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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: Graeme Martin

 
 
 

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Published online 24 August 2016
Effects of fetal calf serum on cGMP pathway and oocyte lipid metabolism in vitro 
Kátia R. L. Schwarz, Ramon Cesar Botigelli, Maite Del Collado, Fernanda Cavallari de Castro, Hugo Fernandes, Daniela M. Paschoal and Cláudia Lima Verde Leal

Serum affects lipid metabolism in oocytes and embryos, and cGMP has been shown to affect lipolysis in adipocytes. The present study shows that serum in the maturation medium interferes with lipid metabolism in oocytes and cumulus cells and these effects depend, at least in part, on the cGMP pathway. Evidence of involvement of this pathway in lipolysis in cumulus–oocyte complexes could contribute to the development of better culture systems for the in vitro production of oocytes and embryos with lower lipid accumulation and better cryotolerance.

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Published online 22 August 2016
Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations 
Rocío Fernández-Gago, Manuel Álvarez-Rodríguez, Marta E. Alonso, J. Ramiro González, Beatriz Alegre, Juan C. Domínguez and Felipe Martínez-Pastor

Seminal plasma could benefit boar semen when added during thawing. This study shows that that the addition of 10% or 50% seminal plasma during post-thawing incubation improved total motility and the motility pattern (motile subpopulations) of spermatozoa. Chromatin changes were almost non-existent for samples from some male boars, both after thawing and during the incubation, but in males susceptible to these sorts of chromatin changes, seminal plasma reduced or abolished them.

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Published online 18 August 2016
Periconceptional undernutrition affects in utero methyltransferase expression and steroid hormone concentrations in uterine flushings and blood plasma during the peri-implantation period in domestic pigs 
A. Franczak, K. Zglejc, E. Waszkiewicz, B. Wojciechowicz, M. Martyniak, W. Sobotka, S. Okrasa and G. Kotwica

A restricted diet applied to females during the peri-conceptional period may have consequences during the peri-implantation period. In this study we examined DNA methyltransferase 1 (DNMT1) and DNMT3a expression in endometrium and myometrium and oestradiol 17β (E2) and progesterone concentrations in the uterus and blood plasma in pigs during the peri-implantation period. Undernutrition during the peri-conceptional period affects maintenance and de novo DNA methylation in the endometrium, de novo DNA methylation in the myometrium and intrauterine E2 concentrations during the peri-implantation period.

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Published online 11 August 2016
Changes in testicular function proteins and sperm acrosome status in rats treated with valproic acid 
Wannisa Sukhorum and Sitthichai Iamsaard

The mechanism by which valproic acid (VPA) causes male infertility is not known. In the present study, testicular proteins and acrosome status were investigated in VPA-treated rats. Acrosome reacted spermatozoa and the expression of Ki-67, cholesterol side-chain cleavage enzyme (P450scc) and tyrosine phosphorylated proteins, except for steroidogenic acute regulatory protein, were decreased in VPA-treated rats. These changes to testicular proteins responsible for spermatogenesis result in male infertility after VPA treatment.

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Published online 10 August 2016
Potential risks to offspring of intrauterine exposure to maternal age-related obstetric complications 
Juan J. Tarín, Miguel A. García-Pérez and Antonio Cano

Several hypotheses have been proposed to explain the negative effects of delayed motherhood on an offspring’s morbidity later in life, but they are not supported by clinical and epidemiological evidence. In the present study, we show that the negative effects on offspring of delayed motherhood may be induced by embryo or fetal epigenetic DNA reprogramming associated with maternal age-related obstetric complications. This outcome has direct clinical implications for those women wanting to enrol in a fertility preservation program to offset age-related declines in fertility.

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Published online 04 August 2016
Evidence of proteinuria, but no other characteristics of pre-eclampsia, in relaxin-deficient mice 
Kelly P. O'Sullivan, Sarah A. Marshall, Scott Cullen, Tahnee Saunders, Natalie J. Hannan, Sevvandi N. Senadheera and Laura J. Parry

This paper reports that pregnant mice deficient in relaxin (Rln–/–) did not exhibit characteristics associated with preeclampsia; plasma sFlt-1 concentrations and blood pressure in pregnant Rln–/– mice were no different compared with age-matched wildtype mice. Although there was evidence of proteinuria in older pregnant Rln–/– mice, this was also observed in non-pregnant mice and was not a consequence of pregnancy. Therefore, pregnant Rln–/– mice are not a suitable animal model to study preeclampsia.

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Published online 03 August 2016
Circumventing the natural, frequent oestrogen waves of the female cheetah (Acinonyx jubatus) using oral progestin (Altrenogest) 
Adrienne E. Crosier, Pierre Comizzoli, Diana C. Koester and David E. Wildt

We found that cheetahs do not display typical estrous cycles, but rather frequent waves of estrogenic activity every 7 to 10 days. For effective ovulation induction and the most mature oocytes, gonadotropin treatments had to be given at least 3 days after an estrogen peak. Follicular activity can be suppressed by progestin supplementation, thereby providing a way to quiesce the ovary beforehand.

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Published online 01 August 2016
Beneficial effects of diazepin-quinazolin-amine derivative (BIX-01294) on preimplantation development and molecular characteristics of cloned mouse embryos 
Yanfang Huang, Xiaohong Jiang, Miao Yu, Rongfu Huang, Jianfeng Yao, Ming Li, Fangfang Zheng and Xiaoyu Yang

Somatic cell nuclear transfer is frequently associated with abnormal epigenetic modifications. The present study investigated the effects of BIX-01294 on cloned mouse preimplantation embryos and the results indicated that BIX-01294 treatment of cloned embryos has beneficial effects in terms of correcting abnormal epigenetic modifications. It will be investigated further whether BIX may affect postimplantation development by correcting epigenetic modifications.

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Published online 28 July 2016
Expression of macrophage migration inhibitory factor (MIF) in bovine oviducts is higher in the postovulatory phase than during the oestrus and luteal phase 
Asrafun Nahar and Hiroya Kadokawa

Macrophage migration inhibitory factor (MIF) may be the molecular link between maternal obesity and the quality of early embryos. This study revealed that MIF expression in the oviduct, where mammalian embryos spend their first 3 days, is higher during the postovulatory phase than during oestrus and the luteal phase in heifers. Therefore, oviduct MIF may be essential for producing good-quality embryos.

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Published online 26 July 2016
Stathmin 1 plays a role in endometrial decidualisation by regulating hypoxia inducible factor-1? and vascular endothelial growth factor during embryo implantation 
Jinhai Gou, Jia Jia, Juntao Feng, Xia Zhao, Tao Yi, Tao Cui and Zhengyu Li

The aim of this study was to explore the mechanism of action of stathmin 1 (Stmn1), a protein identified by two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) in a previous proteomic analysis, in regulating embryo implantation, a critical process in pregnancy. Stmn1 was found to play a role in decidualisation and embryo implantation by inhibiting the expression of phosphorylated Akt, hypoxia-inducible factor-1α and vascular endothelial growth factor. These findings contribute to a better understanding of the potential role of Stmn1 during embryo implantation.

