Reproduction, Fertility and Development Reproduction, Fertility and Development Society
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 10 2017

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
pp. 1883-1889

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.


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.

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, Khánh X. Nguyễn, Asako Karata, Pilar Ferré, Minh T. Trần, Takuya Wakai and Hiroaki Funahashi
pp. 1902-1909

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.

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

Zina M. Al-Alami, Ziad A. Shraideh and Mutasem O. Taha
pp. 1910-1920

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.

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
pp. 1921-1931

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.

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
pp. 1932-1943

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.


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.

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
pp. 1950-1957

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.

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
pp. 1958-1970

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.

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
pp. 1971-1981

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.

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
pp. 1982-1994

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.

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
pp. 1995-2004

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.

RD16348Mitochondrial ferritin deficiency reduces male fertility in mice

Federica Maccarinelli, Maria Regoni, Fernando Carmona, Maura Poli, Esther G. Meyron-Holtz and Paolo Arosio
pp. 2005-2010

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.

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
pp. 2011-2020

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).


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.

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
pp. 2028-2039

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.

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
pp. 2040-2051

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.

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
pp. 2052-2059

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.


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.

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
pp. 2073-2084

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.

Online Early

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

Published online 18 September 2017

RD17018Expression of kisspeptin protein in hypothalamus and LH profile of growing female lambs

Jolanta Polkowska, Anna Wójcik-G?adysz, Natalia Chmielewska and Marta Wa?kowska
 

Kisspeptin peptide is a major regulator of initiation of pubertal actions in the central nervous system, but its development in growing animals is obscure. The presence of kisspeptin neurons in the brain from birth to puberty in relation to luteinising hormone release pattern was analysed in female lambs. Activity of kisspeptin neurons increased gradually parallel with augmentation of luteinising hormone release and reached maturation several weeks before puberty. The participation of kisspeptin in the development of the ovine reproductive processes is suggested.

Published online 15 September 2017

RD17255Systemic adiponectin treatment reverses polycystic ovary syndrome-like features in an animal model

Anusha Singh, Puran Bora and Amitabh Krishna
 

Adiponectin may be a potent therapeutic agent for recovery of the polycystic ovary syndrome (PCOS) condition. The aim of the present study was to unravel the role of adiponectin in regulating reproduction, metabolism and fertility status in mice in which PCOS had been induced. Reversal of PCOS-like features following adiponectin treatment in mice is a major breakthrough with regard to the effective management of PCOS.

Published online 31 August 2017

RD16486Insulinotropic nucleobindin-2/nesfatin-1 is dynamically expressed in the haemochorial mouse and human placenta

Crystalyn B. Legg-St Pierre, Martina Mackova, Ewa I. Miskiewicz, Denise G. Hemmings, Suraj Unniappan and Daniel J. MacPhee
 

The placenta is a multifunctional link between mother and fetus vital for nutrient transport. In this study we determined when and where nucleobindin-2/nesfatin-1 were located in the placenta and found them particularly in secretory trophoblast cells of the mouse and human placenta. The placenta should be considered a source for nucleobindin-2/nesfatin-1 during pregnancy, with possible roles in maternal glucose control and nutrient sensing.


Some genes are involved in stallion semen tolerance to cryopreservation. This work evaluated the relationship between CRISP-3 protein and some of its single nucleotide polymorphisms, with post-thawing semen quality in stallions. This study demonstrated that different parameters of seminal quality are influenced by the CRISP-3 genotype and the concentration of this protein in seminal plasma.

Published online 30 August 2017

RD17094Milk-fat globule epidermal growth factor 8 (MFGE8) is expressed at the embryo– and fetal–maternal interface in equine pregnancy

S. Barua, A. Macedo, D. S. Kolb, K. E. Wynne-Edwards and C. Klein
 

Milk fat globule-EGF factor 8 protein (MFGE8) is a protein that promotes cell to cell adhesion and is thought to play an essential role in embryo implantation in mice and humans. We have described the expression of MFGE8 in the equine uterus and found particular strong expression of the protein by the fetal portion of the placenta. These results imply a role for MFGE8 in maintaining an intact placenta during mare pregnancy.

