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Vertebrate reproductive science and technology

Reproduction, Fertility and Development

Reproduction, Fertility and Development

Reproduction, Fertility and Development publishes research and reviews on all aspects of vertebrate reproduction and developmental biology. Read more about the journalMore

Editor-in-Chief: Graeme Martin

Current Issue

Reproduction, Fertility and Development

Volume 29 Number 8 2017

RD16091Role of the Na+/K+-ATPase ion pump in male reproduction and embryo development

D. R. Câmara, J. P. Kastelic and J. C. Thundathil
pp. 1457-1467

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.

RD16163Potential risks to offspring of intrauterine exposure to maternal age-related obstetric complications

Juan J. Tarín, Miguel A. García-Pérez and Antonio Cano
pp. 1468-1476

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.

RD16056Evidence 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
pp. 1477-1485

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.

RD16007Circumventing 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
pp. 1486-1498

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.

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.

RD16031Methylated 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
pp. 1509-1520

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.

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.

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.

The developmental potential of follicles from a mouse model of premature ovarian insufficiency (POI) was assessed using follicle culture. Although, follicles cultured from 3- and 6-week-old mice had decreased survival compared to Controls, a population of follicles retain the potential of full development in vitro. These results indicate that in vitro follicle culture is a promising method to rescue follicle development after the onset of POI in this model.

RD16110Profiling bovine blastocyst microRNAs using deep sequencing

R. Pasquariello, B. Fernandez-Fuertes, F. Strozzi, F. Pizzi, R. Mazza, P. Lonergan, F. Gandolfi and J. L. Williams
pp. 1545-1555

MicroRNAs (miRNAs) control several reproductive functions, including oocyte maturation, implantation and early embryonic development. We developed a method for accurate miRNA identification in a small sample of bovine blastocysts. Using pools of only 30 embryos, we identified miRNAs that regulate target genes and metabolic pathways that had not been associated with early embryogenesis before, and are potential biomarkers of developmental competence.

RD16190Effects of freezing and activation on membrane quality and DNA damage in Xenopus tropicalis and Xenopus laevis spermatozoa

S. Morrow, J. Gosálvez, C. López-Fernández, F. Arroyo, W. V. Holt and M. J. Guille
pp. 1556-1566

Xenopus frogs are key organisms for studying basic biology and for modelling human disease. Genetically altered lines of frogs are therefore made, and it is important for animal welfare and cost-effectiveness to store these as frozen spermatozoa. In this study we show that Xenopus sperm cryopreservation damages the DNA and plasma membrane of spermatozoa and start to identify factors affecting damage levels.

RD16075Targeted deletion of the Kv6.4 subunit causes male sterility due to disturbed spermiogenesis

Glenn Regnier, Elke Bocksteins, Waleed F. Marei, Isabel Pintelon, Jean-Pierre Timmermans, Jo L. M. R. Leroy and Dirk J. Snyders
pp. 1567-1575

Several voltage-gated K+ (Kv) channel subunits are expressed in testis, but their contribution to testicular function remains largely unknown. We provide direct evidence of the relevance of Kv6.4 for male fertility since the targeted deletion of this gene in mice disrupted spermiogenesis, resulting in male sterility. This makes Kv6.4 a potentially interesting pharmacological target for the development of non-hormonal male contraceptives.

RD15530Thawing 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
pp. 1576-1584

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.

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.

RD15512Effects 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
pp. 1593-1601

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.

RD16119Consumption of a high-fat diet alters the seminal fluid and gut microbiomes in male mice

Angela B. Javurek, William G. Spollen, Sarah A. Johnson, Nathan J. Bivens, Karen H. Bromert, Scott A. Givan and Cheryl S. Rosenfeld
pp. 1602-1612

We recently discovered that the male seminal vesicles harbour a novel seminal fluid microbiome (SFM). The present study shows that short-term feeding of a high-fat diet (HFD) changes the bacterial composition of the seminal fluid and fecal microbiomes. HFD-induced SFM alterations might lead to detrimental health consequences in males, their reproductive partners and resulting offspring.

This study aimed to explore the mechanisms underlying the pathogenesis of unexplained recurrent spontaneous abortion (URSA). The results suggest that the balance between C-type lectin-like receptors and Toll-like receptors (TLRs) was tilted towards a TLR-dominant response in URSA patients, which may disrupt maternal–fetal immune tolerance, resulting in spontaneous abortion.

RD15488In vitro oocyte maturation in a medium containing reduced sodium chloride improves the 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, Choon-Keun Park, Sang-Hwan Hyun and Eunsong Lee
pp. 1625-1634

The quality of oocytes is a crucial factor in the field of reproductive biotechnology. We observed that maturation of pig oocytes in hypotonic medium with reduced NaCl improves the developmental competence of oocytes by affecting the cytoplasmic microenvironment. An oocyte maturation system using a medium with reduced NaCl could potentially improve the efficiency of reproduction in livestock species after further optimisation.

Sertoli cells, which support and coordinate developing sperm in the testes, were isolated from 8-week-old Holstein bull calves. A combination of two hormones (IGF-I and FSH) enhanced multiplication of Sertoli cells. We concluded that increased IGF-I concentrations in bull calves fed high-levels of nutrition during early life increases Sertoli cell number, supports testicular development and results in earlier puberty.

RD15537Circulating anti-Müllerian hormone concentrations in relation to age and season in male and female beluga (Delphinapterus leucas)

Gisele A. Montano, Todd R. Robeck, Karen J. Steinman and Justine K. O'Brien
pp. 1642-1652

Anti-Müllerian hormone (AMH) has been proposed as a biomarker of ovarian reserve in women and has been detected in many species; however, this is the first characterisation of AMH in a cetacean. AMH was described in males and females from the year of birth to over 30 years of age during different reproductive seasons, and a decrease in AMH in older animals (>30 years old) was detected. AMH may be of value for reproductive health monitoring of the species and as a biomarker of reproductive senescence in both sexes.

Online Early

The peer-reviewed and edited version of record published online before inclusion in an issue

Published online 21 July 2017

RD17067Genome-wide DNA methylation profile of prepubertal porcine testis

Xi Chen, Liu-Hong Shen, Li-Xuan Gui, Fang Yang, Jie Li, Sui-Zhong Cao, Zhi-Cai Zuo, Xiao-Ping Ma, Jun-Liang Deng, Zhi-Hua Ren, Zhong-Xu Chen and Shu-Min Yu

The testis is the organ essential for spermatogenesis and male fertility. This paper profiles differences in DNA methylation levels during prepubertal porcine testis development, and provides new insights into mammalian testis development and represents a foundation for additional research using the pig as a model animal.

Published online 20 July 2017

RD17041Quantitative changes of extravillous trophoblast cells in heavy smoker mothers compared with healthy controls

Zahra Heidari, Hamidreza Mahmoudzadeh-Sagheb and Nadia Sheibak

Extravillous trophoblast cells are one of the main elements that are affected by maternal smoking during pregnancy. Quantitative changes in extravillous trophoblast cells could be evidence for the hypothesis that changes in these cells could affect fetal health by changes in normal placentation.

Published online 17 July 2017

RD16338Calcium-sensing receptor (CASR) is involved in porcine in vitro fertilisation and early embryo development

C. Liu, Y. Liu, K. Larsen, Y. P. Hou and H. Callesen

Fertilisation and development of the mammalian egg is still not well understood, illustrated by low efficiencies of assisted reproductive technologies. The role of a cell membrane receptor for calcium (CASR) was studied, using pig spermatozoa and eggs as models. Significant effects were demonstrated by stimulating this receptor at different steps of the process. These findings are useful for future basic and applied research.

Inflammation plays an important role in the initiation of preterm birth, which remains the largest cause of neonatal deaths. In the present study, components of the protein synthesis machinery were significantly increased with labouring compared to non-labouring fetal membranes and myometrium, and blockade of protein synthesis significantly reduced inflammation in the myometrium. These results may provide further insight into the pathways involved in the initiation of preterm birth.