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Published online 26 July 2016
Role of the Na+/K+-ATPase ion pump in male reproduction and embryo development 
D. R. Câmara, J. P. Kastelic and J. C. Thundathil

This review describes the involvement of the sodium pump Na+/K+-ATPase in various aspects of animal reproduction, including critical roles in embryonic development and specific sperm functions. In addition to regulating ion concentrations, Na+/K+-ATPase participates in several cell signalling pathways. Understanding the role of this molecule creates new opportunities to enhance or suppress various aspects of reproductive biology.

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Published online 21 July 2016
Methylated oligonucleotide (MON)-induced promoter hypermethylation is associated with repression of CDH1 expression and contributes to the migration and invasion of human trophoblast cell lines 
Xi Lan, Li-Juan Fu, Zhuo-Ying Hu, Qian Feng, Xue-Qing Liu, Xue Zhang, Xue-Mei Chen, Jun-Lin He, Ying-Xiong Wang and Yu-Bin Ding

Aberrant DNA methylation has been linked to deficiencies in trophoblast motility and invasion in pregnancy complications. We increased trophoblast cell motility and invasiveness through inducing site-specific hypermethylation and repressing gene expression of CDH1 by sense-methylated oligonucleotides (MONs). The results indicate that MON-mediated DNA methylation on the CDH1 promoter may alter gene expression and contribute to trophoblast motility and invasion.

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Published online 04 July 2016
In vitro characterisation of fresh and frozen sex-sorted bull spermatozoa 
Shauna A. Holden, Craig Murphy, Juan F. Moreno, Stephen T. Butler, Andrew R. Cromie, Patrick Lonergan and Sean Fair

The use of sex-sorted fresh spermatozoa has the potential to yield higher pregnancy rates than sex-sorted frozen spermatozoa. This study sought to characterise the in vitro quality of X and Y sex-sorted bull spermatozoa and identified the need for tailored diluents for fresh sex-sorted spermatozoa to avoid head-to-head agglutination. The use of fresh sex-sorted spermatozoa, when managed correctly, has enormous potential in the dairy industry.

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Published online 01 July 2016
Thiols of flagellar proteins are essential for progressive motility in human spermatozoa 
María Eugenia Cabrillana, María de los Ángeles Monclus, Tania Estefania Sáez Lancellotti, Paola Vanina Boarelli, Amanda Edith Vincenti, Miguel Matias Fornés, Eduardo Alfredo Sanabria and Miguel Walter Fornés

This work demonstrates that it is necessary for progressive sperm motility that flagellar proteins, like ODF1, maintain reduced cysteines after epididymal transit. To determine how oxidative changes in flagellar proteins affect sperm motility, a sulfhydryl blocker was used; sperm progressive motility was abolished without affecting the level of ATP. Incomplete oxidation in flagellar proteins of asthenozoospermic samples was also observed. Inadequate maturation or an unbalanced thiol status leads to non-progressive motility.

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Published online 27 June 2016
GRIM-19, a gene associated with retinoid-interferon-induced mortality, affects endometrial receptivity and embryo implantation 
Yang Yang, Yanyan Sun, Laiyang Cheng, Anna Li, Yanjun Shen, Ligang Jiang, Xiaohui Deng and Lan Chao

We present a new hypothesis of the molecular mechanisms responsible for implantation, which could provide an important basis for further understanding of embryo implantation. Our study shows that spontaneous elimination of GRIM-19 is required for endometrial receptivity and the embryo implantation process. Our results demonstrate the involvement of GRIM-19 in the embryo implantation process by regulating adhesion, apoptosis and immune tolerance.

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Published online 22 June 2016
Reproductive competency and mitochondrial variation in aged Syrian hamster oocytes 
Fang Li, Frank J. Castora, Wentia Ford, Khalid Alarid, Howard W. Jones and R. James Swanson

Using a hamster model, we found significant changes in mitochondrial number, structure and function as well as fecundity, ova production and follicular number in aged compared to younger animals. Old hamsters had decreased fertility rates, smaller litters, and decreased numbers of preantral follicles and MII oocytes. In addition, old ova contained large areas of collapsed, non-luminal cytoplasmic lamellae. Finally, old hamster ova contained higher levels of reactive oxygen species, lower mitochondrial membrane potential and decreased mitochondrial numbers, all of which could be contributing to the age-related decline in mammalian fertility rates.

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Published online 22 June 2016
Subfertility in androgen-insensitive female mice is rescued by transgenic FSH 
K. A. Walters, M. C. Edwards, M. Jimenez, D. J. Handelsman and C. M. Allan

Androgens (male steroids) interact with the key regulator of female reproduction, FSH, but the nature of their interaction is not clear. Examination of mutant androgen-insensitive mice with and without the expression of human transgenic FSH (Tg-FSH) revealed that subfertility in androgen-insensitive mice is rescued by the presence of Tg-FSH. These findings demonstrate that androgens and FSH actions interact in a synergistic manner to maintain optimal female fertility.

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Published online 20 June 2016
Putrescine supplementation during in vitro maturation of aged mouse oocytes improves the quality of blastocysts 
Dandan Liu, Guolong Mo, Yong Tao, Hongmei Wang and X. Johné Liu

Poor egg quality underlies reproductive aging in mice and in women. Oocyte maturation is a critical stage in determining the quality of eggs and hence embryonic potential. We demonstrated that putrescine supplementation during in vitro maturation of aged mouse oocytes resulted in healthier embryos. Peri-ovulatory putrescine supplementation may be an effective therapy in reproductive aging.

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Published online 16 June 2016
Effects of intracerebroventricular infusions of ghrelin on secretion of follicle-stimulating hormone in peripubertal female sheep 
Anna Wójcik-Gładysz, Marta Wańkowska, Alina Gajewska, Tomasz Misztal, Marlena Zielińska-Górska, Michał Szlis and Jolanta Polkowska

Negative energy balance is the limiting factor for reproduction and coordination of metabolic signals in the brain is critical during female puberty. This research focused on the effects of ghrelin, a modulator of energy balance, on reproductive activity and revealed that it modulates folliculotropin secretion in peripubertal female sheep. Ghrelin belongs to the neuroendocrine network involved in the regulation of reproductive axis activity at the brain level.