Published online 29 August 2017

RD17095Mouse double minute homologue 2 (MDM2) downregulation by miR-661 impairs human endometrial epithelial cell adhesive capacity

Amy Winship, Amanda Ton, Michelle Van Sinderen, Ellen Menkhorst, Katarzyna Rainczuk, Meaghan Griffiths, Carly Cuman and Evdokia Dimitriadis
 

A major limitation of IVF success is failure of the embryo to successfully attach to the womb. We have identified a critical new mechanism of communication between the human embryo and endometrium (inner lining of the uterus). Overall it is likely that this bi-directional communication facilitates the critical adhesion process during the initiation of successful pregnancy.


The testis-specific protein, Y-encoded, 1 (TSPY1) gene is a newly confirmed regulator of sperm production, and dosage deficiency of the multicopy gene confers increased risk of spermatogenic failure. The aim of the present study was to evaluate whether the autosomal homologous genes originating from TSPY1 modify the spermatogenic phenotype of TSPY1. The results suggest that phenotypic expression of TSPY1 defects is independent of variations TSPY-like 1 (TSPYL1) and TSPY-like 5 (TSPYL5), highlighting the significance of TSPY1 in the modulation of spermatogenic efficiency.

Published online 29 August 2017

RD17101MicroRNA expression in bovine preimplantation embryos

Debra K. Berg and Peter L. Pfeffer
 

The types and copy numbers of very short so-called miRNA molecules were measured in eggs and embryos of cattle. Results highlight the importance of a non-canonical miRNA during stages before embryonic transcription begins, the influence of the parental origin of miRNAs and the importance of measuring absolute as opposed to relative levels of these molecules.


Intracytoplasmic sperm injection (ICSI) has been successfully applied to hybrid mice, but not to inbred mice due to low efficiency. Adding an activation step to ICSI of oocytes from juvenile mice results in the birth of healthy inbred pups. The results of this study show that the modified protocol can potentially improve the efficiency of ICSI in inbred mice.

Published online 24 August 2017

RD17157Innate immune genes in persistent mating-induced endometritis in horses

Christina D. Marth, Simon M. Firestone, Dave Hanlon, Lisa Y. Glenton, Glenn F. Browning, Neil D. Young and Natali Krekeler
 

A prolonged inflammatory response to breeding severely decreases fertility in horses. The aim of the present study was to evaluate differences between fertile and subfertile mares in the expression of selected genes associated with the immune response. Three of the genes analysed proved to be very sensitive and specific diagnostic markers predicting prolonged inflammation, and likely reduced fertility, prior to breeding.

Published online 22 August 2017

RD17117Angiogenesis and oxidative stress-related gene variants in recurrent pregnancy loss

Marcela Felix Fortis, Lucas Rosa Fraga, Juliano André Boquett, Thayne Woycinck Kowalski, Caroline Gross Dutra, Rozana Oliveira Gonçalves, Fernanda Sales Luiz Vianna, Lavinia Schüler-Faccini and Maria Teresa Vieira Sanseverino
 

Genetic background can help to explain idiopathic cases of recurrent pregnancy loss (RPL), which make up about 50% of total cases. Here, we have evaluated the genetic profiles of women experiencing RPL by analysing polymorphisms in the NOS2, PTGS2 and VEGFA genes. We found a higher incidence of a polymorphic variant in the NOS2 gene in the RPL group, suggesting a role for this gene in RPL risk, possibly due to oxidative stress mechanisms.