Toll-like receptors (TLRs) contribute to an exclusive protection system in the uterus during the oestrus cycle; however, the importance of TLRs in tissue repair within the involuting uterus is not known. Herein we show that TLRs are constitutively expressed in uterine cells and that the localisation pattern of TLRs in the endometrium varies with structural changes in the involuting uterus. These results suggest that TLRs have a physiological role in the uterine remodelling events during the post partum involution period.

Published online 11 July 2017

RD17045Unique oestrogen receptor ligand-binding domain sequence of native parrots: a possible link between phytoestrogens and breeding success

Catherine E. J. Davis, Adrian H. Bibby, Kevin M. Buckley, Kenneth P. McNatty and Janet L. Pitman

Hormones in plants may provide an important trigger for activating breeding activity in Kākāpō, a critically endangered New Zealand parrot. By comparing native and Australian parrots for their ability to respond to steroid hormones, we discovered novel sequences in the receptivity of parrots to oestrogenic compounds. Receptor modeling revealed that altered interactions with plant oestrogens in parrots may enhance the physiological pathways linked to fertility and successful breeding.

Published online 11 July 2017

RD17068Efficient generation of goats with defined point mutation (I397V) in GDF9 through CRISPR/Cas9

Yiyuan Niu, Xiaoe Zhao, Jiankui Zhou, Yan Li, Yu Huang, Bei Cai, Yutai Liu, Qiang Ding, Shiwei Zhou, Jin Zhao, Guangxian Zhou, Baohua Ma, Xingxu Huang, Xiaolong Wang and Yulin Chen

Taking advantages of the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system, we achieved a precise single-nucleotide substitution in the growth differentiation factor 9 (GDF9) gene using a single-stranded oligo deoxynucleotide as a homology-dependent repair template with a relatively high efficiency in order to increase litter size in goats. The present study provides an alternative approach for introducing defined point mutations in large animal models for the improvement of significant traits, as well as for the functional validation of key variants.

Published online 06 July 2017

RD16441Expanded equine cumulus–oocyte complexes exhibit higher meiotic competence and lower glucose consumption than compact cumulus–oocyte complexes

L. González-Fernández, M. J. Sánchez-Calabuig, M. G. Alves, P. F. Oliveira, S. Macedo, A. Gutiérrez-Adán, A. Rocha and B. Macías-García

Equine immature oocytes are classified as expanded or compact and are matured in the laboratory to obtain offspring; higher maturation rates are reported for expanded oocytes. The aim of this study was to assess whether differences in the maturation capacity of these oocytes is related to differences in glucose uptake, metabolism or apoptosis. Consistent metabolic and genomic differences were found between expanded and compact oocytes: compact oocytes consumed more glucose but matured less than expanded oocytes. This indicates that the maturation requirements vary for compact and expanded oocytes.

Published online 06 July 2017

RD17174Linoleic (LA) and linolenic (ALA) acid concentrations in follicular fluid of prepubertal goats and their effect on oocyte in vitro maturation and embryo development

Montserrat Roura, María G. Catalá, Sandra Soto-Heras, Sondes Hammami, Dolors Izquierdo, Ali Fouladi-Nashta and Maria-Teresa Paramio

Juvenile in vitro embryo transfer (JIVET) can increase the gain rate by reducing the generation interval, but the low quality of these oocytes leads to poor embryo production. In this study we hypothesised that oocyte quality could be related to omega 3 (linolenic acid, ALA) and omega 6 (linoleic acid, LA) concentrations in follicular fluid. No differences were found in ALA and LA concentrations in relation to follicle diameter. The addition of a high LA (omega 6) concentration to the IVM medium impaired oocyte competence to develop up to the blastocyst stage.

Published online 06 July 2017

RD17022Analysis of ZP1 gene reveals differences in zona pellucida composition in carnivores

C. Moros-Nicolás, A. Leza, P. Chevret, A. Guillén-Martínez, L. González-Brusi, F. Boué, M. Lopez-Bejar, J. Ballesta, M. Avilés and M. J. Izquierdo-Rico

The zona pellucida (ZP) is an extracellular matrix formed by few glycoproteins that covers the oocyte and plays an important role during fertilisation. The identification of ZP glycoproteins in carnivores is relevant for an understanding of sperm–ZP interactions and species specificity. In this study, molecular and in silico analyses demonstrated the presence of the ZP1 gene in four families (Felidae, Mustelidae, Odobenidae and Ursidae), whereas in three families (Canidae, Otariidae and Phocidae) ZP1 is pseudogenised. This information could be relevant for the development of more specific and efficient contraceptive vaccines for carnivores.

Published online 04 July 2017

RD16301Visfatin and resistin in gonadotroph cells: expression, regulation of LH secretion and signalling pathways

Virginie Maillard, Sébastien Elis, Alice Desmarchais, Céline Hivelin, Lionel Lardic, Didier Lomet, Svetlana Uzbekova, Philippe Monget and Joëlle Dupont

Resistin and visfatin, two adipokines, modulate reproduction in the gonads, but their actions at the hypothalamic–pituitary level are not known. We showed in vitro that these hormones decreased LH secretion from mouse gonadotroph cells and that in vivo visfatin may be a myokine as well as an adipokine in mice. Our findings suggest that these hormones may influence mouse female fertility by regulating LH secretion at the pituitary level.

Published online 23 June 2017

RD17107Delayed onset of puberty in male offspring from bisphenol A-treated dams is followed by the modulation of gene expression in the hypothalamic–pituitary–testis axis in adulthood

Isabela M. Oliveira, Renata M. Romano, Patricia de Campos, Monica D. Cavallin, Claudio A. Oliveira and Marco A. Romano

The developing fetal brain is a potent target for endocrine chemical disruption by bisphenol A (BPA) because of its lipophilicity. In the present study, perinatal exposure to BPA changed the expression of genes regulating the hypothalamic–pituitary–testis (HPT) axis at the hypothalamic, pituitary and testes levels. Maternal exposure to BPA during fetal differentiation of the sexually dimorphic nucleus of the hypothalamus caused reprogramming of the HPT axis in adult male offspring

Published online 19 June 2017

RD17170Beneficial effects of melatonin on in vitro embryo production from juvenile goat oocytes

Sandra Soto-Heras, Montserrat Roura, Maria G. Catalá, Irene Menéndez-Blanco, Dolors Izquierdo, Ali A. Fouladi-Nashta and Maria T. Paramio

Juvenile in vitro embryo transfer (JIVET) can increase the genetic gain rate by reducing the generation interval, but the low quality of the oocytes and higher oxidative levels leads to poor embryo production. In the present study we hypothesised that melatonin could be a suitable tool for improving oocyte competence in juvenile goats. The results show that melatonin is present in the follicular fluid in juvenile goats and its addition to the in vitro maturation medium reduces oxidation and increases embryo development showing its potential role for JIVET.

Published online 14 June 2017

RD16282Increased high molecular weight adiponectin, but decreased total adiponectin and kisspeptin, in central precocious puberty compared with aged-matched prepubertal girls

Chantacha Sitticharoon, Maynart Sukharomana, Supawadee Likitmaskul, Malika Churintaraphan and Pailin Maikaew

The factors initiating puberty have not been clearly revealed. The present study compared serum total and high molecular weight (HMW) adiponectin levels between girls with central precocious puberty (CPP) and age-matched controls. Total adiponectin levels were low, but HMW adiponectin levels were high in girls with CPP, which probably leads to decreased inhibition of gonadotrophin-releasing hormone neuronal activity. This may provide an insight into how adipokines are involved in the initiation of puberty.

Published online 14 June 2017

RD16290Novel insights into the role of cell-free seminal mRNAs on semen quality and cryotolerance of spermatozoa in bulls (Bos taurus)

Munivenkatappa Shilpa, Sellappan Selvaraju, Venkataswamy GirishKumar, Sivashanmugam Parthipan, Krishnan B. Binsila, Arunachalam Arangasamy and Janivara Parameshwaraiah Ravindra

The selection of fertile males is important to augment fertility in cows, and breeding soundness examinations may not provide information on the fertility potential of bulls. Expression levels of seminal plasma RNAs may be used to predict the fertilising ability and cryotolerance of bull semen. Such non-invasive tests using seminal plasma may aid in the assessment of the health of the reproductive tract and in the selection of males for breeding.