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Published online 09 June 2016
Stem cell factor promotes in vitro ovarian follicle development in the domestic cat by upregulating c-kit mRNA expression and stimulating the phosphatidylinositol 3-kinase/AKT pathway 
Paweena Thuwanut, Pierre Comizzoli, David E. Wildt, Carol L. Keefer and Nucharin Songsasen

Stem cell factor (SCF) and epidermal growth factor (EGF) play key roles in ovarian folliculogenesis. Herein we report that SCF promotes cat follicle growth by upregulating c-kit mRNA expression and AKT phosphorylation. EGF suppresses the stimulating effect of SCF, leading to a downregulation of FSH receptor mRNA expression. The ability to grow premature follicles in vitro could become a pivotal tool for preserving fertility in animal disease models and endangered felids.

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Published online 09 June 2016
Iloprost supports early development of in vitro-produced porcine embryos through activation of the phosphatidylinositol 3-kinase/AKT signalling pathway 
Pil-Soo Jeong, Seung-Bin Yoon, Seon-A Choi, Bong-Seok Song, Ji-Su Kim, Bo-Woong Sim, Young-Ho Park, Hae-Jun Yang, Seong-Eun Mun, Young-Hyun Kim, Philyong Kang, Kang-Jin Jeong, Youngjeon Lee, Yeung Bae Jin, Jae-Won Huh, Sang-Rae Lee, Deog-Bon Koo, Young Il Park, Sun-Uk Kim and Kyu-Tae Chang

Despite evidence of the presence of PGI2 in mammalian oviducts, its role in development of early embryos is largely unknown. We revealed that supplementation of PGI2 analog enhanced early development of IVP porcine embryos through activation of AKT cascade. These results indicate that iloprost can be a useful IVC supplement for production of IVP early porcine embryos with high quality.

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Published online 06 June 2016
High glucose levels affect spermatogenesis: an in vitro approach 
Renata S. Tavares, Joana M. D. Portela, Maria I. Sousa, Paula C. Mota, João Ramalho-Santos and Sandra Amaral

Diabetes mellitus (DM) has been associated with male reproductive impairment but the underlying mechanisms are poorly understood owing to the diversity of factors involved in the disease. In this study we focused on the in vitro effects of high glucose levels, the primary recognised signature of DM, on spermatogenesis. We found that while mouse testicular luminal area was decreased, Sertoli cell number was increased. These findings suggest an impairment of Sertoli cell function in high-glucose conditions that may impact spermatogenesis.

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Published online 02 June 2016
Sex of co-twin affects the in vitro developmental competence of oocytes derived from 6- to 8-week-old lambs 
Jennifer M. Kelly, David O. Kleemann, Hayley McGrice, Jose A. Len, Karen L. Kind, Will H. E. J. van Wettere and Simon K. Walker

The effect of birth type (single, twin, triplet) and co-twin sex on in vitro developmental competence of oocytes from 6 to 8-week old lambs significantly increased blastocyst development rates for lambs born with a female co-twin compared with a male co-twin. This indicates a female co-twin enhancement effect on oocyte development. Selection of lambs for embryo production programs based on co-twin sex is warranted.

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Published online 01 June 2016
Role of Ca2+ in the IVM of spermatozoa from the sterlet Acipenser ruthenus 
Olga Bondarenko, Borys Dzyuba, Marek Rodina and Jacky Cosson

Spermatozoa maturation is a necessary step for the best success in artificial reproduction. This study focused on the role of calcium ions in the acquisition of testicular sturgeon sperm to be able to swim: our results show that an influx of Ca2+ ions is required for finalisation of this maturation process.

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Published online 31 May 2016
Maternal food restriction in rats of the F0 generation increases retroperitoneal fat, the number and size of adipocytes and induces periventricular astrogliosis in female F1 and male F2 generations 
A. O. Joaquim, C. P. Coelho, P. Dias Motta, L. F. Felício, E. F. Bondan, E. Teodorov, M. F. M. Martins, T. B. Kirsten, L. V. Bonamin and M. M. Bernardi

Transgenerational food restriction and overweightAdverse environmental factors during gestation, such as under- or overnutrition, may predispose individuals to certain diseases later in life by altering their fetal programming. The present study investigated whether prenatal 40% food restriction was able to induce weight, metabolic and neuroimmune disturbances in the next two generations, even if they were normally fed. We showed a transgenerational transmission effect for overweight over two generations, associated with changes in immune and inflammatory patterns, revealing the severe impact of undernutrition over generations.

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Published online 30 May 2016
Cryptic male choice: experimental evidence of sperm sex ratio and seminal fluid adjustment in relation to coital rate 
A. M. Edwards and E. Z. Cameron

Mammalian males do not produce equal numbers of X- and Y-chromosome-bearing sperm, but factors influencing the variation are unknown. We show that sperm sex ratios and ejaculate traits vary with mating rate in laboratory mice, and hypothesise that males that mate frequently are usually more attractive, and would be advantaged by producing sons that inherit their attractiveness.

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Published online 26 May 2016
Temporary undernutrition during early gestation, corpora lutea morphometrics, ovarian progesterone secretion and embryo survival in gilts 
P. Langendijk, E. G. Bouwman, T. Y. Chen, R. E. Koopmanschap and N. M. Soede

This study looked at how off-feed incidents or competition for feed can affect hormonal regulation of luteal function and early pregnancy in pigs. We found no effect of a day’s fasting on LH secretion, but progesterone secretion was reduced and fewer piglets were born at subsequent farrowing. Malnutrition in group-housed sows or in periods of hot weather can potentially reduce ovarian progesterone output and fertility.

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Published online 26 May 2016
Non-genomic action of vitamin D3 on N-methyl-D-aspartate and kainate receptor-mediated actions in juvenile gonadotrophin-releasing hormone neurons 
Pravin Bhattarai, Janardhan P. Bhattarai, Min Sun Kim and Seong Kyu Han

Vitamin D deficiency has been linked to impaired pubertal development affecting reproductive physiology. The effects of vitamin D3 were investigated on the juvenile gonadotrophin-releasing hormone (GnRH) neuron, a central regulator of the hypothalamic–pituitary–gonadal axis. Exposure to vitamin D3 suppressed the N-methyl-D-aspartate and kainate-mediated responses of juvenile GnRH neurons. We suggest that vitamin D is essential for normal and timely functioning of pubertal development.

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Published online 25 May 2016
Aquaglyceroporins 3 and 7 in bull spermatozoa: identification, localisation and their relationship with sperm cryotolerance 
Noelia Prieto-Martínez, Roser Morató, Rodrigo Muiño, Carlos O. Hidalgo, Joan E. Rodríguez-Gil, Sergi Bonet and Marc Yeste

For the first time the presence and localisation of AQP3 and AQP7 in bull spermatozoa was determined, together with their relationship to sperm cryotolerance. Our findings show that neither the relative amounts nor the localisation of these proteins was altered by cryopreservation procedures. Furthermore, the abundance of AQP7 in fresh semen was found to predict bull sperm cryotolerance.

