Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
Reproduction, Fertility and Development

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.

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