Published online 09 June 2017

RD17089Temperature gradients in vivo influence maturing male and female gametes in mammals: evidence from the cow

R. H. F. Hunter, F. López-Gatius and O. López-Albors

Is the temperature in a preovulatory follicle lower than in adjacent tissues? Using a novel non-invasive approach it was demonstrated that bovine follicular fluid before ovulation is cooler than the uterine surface and deep rectal tissues, whereas temperature is not reduced in follicles suffering ovulation failure. Follicular antral temperature could become a more valuable indicator of oocyte potential than a purely morphological assessment.

Published online 02 June 2017

RD17136Effect of seminal plasma from high- and low-fertility bulls on cauda epididymal sperm function

S. A. Holden, B. Fernandez-Fuertes, E. M. Murphy, P. Lonergan and S. Fair

Seminal plasma bathes the sperm upon ejaculation but its function has yet to be fully defined. This study sought to characterise the effect of seminal plasma from bulls of high or low fertility on sperm function. Seminal plasma reduced osmotic resistance of epididymal sperm, regardless of whether it was from high or low fertility bulls.

Published online 01 June 2017

RD16398Effects of ovarian disaggregation on adult murine follicle yield and viability

Fiona Young, John Drummond, Emma Akers, Louise Bartle, David Kennedy and Mohammad Asaduzzaman

Enzymatic disaggregation of murine ovaries yielded higher numbers of morphologically intact follicules than previously reported. The molecular markers DAPI, CMXRos and alpha-tubulin were applied to assess post-harvest follicular morphology, mitochondrial function and cellular cytoskeleton respectively. A new preparation of animal origin-free collagenase IV caused more damage to follicular integrity, mitochondrial function and the cytoskeleton than enzyme-free mechanical isolation, but another preparation of collagenase IV ovarian disaggregation gave the highest yield of morphologically intact follicles containing viable mitochondria.

Published online 30 May 2017

RD17111Splicing-related single nucleotide polymorphism of RAB, member of RAS oncogene family like 2B (RABL2B) jeopardises semen quality in Chinese Holstein bulls

Xiuge Wang, Xiaohui Cui, Yan Zhang, Haisheng Hao, Zhihua Ju, Deyu Liu, Qiang Jiang, Chunhong Yang, Yan Sun, Changfa Wang, Jinming Huang and Huabin Zhu

Bull semen quality traits are vital parameters in evaluating the fertility potential of bulls. In the present study we demonstrated that a single nucleotide polymorphism within the exonic splicing enhancer motif caused aberrant splicing of the RAB, member of RAS oncogene family like 2B (RABL2B) gene, and that semen quality differed among Chinese Holstein bulls with different RABL2B genotypes. These results also provide a novel research idea for further elucidating the complex molecular mechanism underlying the regulation of bull fertility.

Published online 23 May 2017

RD17100Mouse minipuberty coincides with gonocyte transformation into spermatogonial stem cells: a model for human minipuberty

Ruili Li, Amanda Vannitamby, Sarah S. K. Yue, David Handelsman and John Hutson

Subsequent infertility and testicular cancer are major problems for boys with undescended testis. This study aims to examine whether mouse has similar postnatal hormonal changes as boys, so it can be used as a model to study human testis development. Our results demonstrated that mouse can be used as a model for human minipuberty.

Published online 19 May 2017

RD16457Differential gene expression in porcine oviduct during the oestrous cycle

O. S. Acuña, M. Avilés, R. López-Úbeda, A. Guillén-Martínez, C. Soriano-Úbeda, A. Torrecillas, P. Coy and M. J. Izquierdo-Rico

The importance of the oviduct for different reproductive processes is known; however, the molecular mechanisms underlying oviductal regulation are not fully understood. The results of the present study demonstrate that gene expression in the oviduct is regulated during the oestrous cycle, with some oviductal proteins that could be related to several reproductive processes being described for the first time. These findings are very important in our understanding of the different processes that take place in the oviduct as fertilisation or early embryo development.

Published online 19 May 2017

RD16460Regulation of interleukin-11 expression in ovulatory follicles of the rat ovary

You-Jee Jang, You-Jee Jang, Jae-Il Park, Jae-Il Park, Seong-Eun Jeong, Seong-Eun Jeong, You-Mi Seo, You-Mi Seo, Phuong T. M. Dam, Phuong T. M. Dam, Young-Woo Seo, Young-Woo Seo, Bum-Chae Choi, Bum-Chae Choi, Sang-Jin Song, Sang-Jin Song, Sang-Young Chun, Sang-Young Chun, Moon-Kyoung Cho and Moon-Kyoung Cho

The expression of IL-11 is stimulated by LH/hCG in theca cells of preovulatory follicles via PKA-MAPK pathway and TLR4 activation. IL-11 increases progesterone production by stimulating StAR expression.

Published online 15 May 2017

RD16386Effects of vitrification of cumulus-enclosed porcine oocytes at the germinal vesicle stage on cumulus expansion, nuclear progression and cytoplasmic maturation

Ruth Appeltant, Tamás Somfai, Elisa C. S. Santos, Thanh Quang Dang-Nguyen, Takashi Nagai and Kazuhiro Kikuchi

Although oocyte cryopreservation is desired for the preservation of female germplasm, the developmental competence of vitrified porcine oocytes has been notoriously low due to unclear reasons. We investigated the effects of vitrification of porcine oocytes at the immature stage. Vitrification triggered premature nuclear resumption and nucleolar fragmentation from which some oocytes could recover, suggesting that enhancement of recovery processes might be a possible way to improve competency.

Published online 28 April 2017

RD16366Development of pre-implantation porcine blastocysts cultured within alginate hydrogel systems either supplemented with secreted phosphoprotein 1 or conjugated with Arg-Gly-Asp Peptide

Taylor D. Laughlin, Jeremy R. Miles, Elane C. Wright-Johnson, Lea A. Rempel, Clay A. Lents and Angela K. Pannier

Deficiencies in the initiation of embryo elongation play a major role in early embryonic mortality in the pig. This study demonstrates that embryos encapsulated and cultured in alginate hydrogels with the integrin-binding Arg-Gly-Asp (RGD) sequence had increased embryonic survival and modulation of gene expression to support increased steroid production and adequate immune responsiveness, with a larger number exhibiting the morphological changes compared to other in vitro treatments in this study. These results illustrate the importance of the integrin-binding RGD peptide sequence for stimulating the initiation of embryo elongation and improving embryo survival.

Published online 26 April 2017

RD16378Neuromodulatory effect of oestradiol in the metabolism of ovarian progesterone and oestradiol during dioestrus II: participation of the superior mesenteric ganglion

Adriana Vega Orozco, Cynthia Bronzi, Sandra Vallcaneras, Zulema Sosa and Marilina Casais

This work demonstrates that estradiol through the peripheral nervous system modulates the ovarian physiology, favoring the physiological luteolysis. The objective of this work was to demonstrate whether estradiol in superior mesenteric ganglion modifies the release of ovarian progesterone, estradiol, nitric oxide and noradrenaline on DII day. The ganglionic stimulation produces a decrease in the ovarian release of progesterone and estradiol by an increase of nitric oxide and decrease of noradrenaline.

Published online 26 April 2017

RD17046Protective effects of polydatin on experimental testicular torsion and detorsion injury in rats

Huilian Qiao, He Ma, Wanjun Cao, Hao Chen, Jinhua Wei and Zhen Li

The notable pathophysiological event that occurs in testicular torsion–detorsion (T/D) is ischaemia–reperfusion (I/R), which lacks efficient clinical treatment. Herein we describe, for the first time, the protective effects of polydatin, a traditional Chinese medicine, against testicular T/D injury in the rat. Our findings may provide new clues for male infertility associated with testicular torsion.