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Published online 25 May 2016
Membrane lipid profile of in vitro-produced embryos is affected by vitrification but not by long-term dietary supplementation of polyunsaturated fatty acids for oocyte donor beef heifers 
Beatriz C. S. Leão, Nathália A. S. Rocha-Frigoni, Ériklis Nogueira, Elaine C. Cabral, Christina R. Ferreira, Marcos N. Eberlin, Mônica F. Accorsi, Thiago V. Neves and Gisele Z. Mingoti

In this study we evaluated the effect of feeding oocyte donor beef heifers with a protected fat enriched in linoleic acid on embryonic developmental rates after IVF, embryo cryotolerance and the membrane lipid composition of fresh and vitrified–warmed embryos. Although embryo yield, cryotolerance and membrane lipid profile were unaffected by dietary supplementation, the membrane lipid profile was affected by the cryopreservation process. Because the lipid ion abundances of fresh and vitrified–warmed embryos were distinct, they can be used as potential markers of post-cryopreservation embryonic survival.

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Published online 23 May 2016
The effects of sildenafil citrate on feto–placental development and haemodynamics in a rabbit model of intrauterine growth restriction 
Jorge López-Tello, María Arias-Álvarez, Maria-Ángeles Jiménez-Martínez, Alicia Barbero-Fernández, Rosa María García-García, María Rodríguez, Pedro L. Lorenzo, Laura Torres-Rovira, Susana Astiz, Antonio González-Bulnes and Pilar G. Rebollar

Current treatments for intrauterine growth restriction are not particularly effective. Sildenafil citrate (SC), a vasodilator and angiogenic molecule, could be an option to enhance conceptus growth. Here we studied, in a rabbit model, whether the administration of SC could improve or ameliorate diet-induced defects in conceptus development and offspring outcome. SC seems to benefit pregnancies complicated by placental insufficiency; however, further research is needed.

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Published online 23 May 2016
Variation among individual bulls in the distribution of acrosomal tyrosine-phosphorylated proteins in epididymal and ejaculated spermatozoa 
Miyuki M. Arai, Kenta Minami, Yukari Ogura, Nagisa Otsuka, Shohei Hama, Hiroshi Harayama, Mitsuhiro Sakase and Moriyuki Fukushima

A valid biomarker for sperm molecular characterisation was found in Japanese black cattle. The distribution of acrosomal tyrosine-phosphorylated proteins largely differs among individual bulls. The grades of acrosomal freezability and results of AI with cryopreserved spermatozoa can be predicted by assessing the distribution of acrosomal tyrosine-phosphorylated proteins in spermatozoa before cryopreservation.

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Published online 17 May 2016
Expressions of lipoprotein receptors and cholesterol efflux regulatory proteins during luteolysis in bovine corpus luteum 
Kei Horihata, Shin Yoshioka, Masahiro Sano, Yuki Yamamoto, Koji Kimura, Dariusz J. Skarzynski and Kiyoshi Okuda

Progesterone, which is essential for pregnancy in mammals, is synthesised from cholesterol in the corpus luteum (CL). We investigated the involvement of cholesterol uptake receptors and efflux transporters in bovine CL regression (luteolysis), and found that the expression of scavenger receptor B1, a cholesterol uptake receptor, decreased during luteolysis. The decline in cholesterol uptake may reduce progesterone production in the CL during luteolysis.

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Published online 17 May 2016
The effects in vitro of TNF-α and its antagonist ‘etanercept’ on ejaculated human sperm 
Nicola A. Pascarelli, Antonella Fioravanti, Elena Moretti, Giacomo M. Guidelli, Lucia Mazzi and Giulia Collodel

The effect of cytokines on human male fertility is attracting more attention. In this in vitro study, the effects of TNF-α and its antagonist, etanercept, in ejaculated human spermatozoa were analysed. The negative influence of TNF-α on sperm function and the positive action of etanercept were demonstrated. This data may have clinical relevance for the management of chronic inflammatory diseases related to male reproduction.

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Published online 17 May 2016
Effects of fetal genotype and sex on developmental response to maternal malnutrition 
Laura Cogollos, Consolacion Garcia-Contreras, Marta Vazquez-Gomez, Susana Astiz, Raul Sanchez-Sanchez, Ernesto Gomez-Fidalgo, Cristina Ovilo, Beatriz Isabel and Antonio Gonzalez-Bulnes

The occurrence of intrauterine growth restriction is linked to a plethora of non-communicable disorders during postnatal life. Our study, using a translational swine model, provides evidence that prenatal developmental is strongly modulated by fetal genotype and sex. Such findings stress the necessity for considering ethnicity and sex effects in the study of the developmental origins of adult health and disease.

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Published online 17 May 2016
Actions of activin A, connective tissue growth factor, hepatocyte growth factor and teratocarcinoma-derived growth factor 1 on the development of the bovine preimplantation embryo 
Jasmine Kannampuzha-Francis, Paula Tribulo and Peter J. Hansen

The reproductive tract secretes molecules collectively known as embryokines that regulate embryonic growth and development. In this study we tested four growth factors expressed in the endometrium for actions on the bovine preimplantation embryo. All four molecules affected embryonic development or gene expression and may therefore function in the endometrium as embryokines.

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Published online 12 May 2016
Temporal regulation of extracellular signal-regulated kinase 1/2 phosphorylation, heat shock protein 70 and activating transcription factor 3 during prostaglandin F-induced luteal regression in pseudopregnant rats following heat stress 
Wu-jiao Bai, Peng-jing Jin, Mei-qian Kuang, Quan-wei Wei, Fang-xiong Shi, John S. Davis and Da-gan Mao

High temperatures are correlated with reduced fertility. Ovarian function in pseudopregnant rats was determined in a laboratory setting of repeated heat stress. Heat stress altered the activity of mitogen-activated protein kinases and the expression of heat shock protein 70 and activating transcription factor 3 in the corpus luteum. A deeper understanding of the effects of heat stress on ovarian function will enable better approaches to promote female fecundity and sustainable animal production.

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Published online 11 May 2016
Roles of interferon-stimulated gene 15 protein in bovine embryo development 
Shuan Zhao, Yi Wu, Hui Gao, Alexander Evans and Shen-Ming Zeng

ISG15 mRNA exists in bovine embryo from one cell stage, which is earlier than IFNT mRNA. When the expression of ISG15 was knockdown, the development of blastocyst was negatively affected, , and the expressions of Ets2, IFNT and connexion 43 were inhibited in bovine blastocysts.

