cryopreservation; the frozen–thawed eggs have similar potential for fertilisation and development as that of fresh eggs. This simple, feasible method for egg cryopreservation can be popularised in all reproductive centres.

 
  
 

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Synthesis of leukotrienes in porcine uteri with endometritis induced by infection with Escherichia coli 
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Barbara Jana , Joanna Czarzasta and Jerzy Jaroszewski
pp. 1007-1016

Leukotrienes (LTs) are pro-inflammatory lipid mediators and their production in inflamed uteri is not fully understood. The present experiment aimed to determine LTB4 and LTC4 amounts and 5-lipooxygenase, LTA4 hydrolase and LTC4 synthase mRNA levels and protein expression in the porcine uteri with inflammation. The data obtained show the upregulation of LT synthesis and secretion in inflamed porcine uteri, which suggests the importance of these factors for functions of pathologically changed organs.

 
  
 

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Long-term effect on in vitro cloning efficiency after treatment of somatic cells with Xenopus egg extract in the pig 
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Ying Liu , Olga Østrup , Rong Li , Juan Li , Gábor Vajta , Peter M. Kragh , Mette Schmidt , Stig Purup , Poul Hyttel , Dan Klærke and Henrik Callesen
pp. 1017-1031

Incomplete reprograming of somatic cells is one of the important reasons for the low efficiency of somatic cell nuclear transfer (SCNT). In our study we investigated the reprogramming effect of Xenopus egg extract on somatic donor cells from pig used subsequently for SCNT, and our results showed that a single extract treatment produces a long-term increase in in vitro efficiency of SCNT. Our findings provide new understandings about the process of cellular reprogramming and also have potential application for future work on practical SCNT.

 
    | Supplementary Material (25 KB)
 

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Transgenerational programming of fetal nephron deficits and sex-specific adult hypertension in rats 
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Linda A. Gallo , Melanie Tran , Luise A. Cullen-McEwen , Kate M. Denton , Andrew J. Jefferies , Karen M. Moritz and Mary E. Wlodek
pp. 1032-1043

A suboptimal pregnancy environment that reduces birth weight has the potential to program disease in subsequent generations. The aim of the study was to examine transgenerational cardio-renal effects, via the maternal line, using a rat model of uteroplacental insufficiency. We report alterations to kidney development and transient, sex-specific hypertension in second generation offspring. This study demonstrates that in utero exposures can have far-reaching effects that extend into the next generation.

 
  
 

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Human seminal fibronectin fragmentation patterns and their domain immunoreactivities in leucocytospermic patients 
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Ewa M. Kratz , Marcin Wójtowicz , Magdalena Przybysz , Ricardo Faundez and Iwona Kątnik-Prastowska
pp. 1044-1051

Male infertility has become a social and medical problem of developed countries over the last decades. This work reports the differences in degree of fibronectin fragmentation and its domain reactivities in seminal plasmas of fertile normozoospermic and infertile leucocytospermic male patients. It is expected that the analysis of human seminal fibronectin molecular status would be helpful for selecting the highest quality spermatozoa for use in assisted reproduction techniques.

 
  
 

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FOXM1 is lower in human fetal membranes after spontaneous preterm labour and delivery 
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Ratana Lim , Gillian Barker and Martha Lappas
pp. 1052-1060

Preterm birth is typically associated with infection and/or inflammation, and being accountable for approximately 1 million annual neonatal deaths, is a global healthcare issue. We found that spontaneous preterm labour is associated with a decrease in transcription factor FOXM1 in fetal membranes, with FOXM1 gene silencing in primary amnion cells increasing inflammation-induced pro-labour mediators. The role of FOXM1 in labour processes enhances our understanding of the mechanisms that drive preterm birth, perhaps leading to a future clinical therapeutic.

 
  
 

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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.