Published online 26 April 2017

RD16428Intracytoplasmic oxidative stress reverses epigenetic modifications in polycystic ovary syndrome

Fatemeh Eini, Marefat Ghaffari Novin, Khojasteh Joharchi, Ahmad Hosseini, Hamid Nazarian, Abbas Piryaei and Arash Bidadkosh

In PCOS, elevated levels of plasma androgens affects the quality of oocytes and ovulation rate, and causes anovulation in women of reproductive age. We studied the mechanism in a mouse model of PCOS, and showed that intracytoplasmic oxidative stress in oocyte via increase in histone acetylation and decrease in DNA methylation and histone dimethylation alters epigenetic modification. Consistently, mRNA expression of DNA methyltransferase-1 and histone deacetylase-1 was decreased with PCOS. There was a significant correlation between ROS production and increased histone acetylation. We concluded that antioxidant therapy in PCOS is a viable approach to reduce the adverse effects of epigenetic alterations.

Published online 19 April 2017

RD16343Andrographolide disrupts meiotic maturation by blocking cytoskeletal reorganisation and decreases the fertilisation potential of mouse oocytes

Hong-xing Liang, Sheng-sheng Lu, Zheng Yan, Yan-ping Kuang, Xiang-xing Zhu, Zhi-guang Yan, Tong Du, Wei-ran Chai, Hui Long and Qi-feng Lyu

Andrographolide should be prudently consumed by women attempting to conceive. The aim of the present study was to investigate the effects of andrographolide on oocytes. The findings indicate that andrographolide may disrupt mouse oocyte meiotic maturation and fertilisation potential by blocking cytoskeletal reorganisation. This study provides an important platform for understanding the potential adverse effects of andrographolide on female fertility.

Published online 19 April 2017

RD16459Comparative analysis of granulosa cell gene expression in association with oocyte competence in FSH-stimulated Holstein cows

David A. Landry, Chloé Fortin, Anne-Marie Bellefleur, Rémi Labrecque, François-Xavier Grand, Christian Vigneault, Patrick Blondin and Marc-André Sirard

In cattle, a major limitation of the success of IVF is the developmental competence that oocytes acquire during ovarian stimulation. The aim of the present study was to better understand the follicular conditions associated higher versus lower embryo yields. We revealed that the level of follicular differentiation (timing) is the main cause of failure and provide new biomarkers to identify such follicular conditions to adapt the ovarian stimulation protocol to individual animals.

Published online 18 April 2017

RD16435Proteomic analysis of the endometrium during early pregnancy in the domestic pig

Justyna Kolakowska, Serhiy Souchelnytskyi, Ravi Kanth Rao Saini and Anita Franczak

High embryo loss during early pregnancy in pigs is still an unresolved issue. Comparing the proteomic profile of the endometrium harvested from crucial stages of early pregnancy (Days 12–13 and 15–16) with the proteomic profile of tissue from corresponding days of the oestrous cycle revealed the presence of proteins unique for early pregnancy that are involved in adhesion, angiogenesis and cytoskeletal organization. The results of the present study may help us understand the causes of early pregnancy loss in pigs.

Binding of spermatozoa to the egg coat is a key step in fertilisation and provides a mechanism for reproductive isolation and sexual selection. To test this theory we investigated the molecular evolution of the ZP3 gene and its encoded sperm-binding region in a diverse murine rodent group. Our analyses provides evidence for positive selection on the sperm-binding region in several murine lineages, but the nature of the selective force remains unresolved.

Published online 12 April 2017

RD16498Endo-siRNA deficiency results in oocyte maturation failure and apoptosis in porcine oocytes

Wanxin Liu, Qi Zhao, Shanhua Piao, Chunsheng Wang, Qingran Kong and Tiezhu An

Endo-siRNAs play an important role in mouse oocyte meiosis but the function of endo-siRNAs in other species is unknown. In this study, we found that an endo-siRNA deficiency resulted in oocyte spindle abnormalities and apoptosis, suggesting that endo-siRNAs are indispensible for oocyte maturation in pigs. This study unequivocally demonstrates an essential function for endo-siRNAs in mammals.

Published online 10 April 2017

RD16267Pannexin channels increase propidium iodide permeability in frozen–thawed dog spermatozoa

J. L. Torres, J. Palomino, R. D. Moreno and M. De los Reyes

Pannexins (Panx) are proteins that form functional single membrane channels that have not yet been described in dogs. The aim of the present study was to detect Panx1, Panx2 and Panx3 in frozen–thawed dog semen. All three Panx channels were present in dog spermatozoa and increased propidium iodide (PI) permeability in frozen–thawed dog spermatozoa, suggesting that the percentage of PI-positive spermatozoa used as an indicator of non-viable cells may lead to overestimation of non-viable cells.

Published online 10 April 2017

RD16474Differential effects of high and low glucose concentrations during lipolysis-like conditions on bovine in vitro oocyte quality, metabolism and subsequent embryo development

J. De Bie, W. F. A. Marei, V. Maillo, L. Jordaens, A. Gutierrez-Adan, P. E. J. Bols and J. L. M. R. Leroy

Maternal metabolic disorders (e.g. obesity and diabetes in women or a negative energy balance in dairy cows) are associated with altered metabolic profiles, which can impact oocyte quality and thus fertility. We showed that altered glucose can differentially affect oocyte quality in the presence of elevated free fatty acids. These results further highlight the importance of maternal metabolic health for fertility.

Published online 10 April 2017

RD16442Supplementation of culture medium with L-carnitine improves the development and cryotolerance of in vitro-produced porcine embryos

J. L. Lowe, L. K. Bartolac, R. Bathgate and C. G. Grupen

Porcine embryos contain substantial amounts of lipid, with little known regarding its metabolic role during development. This study investigated the association of lipid metabolism with carbohydrate substrates in the culture medium and revealed positive effects of adding L-carnitine, a limiting co-factor required for lipolysis, on early cleavage and blastocyst quality. Supplementing medium with L-carnitine is a useful strategy to improve the success of porcine blastocyst vitrification.

Published online 07 April 2017

RD16432Low doses of bisphenol A can impair postnatal testicular development directly, without affecting hormonal or oxidative stress levels

Fernanda M. Ogo, Glaucia E. M. L. Siervo, Géssica D. Gonçalves, Rubens Cecchini, Flavia A. Guarnier, Janete Ap. Anselmo-Franci and Glaura S. A. Fernandes

The peripubertal period is considered a critical stage of reproductive development and more vulnerable to the action of toxic agents, such as BPA, which can lead to impaired sexual development. We investigated the exposure to BPA during the peripubertal period and eEvidencing a reduction in the number of Sertoli cells, histopathological abnormalities and changes in sperm morphology. Therefore, BPA may silently impair testicular postnatal development in rats.

Published online 06 April 2017

RD16265Sex-specific differences and developmental programming for diseases in later life

Deepali P. Sundrani, Suchitra S. Roy, Anjali T. Jadhav and Sadhana R. Joshi

Sex-specific differences have been reported in most of the diseases. This review provides data on differential adaptation of male and female fetuses in utero in response to maternal nutritional, physiological and environmental insults resulting in sex-specific differences. Because sex differences are associated with fetal outcome and survival, it is of importance to study the underlying biological mechanisms to improve both pre-natal and neonatal care.

Published online 05 April 2017

RD16235Endogenous lysophosphatidic acid participates in vascularisation and decidualisation at the maternal–fetal interface in the rat

Micaela S. Sordelli, Jimena S. Beltrame, Elsa Zotta, Natalia Gomez, Ganna Dmytrenko, María Elena Sales, Sandra M. Blois, Carlos Davio, Silvina Perez Martinez, Ana M. Franchi and María L. Ribeiro

Decidualisation and vascular remodelling are fundamental for placentation and pregnancy success. Disruption of endogenous lysophosphatidic acid signalling modified the development of uterine vessels with consequences in the formation of the decidua and placenta, compromising the growth of embryos. Defects in these mechanisms could appear as obstetric complications, like implantation failure, positioning lysophosphatidic acid as a regulatory key lipid at implantation.

Published online 03 April 2017

RD16513Melatonin potentially acts directly on swine ovary by modulating granulosa cell function and angiogenesis

Giuseppina Basini, Simona Bussolati, Roberta Ciccimarra and Francesca Grasselli

The reproductive effects of melatonin have been extensively studied, but its involvement in the local control of ovarian physiology has not yet been clarified. The present research, which was undertaken to study the effects of physiological levels of melatonin on the main parameters of swine granulosa cell function, documents that the hormone is involved in the modulation of ovarian follicle growth and development.