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Published online 11 May 2016
p21-activated kinase 1 activity is required for histone H3 Ser10 phosphorylation and chromatin condensation in mouse oocyte meiosis 
Nana Zhang, Xiuhong Li, Xiaoyun Liu, Yan Cao, Dandan Chen, Xiaoyu Liu, Qian Wang, Juan Du, Jing Weng and Wei Ma

Histone H3 phosphorylation is essential for chromatin condensation, which is a prerequisite for accurate chromosome separation in mitosis or meiosis. The present study demonstrates that p21-activated kinase 1 (Pak1) activity is required for histone H3 phosphorylation and consequent chromatin condensation in mouse oocytes upon the resumption of meiotic progression.

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Published online 11 May 2016
Implantation rates subsequent to the transfer of embryos produced at different phases during double stimulation of poor ovarian responders 
Qiong Zhang, Xiaoyue M. Guo and Yanping Li

Feasibility of luteal phase ovarian stimulation has already be certified in poor ovarian responders. Embryos produced during the luteal phase resulted in higher implantation rates compared with those produced during the follicular phase. Luteal phase ovarian stimulation should be recommended for poor ovarian responders.

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Published online 11 May 2016
Perinatal exposure to insecticide fipronil: effects on the reproductive system in male rats 
Aline L. de Barros, Julie H. Bae, Cibele S. Borges, Josiane L. Rosa, Marilia M. Cavariani, Patrícia V. Silva, Patricia F. F. Pinheiro, Janete A. Anselmo-Franci and Arielle C. Arena

Fipronil, an insecticide widely used in agriculture, veterinary medicine and public health, has been listed as a potential endocrine disrupter. The present study showed that fipronil can affect the sperm motility and compromise sperm quality of the animals. The perinatal exposure to fipronil has long-term effects on sperm parameters, and the epididymis can be a target organ.

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    | Supplementary Material (196 KB)
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Published online 11 May 2016
Identification of motility-associated progesterone-responsive differentially phosphorylated proteins 
V. Sagare-Patil and D. Modi

Progesterone is a steroid present in the female reproductive tract that aids sperm motility. In the present study we identified human sperm proteins that are modified in response to progesterone and propose a mechanism by which progesterone would aid motility. This information may be useful for the development of drugs to improve sperm motility and contraceptives.

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    | Supplementary Material (47 KB)
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Published online 11 May 2016
Prominin-1 glycosylation changes throughout early pregnancy in uterine epithelial cells under the influence of maternal ovarian hormones 
Samson N. Dowland, Romanthi J. Madawala, Connie E. Poon, Laura A. Lindsay and Christopher R. Murphy

The prominin-1 protein has previously been broadly investigated for its potential role in cancer and stem cell biology. This is the first investigation into the role of prominin-1 in early pregnancy. The study localises prominin-1 to the apical region of uterine epithelial cells and shows that maternal ovarian hormones significantly affect the glycosylation and abundance of this protein.

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Published online 11 May 2016
Anti-Müllerian hormone reduces growth rate without altering follicular survival in isolated caprine preantral follicles cultured in vitro 
R. M. P. Rocha, L. F. Lima, I. R. Brito, G. M. Silva, H. H. V. Correia, N. A. Ribeiro de Sá, A. C. A. Ferreira, A. D. Sales, C. H. Lobo, C. C. Campello, J. Smitz, M. B. Wheeler and J. R. Figueiredo

Investigations into the relationship between FSH and anti-Müllerian hormone (AMH) during in vitro culture of isolated preantral follicles are lacking. The aim of the present study was to evaluate the effect of AMH, with and without FSH, on the in vitro development of isolated caprine follicles. We demonstrated that AMH reduces follicular growth without altering follicular survival. Thus, AMH may play an important role in regulating the growth of caprine preantral follicles.

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Published online 10 May 2016
Variable duration of reproductive suppression in male coyotes (Canis latrans) treated with a high dose of the gonadotrophin-releasing hormone agonist deslorelin 
Marjorie J. MacGregor, Cheryl S. Asa and Donal C. Skinner

A unique approach to managing coyote (Canis latrans) depredation is the development of a one-time deliverable chemical sterilant. We hypothesised that exposure to a high dose of a gonadotrophin-releasing hormone (GnRH) agonist would cause prolonged suppression of the reproductive axis. The results of the present study suggest that a plasma concentration of 100 pg mL–1 deslorelin must be maintained and that elevations beyond this threshold are likely to be unimportant.

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    | Supplementary Material (89 KB)
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Published online 09 May 2016
Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates 
C. M. Kershaw, G. Evans, R. Rodney and W. M. C. Maxwell

Sperm freezing has advanced animal breeding and genetics in numerous species but is not successful in alpacas due to the gel-like nature of the semen. This research aimed to reduce the viscosity of alpaca semen without affecting sperm function. The enzyme papain (0.1 mg mL–1 for 20 min at 37°C) followed by the papain inhibitor E-64 (10 µm for 5min at 37°C) eliminated viscosity and aided cryopreservation of alpaca spermatozoa. These findings will help to advance the use of reproductive technologies in camelids.

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Published online 02 May 2016
Optineurin suppression activates the mediators involved in the terminal effector pathways of human labour and delivery 
Ratana Lim, Gillian Barker and Martha Lappas

Preventing spontaneous preterm birth, which is the leading cause of perinatal mortality and morbidity, is one of the most important issues facing perinatal medicine today. In this study, we found that spontaneous labour is associated with an increase in optineurin (OPTN) expression in fetal membranes and the myometrium, where it functions as a negative regulator of prolabour mediators. Elucidating the role of OPTN in labour processes enhances our understanding of the mechanisms that drive preterm birth and could perhaps lead to the development of a clinical therapeutic strategy.

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    | Supplementary Material (77 KB)
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Published online 28 April 2016
Proteome of equine oviducal fluid: effects of ovulation and pregnancy 
Katrien Smits, Hilde Nelis, Katleen Van Steendam, Jan Govaere, Kim Roels, Cyrillus Ververs, Bart Leemans, Eline Wydooghe, Dieter Deforce and Ann Van Soom

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

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    | Supplementary Material (400 KB)
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Published online 26 April 2016
Sperm function during incubation with oestrus oviductal fluid differs in bulls with different fertility 
A. Kumaresan, A. Johannisson and A.-S. Bergqvist

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

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

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

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

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

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

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

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

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

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    | Supplementary Material (19 KB)
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Published online 06 April 2016
Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen 
J. M. Morrell, A. Lagerqvist, P. Humblot and A. Johannisson

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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   | Supplementary Material (75 KB)  |        Open Access Article
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Published online 29 February 2016
Greater sperm complexity in the Australasian old endemic rodents (Tribe: Hydromyini) is associated with increased levels of inter-male sperm competition 
Hanna J. McLennan, Stefan Lüpold, Pete Smissen, Kevin C. Rowe and William G. Breed