    RD14089  Accepted 25 September 2014
    Stable reference genes in granulosa cells of bovine dominant follicles during follicular growth, FSH stimulation and maternal aging
    Muhammad Khan, Fernanda Dias, Isabelle Dufort, Vikram Misra, Sirard Marc-andré, Jaswant Singh
    Abstract


    RD14346  Accepted 25 September 2014
    Controlled hydrostatic pressure stress downregulates the expression of ribosomal genes in preimplantation embryos - a possible protection mechanism?
    Istvan Bock, Hadas Amit, Eszter Losonczi, Ana Carstea, Anita Feher, Kaveh Mashayekhi, Szabolcs Matyas, Andras Dinnyes, Csaba Pribenszky
    Abstract


    RD14186  Accepted 24 September 2014
    Association of C677T Transition of the Human Methylenetetrahydrofolate Reductase Gene with Male Infertility
    Mohammad Karimian, Abasalt Hosseinzadeh Colagar
    Abstract


    RD14294  Accepted 23 September 2014
    Localisation and endocrine control of HAS2, HAS3 and CD44 expression in sheep granulosa cells
    Ramyar Chavoshinejad, Waleed Marei, Geraldine Hartshorne, Ali Fouladi-Nashta
    Abstract


    RD14255  Accepted 19 September 2014
    Neonatal testicular cell transplantation restores murine spermatogenesis damaged in the course of herpes simplex virus-induced orchitis
    Ekaterina Malolina, Andrey Kulibin, Alla Kushch
    Abstract


    RD14322  Accepted 16 September 2014
    Proteome profiling of the sperm maturation milieu in the rhesus monkey (Macaca mulatta) epididymis
    Xin Liu, ShaoHua Jin, XueXia Liu, WenJuan Wang, FuJun Liu
    Abstract


    RD14276  Accepted 16 September 2014
    Association of decreased spermatozoa omega-3 fatty acid levels and increased oxidative DNA damage with varicocele in infertile men: a case control study
    Li Tang, Dong Yuan, Qi Wang, Fang Jiang, Jian Guo, Yun Tang, Li Zheng, Jing Kang
    Abstract


    RD14201  Accepted 17 September 2014
    CXADR-like membrane protein (CLMP) in the rat ovary: stimulation by human chorionic gonadotropin during the periovulatory period
    Feixue Li, Xiaoping Miao, Yonglong Chen, Thomas Curry, Jr.
    Abstract


    RD14182  Accepted 17 September 2014
    N-acetylcysteine improves oocyte and embryo quality in PCOS patients undergoing ICSI: An alternative to Metformin
    Ebrahim Cheraghi, malek Soleimani Mehranjani, Mohammad Ali Shariatzadeh, Mohammad Nasr Esfahani, Zahra Ebrahimi
    Abstract


    RD14120  Accepted 16 September 2014
    Influences of melatonin treatment, MTNR1A and KiSS-1 genes polymorphisms on first conception in Sarda ewe lambs
    Sebastiano Luridiana, Maria Consuelo Mura, Cinzia Daga, Giovanni Cosso, Sara Bodano, Federico Farci, Francesca Zidda, Vincenzo Carcangiu
    Abstract


    RD14339  Accepted 11 September 2014
    TallyHO obese female mice experience poor reproductive outcomes and abnormal blastocyst metabolism which is reversed by metformin
    Kelle Moley
    Abstract


    RD14338  Accepted 11 September 2014
    Exploring the function of long non-coding RNA in the development of bovine early embryos
    Claude Robert
    Abstract


    RD14090  Accepted 12 September 2014
    Aberrant expression of TAR DNA binding protein-43 is associated with spermatogenic disorders in human males
    Divya Varghese, Uma Chandran, A Soumya, Sathy Pillai, K Jayakrishnan, Prabhakara Reddi, Pradeep Kumar
    Abstract


    RD14301  Accepted 10 September 2014
    Leukotriene production profiles and actions in the bovine endometrium during the estrous cycle
    Anna Korzekwa, Robert Milewski, Martyna Łupicka, Dariusz SkarżyÅ„ski
    Abstract