Successful artificial fertilisation primarily depends on two characteristics of male quality: 1) the maintenance of high sperm viability during storage and 2) the effective activation of sperm motility. This study identified optimal conditions for sperm motility activation in the critically endangered booroolong frog. Results reported make a significant contribution to the growing field of amphibian reproduction technologies.

Published online 30 March 2017

RD16320Steroid hormones interact with natriuretic peptide C to delay nuclear maturation, to maintain oocyte–cumulus communication and to improve the quality of in vitro-produced embryos in cattle

Ana Caroline S. Soares, Valentina Lodde, Rodrigo G. Barros, Christopher A. Price, Alberto M. Luciano and José Buratini

This study provides evidence that natriuretic peptide C and steroids interact to slow oocyte nuclear maturation and to enhance oocyte-cumulus communication in cattle. Moreover, it indicates that exposure of the cumulus–oocyte complex to physiological concentrations of these hormones is beneficial to oocyte developmental competence. Therefore, the dataset is of relevance for the progress of oocyte biology and for the improvement of in vitro maturation outcomes.

Published online 30 March 2017

RD16511Cryopreservation of saltwater crocodile (Crocodylus porosus) spermatozoa

S. D. Johnston, E. Qualischefski, J. Cooper, R. McLeod, J. Lever, B. Nixon, A. L. Anderson, R. Hobbs, J. Gosálvez, C. López-Fernández and T. Keeley

A combination of permeating and non-permeating cryoprotectants and freezing rates was used to successfully cryopreserve saltwater crocodile spermatozoa. Although glycerol concentrations >1.35 M had an inhibitory effect on motility before freezing, 0.68 M glycerol combined with 0.2 M sucrose and post-thaw washing of spermatozoa in capacitating media resulted in motility of between 12% and 14.2%, the highest ever recorded for this species.

Published online 30 March 2017

RD16475Placentation in the plains zebra (Equus quagga)

W. R. (Twink) Allen, Fiona Stansfield and Sandra Wilsher

This paper describes findings during a unique opportunity to monitor placental structure and function throughout pregnancy in the under-researched zebra. Zebras are more donkey-like than horse-like in placental structure, rate of fetal development and gestation length. The results highlight the retention of unique reproductive mechanisms across the Genus Equus despite many other evolutionary changes within the member species.

Published online 27 March 2017

RD16289Successful chimera production in the Hungarian goose (Anser anser domestica) by intracardiac injection of blastodermal cells in 3-day-old embryos

Nikoletta Sztán, Bence Lázár, Nóra Bodzsár, Barbara Végi, Krisztina Liptói, Bertrand Pain and Eszter Patakiné Várkonyi

The aim of this study was to develop a new genome preservation technique for goose species. According to our results, goose primordial germ cells (PGCs) migrate between 69 and 84 h of development. The goose blastoderm contains PGCs and by isolating and injecting these blastodermal cells we were able to generate chimeras. Altogether, this method could contribute to the cryopreservation of this species.

Published online 27 March 2017

RD17020Glutamine protects rabbit spermatozoa against oxidative stress via glutathione synthesis during cryopreservation

Zhendong Zhu, Xiaoteng Fan, Yinghua Lv, Yan Lin, De Wu and Wenxian Zeng

The addition of 20 mM glutamine to the freezing extender significantly improved post-thaw sperm motility, acrosome integrity, membrane integrity and mitochondrial activity. The exposure of spermatozoa to glutamine enhanced their γ-GCS activity and GSH synthesis, while decreasing ROS accumulation, LPO and DNA damage. Therefore, glutamine protects spermatozoa against ROS-induced cryodamage by enhancing GSH synthesis.

Published online 16 March 2017

RD16144Prostaglandin F–PTGFR signalling activation, growth factor expression and cell proliferation in bovine endometrial explants

Shuangyi Zhang, Bo Liu, Long Gao, Wei Mao, Changqi Fu, Duritahala, Nan Zhang, Ying Zhang, Yuan Shen and Jinshan Cao

The association between bovine endometrial growth and prostaglandin (PG) F–PGF receptor (PTGFR) signalling activation is unknown. In the present study, cell proliferation and the expression of growth factors essential for endometrial growth were upregulated by PGF-PTGFR signalling activation by fluprostenol in bovine endometrial explants in vitro. These results indicate that PGF-PTGFR signalling activation could be involved in endometrial growth.

uPA shRNA lentivirus system induced by Dox inhibited the expression of uPA in TM4 cells. Through injecting the lentivirus into mice testis, we found that downregulation of uPA in mice testis decreased the fertility of male mice, which may be caused by a reduction in sperm motility. The inducible uPA shRNA system is safe and reversible to regulate male fertility.

Live cell RNA imaging in mammalian embryos would represent a quantum leap in understanding cell differentiation and early embryonic development. Applicability of RNA imaging by SmartFlare technology was tested in equine embryos surrounded by an embryonic capsule and/or the zona pellucida, nude trophoblastic vesicles and fibroblast cell cultures. Encapsulated embryos block SmartFlare uptake, but stages surrounded by the zona pellucida only and nude vesicles incorporate the probes, leading to specific fluorescence suitable for tracing gene expression cascades necessary for further embryo development.

Published online 28 February 2017

RD16426Effects of antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification on the in vitro developmental capacity of expanded bovine blastocysts

Shuang Liang, Bao Yuan, Yong-Xun Jin, Jia-Bao Zhang, Jeong Kyu Bang and Nam-Hyung Kim

Antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification beneficially protects bovine blastocysts against chill-induced injury. AFGP8 supplementation during vitrification enhances the cryosurvival of bovine blastocysts. AFGP8 supplementation during vitrification improves the subsequent in vitro developmental capacity of bovine blastocysts.

Published online 28 February 2017

RD16478Expression of protocadherin 11Yb (PCDH11Yb) in seminal germ cells is correlated with fertility status in men

Thottathil R. Anilkumar, Anandavalli N. Devi, Sathy M. Pillai, Krishnapillai Jayakrishnan, Oommen V. Oommen and Pradeep G. Kumar

Knowledge of molecular defects in spermatozoa becomes critical during assisted reproduction to treat male factor infertility. Herein we show that the absence or low expression of protocadherin 11Yb (PCDH11Yb) in spermatozoa is associated with male factor infertility. Profiling of a panel of differentially expressed sperm proteins, including PCDH11Yb, in men with infertility may be of diagnostic and therapeutic value in infertility management.

Published online 24 February 2017

RD16383Localisation of stem cell factor, stanniocalcin-1, connective tissue growth factor and heparin-binding epidermal growth factor in the bovine uterus at the time of blastocyst formation

M. Muñoz, D. Martin, S. Carrocera, M. Alonso-Guervos, M. I. Mora, F. J. Corrales, N. Peynot, C. Giraud-Delville, V. Duranthon, O. Sandra and E. Gómez

Limited information is available about the role of growth factors in very early embryonic development. This study demonstrates the presence of stem cell factor, stanniocalcin-1, connective tissue growth factor and heparin-binding epidermal growth factor-like growth factor in bovine uterine fluid, endometrium and embryos during blastocyst formation. The results suggest a role for these proteins in early embryo development in cattle.

Published online 14 February 2017

RD16186Identification of differentially expressed placental transcripts during multiple gestations in the Eurasian beaver (Castor fiber L.)

A. Lipka, L. Paukszto, M. Majewska, J. P. Jastrzebski, K. Myszczynski, G. Panasiewicz and B. Szafranska

It is still unspecified if and how the number of embryos or fetal sex affect pregnancy outcome. The present study determined whether there are differences in transcriptome profiles of the beaver placenta as a result of fetal sex or multiplicity of gestation. The results indicate that the number of fetuses affects the expression profile in the subplacental transcriptome. Enhancement of transcriptome resources will improve our understanding of the pathways relevant to proper placental development and successful reproduction in general.