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

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    | Supplementary Material (154 KB)
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Published online 24 February 2016
Effect of spermatozoa motility hyperactivation factors and gamete coincubation duration on in vitro bovine embryo development using flow cytometrically sorted spermatozoa 
Luis B. Ferré, Yanina Bogliotti, James L. Chitwood, Cristóbal Fresno, Hugo H. Ortega, Michael E. Kjelland and Pablo J. Ross

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

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

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

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

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

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

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

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

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

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

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

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

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

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    | Supplementary Material (32 KB)
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Published online 05 February 2016
Preantral follicle density in ovarian biopsy fragments and effects of mare age 
K. A. Alves, B. G. Alves, G. D. A. Gastal, K. T. Haag, M. O. Gastal, J. R. Figueiredo, M. L. Gambarini and E. L. Gastal

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

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

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

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

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

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    | Supplementary Material (92 KB)
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Published online 14 January 2016
Effects of prepartum oilseed supplements on subclinical endometritis, pro- and anti-inflammatory cytokine transcripts in endometrial cells and postpartum ovarian function in dairy cows 
Reza Salehi, Marcos G. Colazo, Mohanathas Gobikrushanth, Urmila Basu and Divakar J. Ambrose

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

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

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

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    | Supplementary Material (79 KB)
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Published online 11 January 2016
Mammalian target of rapamycin/eukaryotic initiation factor 4F pathway regulates follicle growth and development of theca cells in mice 
Chao Zhang, Xiao-Ran Liu, Yong-Chun Cao, Jin-Ling Tian, Di Zhen, Xiao-Fei Luo, Xin-Mei Wang, Jian-Hui Tian and Jian-Ming Gao

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

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    | Supplementary Material (83 KB)
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Published online 06 January 2016
LIM kinase activity is required for microtubule organising centre positioning in mouse oocyte meiosis 
Xin Li, Yubo Zhu, Yan Cao, Qian Wang, Juan Du, Jianhui Tian, Yuanjing Liang and Wei Ma

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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    | Supplementary Material (38 KB)
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Published online 05 May 2015
Novel equine conceptus–endometrial interactions on Day 16 of pregnancy based on RNA sequencing 
C. Klein

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

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    | Supplementary Material (1.2 MB)
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Published online 05 May 2015
Dynamics of Notch signalling in the mouse oviduct and uterus during the oestrous cycle 
D. Murta, M. Batista, A. Trindade, E. Silva, L. Mateus, A. Duarte and L. Lopes-da-Costa

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

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    | Supplementary Material (50 KB)
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Published online 05 May 2015
Potential role of maternal lineage in the thoroughbred breeding strategy 
Xiang Lin, Shi Zhou, Li Wen, Allan Davie, Xinkui Yao, Wujun Liu and Yong Zhang

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

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    | Supplementary Material (356 KB)
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Published online 01 May 2015
Use of the gonadotrophin-releasing hormone antagonist azaline B to control the oestrous cycle in the koala (Phascolarctos cinereus) 
K. Ballantyne, S. T. Anderson, M. Pyne, V. Nicolson, A. Mucci, A. Lisle and S. D. Johnston

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

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

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

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blank image Reproduction, Fertility and Development
Volume 28 Number 10 2016

 
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Epigenetics and developmental programming of welfare and production traits in farm animals 
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K. D. Sinclair , K. M. D. Rutherford , J. M. Wallace , J. M. Brameld , R. Stöger , R. Alberio , D. Sweetman , D. S. Gardner , V. E. A. Perry , C. L. Adam , C. J. Ashworth , J. E. Robinson and C. M. Dwyer
pp. 1443-1478

Farm animals have been used extensively as models to study prenatal programming of health and disease but comparatively little attention has been directed towards traits of commercial importance until recently. We review the evidence that prenatal factors such as parental nutrition, gestational stress, exposure to environmental chemicals and advanced breeding technologies, can influence traits such as postnatal growth, feed efficiency, milk yield, carcass composition, animal welfare and reproductive potential; and discuss underlying epigenetic mechanisms of inheritance. We conclude that, although proof of concept is proven for several traits, issues relating to effect-size for complex traits that are polygenic in nature are small, and hence commercial importance remains uncertain. Resolution of these discrepancies will require concerted international efforts that consider multiple concurrent stressors that recapitulate contemporary systems of livestock production.

   | Supplementary Material (617 KB)  |        Open Access Article
 

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Multiple roles of hypoxia in ovarian function: roles of hypoxia-inducible factor-related and -unrelated signals during the luteal phase 
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Ryo Nishimura and Kiyoshi Okuda
pp. 1479-1486

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

 
  
 

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Transcriptome of early embryonic invasion at implantation sites in a murine model 
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J. M. Moreno-Moya , N. A. Franchi , S. Martínez-Escribano , J. A. Martínez-Conejero , S. Bocca , S. Oehninger and J. A. Horcajadas
pp. 1487-1498

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

 
    | Supplementary Material (105 KB)
 

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Comparative genomic identification and expression profiling of a novel β-defensin gene cluster in the equine reproductive tract 
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Gillian P. Johnson , Andrew T. Lloyd , Cliona O’Farrelly , Kieran G. Meade and Sean Fair
pp. 1499-1508

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

 
  
 

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Different locations of RANTES and its receptors on mouse epididymal spermatozoa 
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Jin-Hua Wei , Xiao Feng , Zhi-Jian Sun , Pang Cheng , Bin-Fang Ma , Jie Zhao , Yu-Hang Dong , Yuan-Qiang Zhang and Zhen Li
pp. 1509-1517

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

 
  
 

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The employment of IVF techniques for establishment of sodium, copper and selenium impact upon human sperm quality 
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Urszula Marzec-Wróblewska , Piotr Kamiński , Paweł Łakota , Marek Szymański , Karolina Wasilow , Grzegorz Ludwikowski , Magdalena Kuligowska-Prusińska , Grażyna Odrowąż-Sypniewska , Tomasz Stuczyński and Alina Woźniak
pp. 1518-1525

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

 
  
 

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The use of a synthetic progesterone, levonorgestrel (LNG), to control the oestrous cycle in the koala 
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K. Ballantyne , S. T. Anderson , M. Pyne , V. Nicolson , A. Mucci , A. Lisle and S. D. Johnston
pp. 1526-1532

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

 
  
 

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Spatio-specific regulation of endocrine-responsive gene transcription by periovulatory endocrine profiles in the bovine reproductive tract 
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Estela R. Araújo , Mariana Sponchiado , Guilherme Pugliesi , Veerle Van Hoeck , Fernando S. Mesquita , Claudia M. B. Membrive and Mario Binelli
pp. 1533-1544