    RD14212  Accepted 11 September 2014
    Regulation of primordial follicle recruitment by cross-talking of Notch and PTEN-Akt pathways
    Linqing Wang, Jingcai Liu, Chunlei Chen, Shun-Feng Cheng, Xiaofeng Sun, Yong Zhao, Shen Yin, Zhumei Hou, Bo Pan, Cheng Ding, Wei Shen, Xifeng Zhang
    Abstract


    RD13356  Accepted 11 September 2014
    Inhibition of angiopoietin-1 (ANGPT1) affects vascular integrity in ovarian hyperstimulation syndrome (OHSS)
    Leopoldina Scotti, Dalhia Abramovich, Natalia Pascuali, Luis Haro Durand, Griselda Irusta, Ignacio de Zúñiga, Marta Tesone, fernanda parborell
    Abstract


    RD14076  Accepted 08 September 2014
    Endothelin as a local regulating factor in the bovine oviduct
    Yuki Yamamoto, Misa Kohka, Yoshihiko Kobayashi, Izabela WocÅ‚awek-Potocka, Kiyoshi Okuda
    Abstract


    RD14237  Accepted 05 September 2014
    Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality
    Noelia Prieto-Martínez, Ingrid Vilagran, Roser Morató, Joan Rodriguez-Gil, Marc Yeste, Sergi Bonet
    Abstract


    RD14140  Accepted 03 September 2014
    Ovarian acyclicity in zoo African elephants (Loxodonta africana) is associated with high body condition scores and elevated serum insulin and leptin
    Kari Morfeld, Janine Brown
    Abstract


    RD14194  Accepted 31 August 2014
    Long term cultured and cryopreserved primordial germ cells from various chicken breeds retain high proliferative potential and gonadal colonization competency
    Céline Tonus, Karine Cloquette, Fabien Ectors, Joëlle Piret, Laurent Gillet, Nadine Antoine, Daniel Desmecht, Alain Vanderplasschen, Luc Grobet, Olivier Waroux
    Abstract


    RD14317  Accepted 28 August 2014
    Gamete derivation from ESCs, iPSCs or SCNT-ESCs: State of the Art
    Gerald Schatten
    Abstract


    RD14316  Accepted 28 August 2014
    Reproductive performances: At the crossroad of genetics, technologies and environment
    Daniel Vaiman
    Abstract


    RD14311  Accepted 25 August 2014
    Genetic variation in resistance of the preimplantation bovine embryo to heat shock
    Peter Hansen
    Abstract


    RD14309  Accepted 25 August 2014
    Epigenetics, embryo quality and developmental potential
    Nathalie Beaujean
    Abstract


    RD14136  Accepted 26 August 2014
    Determinants of the valid measurement of global changes in 5'-methylcytosine and 5'-hydroxymethylcytosine by immunolocalization in the early embryo
    Juliette Salvaing, Yan Li, Nathalie Beaujean, Chris O'Neill
    Abstract


    RD14305  Accepted 22 August 2014
    Genes and environmental factors that influence disease resistance to microbes in the female reproductive tract of dairy cattle
    Martin Sheldon
    Abstract


    RD14130  Accepted 15 August 2014
    Leptin and leptin receptor are detectable in equine spermatozoa but are not involved in in vitro fertilization
    Anna Lange-Consiglio, Bruna Corradetti, Claudia Perrini, Davide Bizzaro, Fausto Cremonesi
    Abstract


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




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 5 December 2013
Parental diet, pregnancy outcomes and offspring health: metabolic determinants in developing oocytes and embryos

Kevin D. Sinclair and Adam J. Watkins

2. Published 5 December 2013
IETS 2014 abstracts


3. Published 5 December 2013
Role of stem cells in large animal genetic engineering in the TALENs–CRISPR era

Ki-Eun Park and Bhanu Prakash V. L. Telugu

4. Published 5 December 2013
Reproductive technologies and genomic selection in dairy cattle

C. Ponsart, D. Le Bourhis, H. Knijn, S. Fritz, C. Guyader-Joly, T. Otter, S. Lacaze, F. Charreaux, L. Schibler, D. Dupassieux and E. Mullaart