Published online 14 February 2017

RD16302Polydatin improves the developmental competence of bovine embryos in vitro via induction of sirtuin 1 (Sirt1)

Imran Khan, Sung Woo Kim, Kyung-Lim Lee, Seok-Hwan Song, Ayman Mesalam, M. M. R. Chowdhury, Zia Uddin, Ki Hun Park and Il-Keun Kong

Polydatin (PD) has antioxidant activity and is useful for embryo development in vitro. Supplementation of IVM medium with PD scavenged reactive oxygen species and improved embryo developmental competence via induction of sirtuin 1 (Sirt1).

Published online 13 February 2017

RD16294Sperm gamma-aminobutyric acid type A receptor delta subunit (GABRD) and its interaction with purinergic P2X2 receptors in progesterone-induced acrosome reaction and male fertility

Wenming Xu, Ke Wang, Yan Chen, Xiao Tong Liang, Mei Kuen Yu, Huanxun Yue and M. Louise Tierney

The mechanism underlying the non-genomic action of progesterone in sperm functions and related Ca2+ mobilisation remains elusive. Herein we report the expression of gamma-aminobutyric acid type A receptor delta subunit (GABRD) in human and rodent spermatozoa and its involvement in mediating the progesterone-induced acrosome reaction through its interaction with purinergic P2X2 receptors. GABRD represents a novel progesterone receptor or modulator in sperm responsible for the progesterone-induced Ca2+ influx required for the acrosome reaction.

Published online 10 February 2017

RD16333Restoration of immune and renal function in aged females by re-establishment of active ovarian function

Rhett L. Peterson, Kate C. Parkinson and Jeffrey B. Mason

Immune and renal function, both critical for reproductive success, were evaluated to determine if exposure of postreproductive female mice to young ovaries would influence these non-reproductive physiological functions. Postreproductive mice received new (60-day-old) ovaries at 12 months of age and were evaluated at 16 months of age. Age-related declines in immune and renal function in 16-month-old control mice were restored to levels found in 6-month-old mice by ovarian transplantation. Re-establishment of reproductive function in aged female mice positively influenced non-reproductive functions.

Published online 08 February 2017

RD16473In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen–thawed stallion semen

D. M. Silva, S. A. Holden, A. Lyons, J. C. Souza and S. Fair

Improvements in the quality of stallion semen are required to increase the success of artificial insemination. The aim of this study was to assess the effects of the addition of docosahexaenoic acid to semen diluents on the quality of stallion semen. Docosahexaenoic acid increased spermatozoa motility and membrane fluidity in cooled semen and thus can be used to increase the quality of spermatozoa.

Published online 03 February 2017

RD16327Gene-specific profiling of DNA methylation and mRNA expression in bovine oocytes derived from follicles of different size categories

F. Mattern, J. Heinzmann, D. Herrmann, A. Lucas-Hahn, T. Haaf and H. Niemann

The present study investigated DNA methylation profiles and mRNA expression of imprinted and non-imprinted candidate genes in bovine oocytes from antral follicles of three different size classes (≤2 mm, 3–5 mm, ≥6 mm) to unravel the epigenetic contribution to follicular and oocyte growth. We observed an increased number of aberrantly methylated alleles in bH19, bSNRPN and bDNMT3 Lo of oocytes from small antral follicles and an increased frequency of CpG sites with an unclear methylation status for DNMT3 Ls in large follicles. Results indicate that a follicle diameter of ~2 mm is critical for establishing DNA methylation profiles.

Published online 02 February 2017

RD16193Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification

Jakkhaphan Pitchayapipatkul, Tamás Somfai, Satoko Matoba, Rangsan Parnpai, Takashi Nagai, Masaya Geshi and Thevin Vongpralub

The aim of this study was to compare the efficacy of paclitaxel (PT) and docetaxel (DT) in reducing spindle damage in bovine oocytes during vitrification. The important finding of the study was that pretreatment of bovine oocytes with 0.05 µM DT for 30 min before vitrification reduced spindle damage to a greater degree than pretreatment with 1.0 µM PT. This finding will be useful in improving oocyte cryopreservation in the future.

Published online 01 February 2017

RD16233Extracellular-like matrices and leukaemia inhibitory factor for in vitro culture of human primordial follicles

Assiel J. Younis, Galit Lerer-Serfaty, Dana Stav, Bethsabee Sabbah, Tzippy Shochat, Gania Kessler-Icekson, Muayad A. Zahalka, Michal Shachar-Goldenberg, Avi Ben-Haroush, Benjamin Fisch and Ronit Abir

The possibility of growing immature human ovarian follicles in laboratories will assist fertility restoration. The aim of the present study was to investigate whether growth of immature follicles can be promoted by using different culture surfaces and supplementation with a growth factor. It was unclear whether any of the culture surfaces had an advantage over the others. In addition, the growth factor did not promote growth. Further studies should test other culture surfaces and identify additional fluid supplements, so that an optimal system can be developed.

Poly(A)-binding proteins function in the timely regulation of gene expression during oocyte maturation, fertilisation and early embryo development in vertebrates. In this review, we comprehensively evaluate and discuss the expression patterns and particular functions of the embryonic poly(A)-binding protein (EPAB) and poly(A)-binding protein, cytoplasmic 1 (PABPC1) genes, especially in mouse and human oocytes and early embryos.

Published online 12 January 2017

RD16276Epigenetic changes of hepatic glucocorticoid receptor in sheep male offspring undernourished in utero

Stella Chadio, Basiliki Kotsampasi, Stylliani Taka, Emmanouil Liandris, Nikolaos Papadopoulos and Elias Plakokefalos

Evidence exists that early life nutritional insults can predispose to metabolic disorders, like Type 2 diabetes, for which excessive hepatic glucose production is a contributing factor. Maternal undernutrition alters the epigenetic status of the glucocorticoid receptor and increases the expression of gluconeogenic enzymes in the liver of male offspring. Undernutrition in the womb programs long-lasting changes in specific gene expression that leads to persistent metabolic abnormalities.

Published online 09 January 2017

RD16277Maternal obesity in the rat impairs male offspring aging of the testicular antioxidant defence system

Claudia J. Bautista, Guadalupe L. Rodríguez-González, Angélica Morales, Consuelo Lomas-Soria, Fabiola Cruz-Pérez, Luis A. Reyes-Castro and Elena Zambrano

Maternal obesity leads to premature aging of male offspring reproductive capacity and increases testes oxidative stress in male rat offspring. Our results show that maternal obesity modifies the antioxidant enzyme system in the testes of offspring, which, in turn, affects testicular physiological functions and leads to premature aging of male reproductive capacity.

Published online 09 January 2017

RD16348Mitochondrial ferritin deficiency reduces male fertility in mice

Federica Maccarinelli, Maria Regoni, Fernando Carmona, Maura Poli, Esther G. Meyron-Holtz and Paolo Arosio

Mitochondrial ferritin is an iron-storage protein highly expressed in human and mouse spermatozoa. This work showed that male mice, but not female, in which mitochondrial ferritin was genetically deleted have reduced fertility without significant alterations in sperm parameters. These data show that mitochondrial ferritin has a role in male fertility.

Published online 21 December 2016

RD16198Ontogeny of clock and KiSS-1 metastasis-suppressor (Kiss1) gene expression in the prepubertal mouse hypothalamus

Cassandra C. Yap, Peter J. Mark, Brendan J. Waddell and Jeremy T. Smith

Kisspeptin neurons in the brain are crucial for the hormone surge required for ovulation, but are established before puberty. This study examined kisspeptin and brain clock gene rhythms (known to restrict kisspeptin action) in young mice. The data show that the kisspeptin peak is not seen prior to puberty and there is incomplete development of clock gene rhythmicity at this time.

Published online 21 December 2016

RD16284Intrafollicular oestradiol production, expression of the LH receptor (LHR) gene and its isoforms, and early follicular deviation in Bos indicus

S. Wohlres-Viana, E. K. N. Arashiro, M. A. Machado, L. S. A. Camargo, L. G. B. Siqueira, M. P. Palhao and J. H. M. Viana

In this study we investigated the expression of the LH receptor (LHR) gene during dominant follicle selection in Bos indicus cattle. The main finding was that the early deviation of the dominant follicle in this subspecies is driven by the differential expression of full-length LHR and its isoforms in small follicles. This finding will contribute to explaining the differences in ovarian physiology between B. indicus and Bos taurus.