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

 
  
 

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Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques 
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J. Dorado , M. J. Gálvez , S. Demyda-Peyrás , I. Ortiz , J. M. Morrell , F. Crespo , J. Gósalvez and M. Hidalgo
pp. 1545-1552

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

 
  
 

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C-Type natriuretic peptide maintains domestic cat oocytes in meiotic arrest 
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Yougang Zhong , Jiabao Lin , Xiaoping Liu , Jian Hou , Yong Zhang and Xingxu Zhao
pp. 1553-1559

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

 
  
 

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Spindle configuration and developmental competence of in vitro-matured bovine oocytes exposed to NaCl or sucrose prior to Cryotop vitrification 
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Núria Arcarons , Roser Morató , Jozé F. W. Spricigo , Marcia A. M. M. Ferraz and Teresa Mogas
pp. 1560-1569

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

 
  
 

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Hypoxia limits mouse follicle growth in vitro 
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J. M. Connolly , M. T. Kane , L. R. Quinlan , P. Dockery and A. C. Hynes
pp. 1570-1579

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

 
  
 

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MicroSort® sperm sorting causes no increase in major malformation rate 
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Donald P. Marazzo , David Karabinus , Lawrence A. Johnson and Joseph D. Schulman
pp. 1580-1587

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

   |        Open Access Article
 

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Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2α and PGE2 production in the bovine endometrium 
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Mariko Kuse , Ryosuke Sakumoto and Kiyoshi Okuda
pp. 1588-1597

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

 
  
 

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miR-424/322 is downregulated in the semen of patients with severe DNA damage and may regulate sperm DNA damage 
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Kai Zhao , Yaoping Chen , Ruifeng Yang , Yang Bai , Cuiling Li , Honggang Li and Chengliang Xiong
pp. 1598-1607

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

 
  
 

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Changes in lysosomal enzymes and mannose-6-phosphate receptors related to sexual maturation in bull epididymis 
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Andrea C. Aguilera , Lorena Carvelli , Verónica Boschin , Fabián Mohamed , Leila Zyla and Miguel A. Sosa
pp. 1608-1617

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

 
  
 

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Association of a region of bovine chromosome 1 (BTA1) with age at puberty in Angus bulls 
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María E. Fernández , Alberto Prando , Andrés Rogberg-Muñoz , Pilar Peral-García , Andrés Baldo , Guillermo Giovambattista and Juan P. Lirón
pp. 1618-1621

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

 
    | Supplementary Material (667 KB)
 

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Relationship between seminal plasma tuberoinfundibular peptide of 39 residues and sperm functional attributes in buffalo (Bubalus bubalis) 
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Sellappan Selvaraju , Lakshminarayana Somashekar , Binsila B. Krishnan , Sivashanmugam Parthipan , Guvvala Pushparani , Arunachalam Arangasamy , Duraisamy Rajendran and Janivara Parameswaraiah Ravindra
pp. 1622-1630

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

 
    | Supplementary Material (38 KB)
 

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Potential effects of interferon regulatory factor 4 in a murine model of polyinosinic-polycytidylic acid-induced embryo resorption 
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Jing Wang , Tailang Yin , Yanqi Wen , Fuju Tian , Xiaojun He , Danni Zhou , Yi Lin and Jing Yang
pp. 1631-1641

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

 
  
 

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Expression of leukaemia inhibitory factor at the conceptus–maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare 
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M. de Ruijter-Villani , C. Deelen and T. A. E. Stout
pp. 1642-1651

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

 
  
 

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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.

    RD16238  Accepted 24 August 2016
    Imprinted and DNA methyltransferase gene expression in the endometrium during the pre- and peri-implantation period in cattle.
    Alan O'Doherty, Lynne O'Shea, Olivier Sandra, Patrick Lonergan, Trudee Fair, Niamh Forde
    Abstract


    RD16164  Accepted 23 August 2016
    Testicular parameters and spermatogenesis in different birth weight boars
    Patricia Auler, Gustavo Moreira, Charis Hogg, Cheryl Ashworth, F Bortolozzo, Helio Chiarini-Garcia, Fernanda Almeida
    Abstract


    RD16269  Accepted 23 August 2016
    Sperm antioxidant defenses decrease during epididymal transit from caput to cauda in parallel with their increases in epididymal fluid in the Goat (Capra hircus)
    Mashidur Rana, Sudhir Roy, Bannur Divyashree
    Abstract


    RD15486  Accepted 19 August 2016
    Discs large homolog 1 coordinates mouse oocyte polarization during maturation
    Junchao Wang, Hong Lv, Keliang Wu, Yunshan Zhang, Hainning Luo, Zijiang Chen
    Abstract


    RD16135  Accepted 18 August 2016
    Fish oil and wheat germ oil supplementation improves ovarian dysfunction in streptozotocin-diabetic rats
    Naglaa Khedr
    Abstract


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


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


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


    RD16317  Accepted 15 August 2016
    Logistics of large scale commercial IVF embryo production
    Patrick Blondin
    Abstract


    RD16318  Accepted 15 August 2016
    Emerging role of extracellular vesicles in communication of preimplantation embryos in vitro
    Ann Van Soom
    Abstract


    RD15553  Accepted 16 August 2016
    In vivo and in vitro maturation of rabbit oocyte differently affects the gene expression profile, mitochondrial distribution, apoptosis and early embryo development
    Maria Arias-Alvarez, Rosa Maria García-García, Jorge Lopez-Tello, Pilar Rebollar, Alfonso Gutierrez-Adan, Pedro Lorenzo
    Abstract


    RD16063  Accepted 15 August 2016
    Proliferation and apoptosis processes in the seasonal testicular development of the wild Daurian ground squirrel (citellus dauricus brandt, 1844)
    Yingying Han, Jinqi Zhan, Ying Xu, Fengwei Zhang, Zhengrong Yuan, Qiang Weng
    Abstract


    RD15537  Accepted 07 August 2016
    Characterisation of Anti-Müllerian hormone in ex situ beluga (Delphinapterus leucas)
    Gisele Montano, Todd Robeck, Karen Steinman, Justine O'Brien
    Abstract


    RD16122  Accepted 03 August 2016
    A combination of IGF-1 and FSH promotes proliferation of pre-pubertal bovine Sertoli cells isolated and cultured in vitro
    Alysha Dance, John Kastelic, Jacob Thundathil
    Abstract


    RD15488  Accepted 03 August 2016
    In vitro oocyte maturation in a medium containing reduced sodium chloride improves developmental competence of pig oocytes after parthenogenesis and somatic cell nuclear transfer
    Joohyeong Lee, Hanna Lee, Yongjin Lee, Bola Park, Fazle Elahi, Seung Tae Lee, Chun-keun Park, Sang-Hwan Hyun, Eunsong Lee
    Abstract