Published online 21 August 2017

RD17037Oral propylene glycol modifies follicular fluid and gene expression profiles in cumulus–oocyte complexes and embryos in feed-restricted heifers

G. Gamarra, C. Ponsart, S. Lacaze, F. Nuttinck, A. Cordova, P. Mermillod, B. Marquant-Le Guienne, D. Monniaux, P. Humblot and A. A. Ponter
 

Ovum pick-up and in vitro embryo production can be used in dairy heifers. However, production results are often disappointing when animals are underfed. A short-term dietary supplement with propylene glycol (400 mL of PG or water/drench, daily drenching at 1600 hours for the first 9 days of the oestrous cycle), given to feed-restricted heifers, was able to restore the expression of selected genes in cumulus cells and blastocysts to levels seen in correctly fed heifers.

Published online 17 August 2017

RD16528Association of CYP19A1 gene polymorphisms with anoestrus in water buffaloes

Khairy M. El-Bayomi, Ayman A. Saleh, Ashraf Awad, Mahmoud S. El-Tarabany, Hadeel S. El-Qaliouby, Mohamed Afifi, Shymaa El-Komy, Walaa M. Essawi, Essam A. Almadaly and Mohammed A. El-Magd
 

One of the predominant causes of anoestrus is a low level of ovarian oestrogens. Herein, we investigated the association of the oestrogen biosynthesis regulator CYP19A1 with the risk of anoestrus in buffaloes and found three SNPs. The risk alleles in these SNPs were significantly associated with lower levels of estradiol and antioxidants and higher incidence of oxidative ovarian damage and anoestrus.


The application of genome storage in conservation is severely constrained by our limited access to reliable assisted breeding techniques. This is particularly so for marsupials, in which the corpus luteum, and thus cycling, is not affected by agents used to control ovarian function in eutherian mammals. Herein we report progress on the use of a single-treatment long-acting suppressor of pituitary function to reset cycling and ovulation in a marsupial.

Published online 08 August 2017

RD17180Can resveratrol attenuate testicular damage in neonatal and adult rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin during gestation?

Rafaela P. Erthal, Gláucia E. M. L. Siervo, Lívia T. R. Silveira, Wellerson R. Scarano and Glaura S. A. Fernandes
 

Humans may be exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an environmental pollutant, via contaminated food, inhalation or skin contact. The effects of TCDD are exerted via binding to aryl hydrocarbon receptors (AhR). In this study we investigated the possible protective effect of resveratrol, an AhR antagonist, against testicular damage caused by TCDD exposure during pregnancy. The findings suggest that TCDD in combination with resveratrol results in more severe toxicity than administration of either drug alone.

Published online 08 August 2017

RD17051Development of fresh and vitrified agouti ovarian tissue after xenografting to ovariectomised severe combined immunodeficiency (SCID) mice

Érica C. G. Praxedes, Gabriela L. Lima, Luana G. P. Bezerra, Fernanda A. Santos, Marcelo B. Bezerra, Denise D. Guerreiro, Ana P. R. Rodrigues, Sheyla F. S. Domingues and Alexandre R. Silva
 

For wild rodents such as the agoutis, the combination of gonadal tissue cryopreservation and xenotransplantation could help promote germplasm conservation and reproduction, especially in the case of genetically valuable individuals. The present study demonstrates that xenografted agouti ovarian tissue, fresh or vitrified, is able to promote the return of ovarian activity in ovariectomised SCID C57B1/6 mice.

Published online 03 August 2017

RD16345Changes in urinary androgen concentration indicate that male giant pandas (Ailuropoda melanoleuca) respond to impending female oestrus during and outside the typical spring breeding season

Barbara L. Gocinski, Katrina K. Knott, Beth M. Roberts, Janine L. Brown, Carrie K. Vance and Andrew J. Kouba
 

Although giant pandas have been described as mono-oestrus spring breeders, males exposed to oestrous females outside this timeframe exhibit breeding behaviours and interest in mating. Urinary androgens and sperm parameters were quantified for males exposed to females expressing oestrus during spring, autumn, or winter. The data from this study showed a rapid physiological readiness of male giant pandas to mate in response to female oestrous cues within or outside of the normal breeding season and may suggest a facultative seasonal reproduction with a “female-induced rut”.