Published online 16 December 2016

RD16087Gel-coated tubes extend above-freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilised offspring

Brandon K. Hopkins, Susan W. Cobey, Charles Herr and Walter S. Sheppard

Honey bees, which are vital agricultural pollinators, are at risk from numerous factors. The ability to artificially inseminate these species provides a means to strengthen them genetically by collecting semen from bees of different regions throughout the world and then inseminating bees at distant locations. Our study aimed to substantially improve the viability of spermatozoa between collection and insemination.

Preferential transportation of essential fatty acids over non-essential fatty acids occurs from the mother to fetus in humans. However, little is known about placental transfer of fatty acids in the bovine. In the present study, prepartum dietary fat supplementation of cows reduced maternal long-chain polyunsaturated fatty acids, neonatal total omega-3 fatty acids, eicosapentaenoic and docosahexaenoic acids and total, fat possibly through reduced expression of certain placental fatty acid transporter genes.

Published online 12 December 2016

RD16321Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles

Tra M. T. Bui, Tra M. T. Bui, Khánh X. Nguy?n, Khánh X. Nguy?n, Asako Karata, Asako Karata, Pilar Ferré, Pilar Ferré, Minh T. Tr?n, Minh T. Tr?n, Takuya Wakai, Takuya Wakai, Hiroaki Funahashi and Hiroaki Funahashi

Ovaries contain a large number of small follicles, but the meiotic and developmental competence of oocytes from these follicles is quite low. In the present study we demonstrated that the addition of 200 ng mL–1 vascular endothelial growth factor to the IVM medium increased the maturation rate of porcine oocytes from small follicles and that blastocyst formation following parthenogenetic activation also increased. These findings may contribute to efficient animal production and human assisted reproductive technology.

Published online 12 December 2016

RD16181Modulation of adiponectin system expression in the porcine uterus during early pregnancy by prostaglandin E2 and F2?

Kamil Dobrzyn, Nina Smolinska, Karol Szeszko, Marta Kiezun, Anna Maleszka and Tadeusz Kaminski

Adiponectin is a hormone perceived as the link between energy metabolism and the reproductive system, but its interactions with factors responsible for regulation of uterine homeostasis during pregnancy remain unknown. Herein we demonstrated the effects of prostaglandins E2 and F on adiponectin and its receptors during early gestation. Recognition of these interactions will contribute to our understanding of the physiology of early pregnancy.

Published online 06 December 2016

RD15542Cholesterol metabolism in rabbit blastocysts under maternal diabetes

S. Mareike Pendzialek, Maria Schindler, Torsten Plösch, Jacqueline Gürke, Elisa Haucke, Stefanie Hecht, Bernd Fischer and Anne Navarrete Santos

A diabetic pregnancy leads to distinct changes in maternal cholesterol metabolism, whereas embryonic cholesterol concentrations and cholesterol metabolism genes are not affected. The trophoblast : embryoblast cholesterol ratio suggests a higher metabolic activity of the embryoblast in diabetes. The findings of the present study imply independent and functional cholesterol metabolism in the mammalian embryo before implantation.

Published online 06 December 2016

RD16382The current state of reproductive biology research in Australia and New Zealand: core themes from the Society for Reproductive Biology Annual Meeting, 2016

L. K. Akison, P. H. Andraweera, M. J. Bertoldo, H. M. Brown, J. S. M. Cuffe, T. Fullston, O. Holland and J. E. Schjenken

Reproductive science is central to our understanding of biology, translating to biological processes across many organ systems. The Society for Reproductive Biology conference review provides a focused summary of symposia at the 2016 annual meeting. The presentations highlight the breadth of reproductive research in Australia and New Zealand and its potential to affect the fertility and health of future generations.

Published online 28 November 2016

RD16213Protein in culture and endogenous lipid interact with embryonic stages in vitro to alter calf birthweight after embryo vitrification and warming

E. Gómez, S. Carrocera, S. Uzbekova, D. Martín, A. Murillo, M. Alonso-Guervós, F. Goyache and M. Muñoz

Intracellular lipid is involved in long-term blastocyst competence to survive vitrification. We studied how protein removal affects bovine embryo development in vitro, lipid content and pregnancy outcomes, and found that calf weight at birth was altered depending on specific embryonic kinetics within vitrified–warmed embryos. Therefore, it is possible to modify calf phenotypes by introducing simple changes in culture conditions combined with selection of embryos.

Published online 23 November 2016

RD16174Rosmarinic acid reverses the effects of metronidazole-induced infertility in male albino rats

Zina M. Al-Alami, Ziad A. Shraideh and Mutasem O. Taha

Metronidazole, an essential medicine, adversely affects male fertility. We aimed at investigating the capability of Rosmarinic acid to reverse metranidazole induced male infertility. We found that certain doses of Rosmarinic acid could reverse metronidazole’s effects on sperm count, motility, morphology and testis ultrastructure. Our findings have great clinical implications where Rosmarinic acid might have a potential use in reversing Metronidazole induced male infertility.

Published online 17 November 2016

RD15504Regulation of heat-inducible HSPA1A gene expression during maternal-to-embryo transition and in response to heat in in vitro-produced bovine embryos

Jean-Marc Lelièvre, Nathalie Peynot, Sylvie Ruffini, Ludivine Laffont, Daniel Le Bourhis, Pierre-Marie Girard and Véronique Duranthon

Early mammalian embryos are sensitive to heat stress (HS) despite abundant oocytic iHSPA protein chaperones and mRNAs typically induced by stress. In this work, bovine HSPA1A gene transcription was detectable during embryo genome activation but induced by HS only after this stage, which coincides with better embryonic tolerance. Absence of stress-induced gene expression may therefore contribute to embryo sensitivity.

Published online 11 November 2016

RD16297Effect of nutritionally induced hyperlipidaemia on in vitro bovine embryo quality depends on the type of major fatty acid in the diet

Waleed F. A. Marei, Maria Arias Alvarez, Veerle Van Hoeck, Alfonso Gutierrez-Adan, Peter E. J. Bols and Jo L. M. R. Leroy

Hyperlipidemia associated with unbalanced diet and obesity is directly linked with reduced fertility. This study shows that substituting saturated fat with omega-3 fat in high caloric diets can alleviate detrimental effects of hyperlipidemic serum on bovine early embryo development and quality in vitro. This suggests that dietary manipulation of serum fatty acid profile may enhance fertility in patients with metabolic disorders.

Published online 02 November 2016

RD16154Gene expression profile in heat-shocked Holstein and Nelore oocytes and cumulus cells

J. S. Ticianelli, I. P. Emanuelli, R. A. Satrapa, A. C. S. Castilho, B. Loureiro, M. J. Sudano, P. K. Fontes, R. F. P. Pinto, E. M. Razza, R. S. Surjus, R. Sartori, M. E. O. A. Assumpção, J. A. Visintin, C. M. Barros and F. F. Paula-Lopes

The deleterious effects of heat stress on fertility are more pronounced in dairy cows than beef cows. The transcriptome profile in Nelore and Holstein oocytes subjected to heat shock during IVM and the mRNA abundance of selected genes in cumulus cells were determined. The analyses indicated that the deficiency in dairy cattle in terms of cellular protection against heat stress is regulated at the post-transcriptional level.

Published online 26 October 2016

RD15535Sleep restriction in Wistar rats impairs epididymal postnatal development and sperm motility in association with oxidative stress

Gláucia E. M. L. Siervo, Fernanda M. Ogo, Aline D. Valério, Thamara N. X. Silva, Larissa Staurengo-Ferrari, Tathiana A. Alvarenga, Rubens Cecchini, Waldiceu A. Verri, Flávia A. Guarnier, Monica L. Andersen and Glaura S. A. Fernandes

Sleep alterations are a public health problem worldwide that affect adolescents during puberty because of early school hours and interactive activities. In the present study, sleep restriction during peripuberty in rats affected epididymal postnatal development and sperm motility, in association with oxidative stress and a decrease in the epithelial compartment in the cauda epididymidis. The results of this study suggest a possibility for improving fertility after sleep restriction.