    RD15489  Accepted 02 August 2016
    Expression of C-type lectin receptors and Toll-like receptors in decidua of patients with unexplained recurrent spontaneous abortion
    Liang Xu, Tian Qiu, Yudong Wang, Yan Chen, Weiwei Cheng
    Abstract


    RD16119  Accepted 29 July 2016
    Consumption of a High Fat Diet Alters the Seminal Fluid and Gut Microbiomes in Male Mice
    Angela Javurek, William Spollen, Sarah Johnson, Nathan Bivens, Karen Bromert, Scott Givan, Cheryl Rosenfeld
    Abstract


    RD16293  Accepted 29 July 2016
    The expression of microRNA in male reproductive tissues and their role in male fertility
    Scott Pratt
    Abstract


    RD16292  Accepted 29 July 2016
    Exploring early differentiation and pluripotency in domestic animals
    Michael Roberts
    Abstract


    RD16075  Accepted 16 July 2016
    The targeted deletion of the Kv6.4 subunit causes male sterility due to a disturbed spermiogenesis
    Glenn Regnier, Elke Bocksteins, Waleed Marei, Isabel Pintelon, Jean-Pierre Timmermans, Jo Leroy, Dirk Snyders
    Abstract


    RD16190  Accepted 04 July 2016
    Effects of freezing and activation on membrane quality and DNA damage in Xenopus tropicalis and Xenopus laevis spermatozoa
    Sean Morrow, Jaime Gosalvez, Carmen Lopez-Fernandez, Paqui Arroyo, Bill Holt, Matthew Guille
    Abstract


    RD16110  Accepted 24 June 2016
    Profiling bovine blastocyst microRNAs using deep sequencing
    Rolando Pasquariello, Beatriz Fernandez-Fuertes, Francesco Strozzi, Flavia Pizzi, Raffaele Mazza, Patrick Lonergan, Fulvio Gandolfi, John L. Williams
    Abstract


    RD15524  Accepted 24 June 2016
    Analysis of in vitro follicle development during the onset of premature ovarian insufficiency in a mouse model
    Heidy Kaune, Sairah Sheikh, Suzannah Williams
    Abstract


    RD15514  Accepted 17 May 2016
    The role of insulin-like growth factor-I on crossbred Bos indicus cattle germinal vesicle oocytes exposed to heat shock
    Rafaela Lima, Pedro Henrique Risolia, Jéssica Ispada, Mayra Assumpção, José Antonio Visintin, Cássia Orlandi, Fabíola Paula-Lopes
    Abstract


24


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Rank Paper Details
1. Published 3 December 2015
IETS 2016 abstracts


2. Published 3 December 2015
Pregnancy losses in cattle: potential for improvement

M. G. Diskin, S. M. Waters, M. H. Parr and D. A. Kenny

3. Published 3 December 2015
Causes and consequences of oxidative stress in spermatozoa

Robert John Aitken, Zamira Gibb, Mark A. Baker, Joel Drevet and Parviz Gharagozloo

4. Published 3 December 2015
Role of progesterone in embryo development in cattle

Pat Lonergan, Niamh Forde and Thomas Spencer

5. Published 3 December 2015
Mechanisms of epigenetic remodelling during preimplantation development

Pablo Juan Ross and Sebastian Canovas

6. Published 3 December 2015
Fertility and genomics: comparison of gene expression in contrasting reproductive tissues of female cattle

P. A. McGettigan, J. A. Browne, S. D. Carrington, M. A. Crowe, T. Fair, N. Forde, B. J. Loftus, A. Lohan, P. Lonergan, K. Pluta, S. Mamo, A. Murphy, J. Roche, S. W. Walsh, C. J. Creevey, B. Earley, S. Keady, D. A. Kenny, D. Matthews, M. McCabe, D. Morris, A. O'Loughlin, S. Waters, M. G. Diskin and A. C. O. Evans

7. Published 3 December 2015
Measuring embryo metabolism to predict embryo quality

Jeremy G. Thompson, Hannah M. Brown and Melanie L. Sutton-McDowall

8. Published 18 May 2016
Prospects for immunocontraception in feral horse population control: exploring novel targets for an equine fertility vaccine

Aleona Swegen and R. John Aitken

9. Published 13 January 2016
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

10. Placenta-derived extracellular vesicles: their cargo and possible functions

Mary Familari, Tina Cronqvist, Zahra Masoumi and Stefan R. Hansson

11. Effect of C-type natriuretic peptide on maturation and developmental competence of immature mouse oocytes in vitro

Qiang Wei, Chuan Zhou, Min Yuan, Yuyang Miao, Xiaoe Zhao and Baohua Ma

12. Published 8 October 2015
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

13. Follicular fluid and serum metabolites in Holstein cows are predictive of genetic merit for fertility

S. G. Moore, A. O'Gorman, L. Brennan, T. Fair and S. T. Butler

14. Published 13 January 2016
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

15. Published 3 December 2015
Breeding animals for quality products: not only genetics

Pascale Chavatte-Palmer, Anne Tarrade, Hélène Kiefer, Véronique Duranthon and Hélène Jammes

16. Published 18 April 2016
Suppressed expression of macrophage migration inhibitory factor in the oviducts of lean and obese cows

Asrafun Nahar and Hiroya Kadokawa

17. Published 21 July 2016
The impact of storage temperature and sperm number on the fertility of liquid-stored bull semen

Craig Murphy, Shauna A. Holden, Edel M. Murphy, Andrew R. Cromie, Patrick Lonergan and Sean Fair

18. Published 21 July 2016
An integrated approach to bovine oocyte quality: from phenotype to genes

Leslie Angulo, Catherine Guyader-Joly, Sylvain Auclair, Christelle Hennequet-Antier, Pascal Papillier, Mekki Boussaha, Sébastien Fritz, Karine Hugot, François Moreews, Claire Ponsart, Patrice Humblot and Rozenn Dalbies-Tran

19. Published 18 August 2016
Epigenetics and developmental programming of welfare and production traits in farm animals

K. D. Sinclair, K. M. D. Rutherford, J. M. Wallace, J. M. Brameld, R. Stöger, R. Alberio, D. Sweetman, D. S. Gardner, V. E. A. Perry, C. L. Adam, C. J. Ashworth, J. E. Robinson and C. M. Dwyer

20. Combinatorial effects of epidermal growth factor, fibroblast growth factor 2 and insulin-like growth factor 1 on trophoblast cell proliferation and embryogenesis in cattle

Ming Xie, Sarah R. McCoski, Sally E. Johnson, Michelle L. Rhoads and Alan D. Ealy


      
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