5. Published 5 December 2013
Genetic control of reproduction in dairy cows

Stephen T. Butler

6. Published 5 December 2013
Molecular signatures of bovine embryo developmental competence

M. Hoelker, E. Held, D. Salilew-Wondim, K. Schellander and D. Tesfaye

7. Published 5 December 2013
Editing livestock genomes with site-specific nucleases

Daniel F. Carlson, Wenfang Tan, Perry B. Hackett and Scott C. Fahrenkrug

8. Published 10 October 2013
Effect of varying glucose and glucosamine concentration in vitro on mouse oocyte maturation and developmental competence

L. A. Frank, M. L. Sutton-McDowall, D. L. Russell, X. Wang, D. K. Feil, R. B. Gilchrist and J. G. Thompson

9. Published 5 December 2013
Directional freezing of spermatozoa and embryos

Amir Arav and Joseph Saragusty

10. Published 19 June 2014
Replacing serum in culture medium with albumin and insulin, transferrin and selenium is the key to successful bovine embryo development in individual culture

E. Wydooghe, S. Heras, J. Dewulf, S. Piepers, E. Van den Abbeel, P. De Sutter, L. Vandaele and A. Van Soom

11. Published 6 January 2014
Predictive value of bovine follicular components as markers of oocyte developmental potential

Satoko Matoba, Katrin Bender, Alan G. Fahey, Solomon Mamo, Lorraine Brennan, Patrick Lonergan and Trudee Fair

12. Published 10 October 2013
Successful cloning of coyotes through interspecies somatic cell nuclear transfer using domestic dog oocytes

Insung Hwang, Yeon Woo Jeong, Joung Joo Kim, Hyo Jeong Lee, Mina Kang, Kang Bae Park, Jung Hwan Park, Yeun Wook Kim, Woo Tae Kim, Taeyoung Shin, Sang Hwan Hyun, Eui-Bae Jeung and Woo Suk Hwang

13. Published 6 January 2014
Paradoxical effect of supplementary progesterone between Day 3 and Day 7 on corpus luteum function and conceptus development in cattle

L. O'Hara, N. Forde, F. Carter, D. Rizos, V. Maillo, A. D. Ealy, A. K. Kelly, P. Rodriguez, N. Isaka, A. C. O. Evans and P. Lonergan

14. Published 12 March 2014
Effect of hCG administration during corpus luteum establishment on subsequent corpus luteum development and circulating progesterone concentrations in beef heifers

V. Maillo, P. Duffy, L. O'Hara, C. de Frutos, A. K. Kelly, P. Lonergan and D. Rizos

15. Published 5 December 2013
Embryotropic actions of follistatin: paracrine and autocrine mediators of oocyte competence and embryo developmental progression

Sandeep K. Rajput, KyungBon Lee, Guo Zhenhua, Liu Di, Joseph K. Folger and George W. Smith

16. Published 22 August 2014
Three-dimensional systems for in vitro follicular culture: overview of alginate-based matrices

Ivina R. Brito, Isadora M. T. Lima, Min Xu, Lonnie D. Shea, Teresa K. Woodruff and José R. Figueiredo

17. Published 5 December 2013
Mammalian fertility preservation through cryobiology: value of classical comparative studies and the need for new preservation options

Pierre Comizzoli and David E. Wildt

18. Published 6 January 2014
Reductions in the number of mid-sized antral follicles are associated with markers of premature ovarian senescence in dairy cows

Silvia C. Modina, Irene Tessaro, Valentina Lodde, Federica Franciosi, Davide Corbani and Alberto M. Luciano

19. Published 8 April 2014
Elements of functional genital asymmetry in the cow

B. Trigal, C. Díez, M. Muñoz, J. N. Caamaño, F. Goyache, E. Correia-Alvarez, F. J. Corrales, M. I. Mora, S. Carrocera, D. Martin and E. Gómez

20. Published 5 December 2013
Ultrasound biomicroscopy: a non-invasive approach for in vivo evaluation of oocytes and small antral follicles in mammals

L. F. M. Pfeifer, G. P. Adams, R. A. Pierson and J. Singh


      
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