Published online 26 October 2016

RD16223Effect and possible mechanisms of melatonin treatment on the quality and developmental potential of aged bovine oocytes

Shuang Liang, Jing Guo, Jeong-Woo Choi, Nam-Hyung Kim and Xiang-Shun Cui

This research expands our understanding of the regulators that delay oocyte aging. Treatment of aged oocytes with exogenous melatonin improves oocyte quality, rearranges the cytoskeleton and improves developmental capacity. Melatonin is a good agent for delaying oocyte aging in vitro and preventing poor developmental competence of bovine oocytes in assisted reproductive technologies.

Published online 19 October 2016

RD16183Induced sub-lethal oxidative damage affects osmotic tolerance and cryosurvival of spermatozoa

Franziska Ertmer, Harriëtte Oldenhof, Saskia Schütze, Karl Rohn, Willem F. Wolkers and Harald Sieme

If the physiological balance between production and scavenging of reactive oxygen species (ROS) is shifted towards production of ROS this may result in accumulation of cell damage. The aim of this study was to establish whether sub-lethal oxidative damage negatively affects cryosurvival and pre-freeze sperm osmotic tolerance. Generation of defined levels of intracellular ROS can be used for testing strategies designed to counteract oxidative damage.

Published online 19 October 2016

RD16310Reproductive dysfunction after mercury exposure at low levels: evidence for a role of glutathione peroxidase (GPx) 1 and GPx4 in male rats

Caroline S. Martinez, Franck M. Peçanha, Daniela S. Brum, Francielli W. Santos, Jeferson L. Franco, Ana Paula P. Zemolin, Janete A. Anselmo-Franci, Fernando B. Junior, María J. Alonso, Mercedes Salaices, Dalton V. Vassallo, Fábio G. Leivas and Giulia A. Wiggers

Mercury has been identified as a risk factor for declines in sperm quality and infertility in humans. The present study delves further into the mechanisms of action of mercury, showing a role for glutathione peroxidase (GPx) 1 and GPx4 in the reproductive toxicity of mercury. The involvement of the glutathione system highlights this antioxidant system as a potential target of mercury after its accumulation in reproductive organs.

Published online 14 October 2016

RD16037Ovariectomy increases the phenotypic plasticity of the female prostate epithelium in the Mongolian gerbil (Meriones unguiculatus)

Marianna Zanatelli, Ana M. G. Custodio, Manoel F. Biancardi, Fernanda C. A. Santos, Rejane M. Góes, Luiz R. Falleiros-Jr, Patricia S. L. Vilamaior and Sebastião R. Taboga

After hormonal deprivation the female gerbil prostate undergoes several morphophysiological changes. This work identified and characterised the cellular heterogeneity of the female prostate epithelium in normal conditions and after ovariectomy. Secretory cells of the female prostate are not morphologically and functionally uniform, presenting a phenotypical plasticity according to the hormonal environment in which they operate.

The current literature does not provide a comprehensive profile of the factors involved in placental development in rats during intrauterine trophoblast migration. In this study we evaluated the gene and/or protein expression of several placental factors during the second half of pregnancy in rats and demonstrated that the expression of proteases and immunological, angiogenic, hormonal and apoptotic mediators is variable and depends on the stage of trophoblast migration. These changes in the expression profile are probably related to alterations in placental morphogenesis and function.

Published online 11 October 2016

RD16232Influence of hyaluronan on endometrial receptivity and embryo attachment in sheep

Waleed F. A. Marei, D. Claire Wathes, Kabir A. Raheem, Omnia Mohey-Elsaeed, Fataneh Ghafari and Ali A. Fouladi-Nashta

Infusion of the ovine uterus with hyaluronan (HA) during the pre-attachment period changes the expression of some adhesion markers at the embryo–maternal interface, resulting in reduced endometrial receptivity and failure of embryo attachment. In contrast, local inhibition of HA production in the uterus enhances embryo attachment.

Published online 05 October 2016

RD16200Non-surgical sterilisation methods may offer a sustainable solution to feral horse (Equus caballus) overpopulation

Sally Elizabeth Hall, Brett Nixon and R. John Aitken

Culling feral horses is currently met with public criticism, which has created an urgent need to develop new methods of population control. Delivery of a contraceptive agent that causes sterility, without the need for surgery, is currently under investigation. These novel techniques may provide a sustainable and effective alternative for feral horse control.

Published online 28 September 2016

RD15553In vivo and in vitro maturation of rabbit oocytes differently affects the gene expression profile, mitochondrial distribution, apoptosis and early embryo development

M. Arias-Álvarez, R. M. García-García, J. López-Tello, P. G. Rebollar, A. Gutiérrez-Adán and P. L. Lorenzo

Understanding the cellular and molecular mechanisms implicated in oocyte competence during rabbit in vivo and in vitro maturation is necessary to improve IVM systems in these species. This study showed for the first time that gene expression profile of in vivo-matured rabbit COCs is different from that of IVM COCs and these changes are clearly accompanied by stage-specific cytoplasmic remodelling of mitochondria, a lower rate of apoptosis of in vivo-matured CCs and better early embryo development, indicating that oocyte quality is compromised during IVM.

Published online 28 September 2016

RD16063Proliferation 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 and Qiang Weng

Male seasonal breeders show synchronised cycles of testicular growth and involution throughout the breeding cycle. The aim of the present study was to elucidate the regulatory role of cell proliferation and apoptosis in this dynamic testicular development. The study demonstrates, for the first time, that both cell proliferation and apoptosis are stimulated during the prehibernation period in wild Daurian ground squirrels. These findings advance our current understanding of the regulatory mechanisms of seasonal reproduction.

Diabetes has a profound impact on women’s reproductive health. We aimed to investigate the protective effect of fish oil and wheat-germ oil supplementation on ovarian function in streptozotocin-diabetic rats. These natural supplements counteracted oxidative stress and inflammation induced by diabetes and regained ovarian function by increasing FSH, E2, LH and AMH levels and restoring follicle counts.

Published online 28 September 2016

RD16164Testicular parameters and spermatogenesis in different birthweight boars

P. A. Auler, G. H. F. A. Moreira, C. O. Hogg, C. J. Ashworth, F. P. Bortolozzo, H. Chiarini-Garcia and F. R. C. L. Almeida

Reports of the effects of birthweight on testicular development and implication for sperm production in boars are scarce. These effects were investigated in this study. It seems that high birthweight boars have the potential to produce more spermatozoa and semen doses per ejaculate. Hence, the selection of those boars would be predictive of better lifetime productivity in the boar stud.

The antioxidant defences of epididymal fluid and spermatozoa are modulated from the caput to cauda epididymidis in a region-specific manner in the goat (Capra hircus), and loss of some of the antioxidant enzymes from spermatozoa during their epididymal transit is compensated for through a simultaneous increase in the expression and/or activity of these enzymes in the epididymal fluid of the respective segments.

Published online 21 September 2016

RD15486Discs large homologue 1 (Dlg1) coordinates mouse oocyte polarisation during maturation

Jun-Chao Wang, Hong Lv, Ke-Liang Wu, Yun-Shan Zhang, Hai-Ning Luo and Zi-Jiang Chen

Mouse oocyte maturation is a unique asymmetric division and a lot of polarity proteins may play vital roles during this progress. Our study described an evolutionarily conserved polarity protein Discs-large homologue 1 becomes highly polarized by associating with cytoskeleton and lose-of-function of Dlg1 compromises oocyte maturation. It may provide a new theoretic framework for mammalian meiosis research.

Published online 21 September 2016

RD16238Imprinted and DNA methyltransferase gene expression in the endometrium during the pre- and peri-implantation period in cattle

A. M. O'Doherty, L. C. O'Shea, O. Sandra, P. Lonergan, T. Fair and N. Forde

A comprehensive understanding of biological processes in the endometrium is vital for identifying the key factors required for successful embryo recognition and implantation. The aim of the present study was to compare the expression of imprinted genes and DNA methyltransferases in the endometrium of cyclic and pregnant beef heifers. Results from this investigation provide new evidence that, in addition to their role in fetal and placental development, imprinted genes and DNA methyltransferases may have a role in establishing uterine receptivity.

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