Published online 03 August 2017

RD16494Tissue cell stress response to obesity and its interaction with late gestation diet

Vivek Saroha, Neele S. Dellschaft, Duane H. Keisler, David S. Gardner, Helen Budge, Sylvain P. Sebert and Michael E. Symonds
 

Reduced fetal growth can contribute to a greater risk of later metabolic disease, but the key mechanisms remain unknown. Herein we report findings from a large animal model (sheep) in which fetal growth was restricted by reducing maternal food intake and show that once the offspring become obese, they exhibit increased abundance of molecular markers of metabolic stress in the fat and liver. This could represent a primary mechanism by which growth-restricted offspring are at greater risk of metabolic ill health in later life.

Published online 03 August 2017

RD16487In vitro growth and development of isolated secondary follicles from vitrified caprine ovarian cortex

Érica S. S. Leal, Luis A. Vieira, Naíza A. R. Sá, Gerlane M. Silva, Franciele O. Lunardi, Anna C. A. Ferreira, Cláudio C. Campello, Benner G. Alves, Francielli W. S. Cibin, Johan Smitz, José R. Figueiredo and Ana P. R. Rodrigues
 

In vitro growth of secondary follicles isolated from the vitrified ovarian cortex allows a better understanding of their complex development and of obtaining healthy follicles. The aim of this study was to investigate how the medium composition affects the viability and growth rate of the isolated secondary follicles. This technique could become a strategy to preserve the reproductive potential of females.

Published online 02 August 2017

RD17217The contribution of p53 and Y chromosome long arm genes to regulation of apoptosis in mouse testis

Tomasz Lech, Józefa Styrna and Katarzyna Kotarska
 

Tumor suppressor protein p53, known widely as the guardian of genome preventing cancer formation, is also involved in physiological apoptosis of germ cells in testes. The study used a mouse model to reveal that germ cell apoptosis during the first round of spermatogenesis can be efficiently mediated by p53-independent mechanisms, but activation of these mechanisms requires engagement of multicopy Y-chromosome long arm genes. The role of Y-chromosome genes in regulation of testicular apoptosis was demonstrated for the first time.

Published online 26 July 2017

RD17082Chronic stress effects and their reversibility on the Fallopian tubes and uterus in rats

S. Divyashree and H. N. Yajurvedi
 

It is not known whether female accessory reproductive organs that play a major role in reproduction are vulnerable to chronic stress effects. This study revealed irreversible oxidative stress in the Fallopian tube following short or long-term chronic stress exposure, whereas uterus changes were reversible following short-term exposure but not after long-term exposure. This study indicates that stress-induced reproductive failure may also be due to alterations in the accessory reproductive organs.

Published online 24 July 2017

RD16536Impact of equine assisted reproductive technologies (standard embryo transfer or intracytoplasmic sperm injection (ICSI) with in vitro culture and embryo transfer) on placenta and foal morphometry and placental gene expression

Orlando A. Valenzuela, Anne Couturier-Tarrade, Young-Ho Choi, Marie-Christine Aubrière, Justin Ritthaler, Pascale Chavatte-Palmer and Katrin Hinrichs
 

Assisted reproduction techniques such as in vitro fertilisation and embryo culture have been shown to affect the size and anatomy of the newborn and the placenta in some species, but no information is available on this area in the horse. We evaluated foals and placentas from pregnancies resulting from natural conception, embryo transfer and transfer of in vitro-produced (intracytoplasmic sperm injection) embryos and found no significant differences between groups in size or weight of the foal or placenta and no differences in expression of 17 genes controlling placental function. Our findings suggest that the horse embryo differs from that of ruminants in its sensitivity to the in vitro environment, supporting the continued use of assisted reproduction techniques for commercial foal production.

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

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