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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 12 2017

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.

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

Giuseppina Basini, Simona Bussolati, Roberta Ciccimarra and Francesca Grasselli
pp. 2305-2312

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.

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
pp. 2313-2323

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.

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
pp. 2324-2335

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.

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
pp. 2336-2344

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.

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
pp. 2345-2356

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.

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.

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
pp. 2367-2375

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.

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.

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
pp. 2387-2399

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.

RD16398Effects of ovarian disaggregation on adult murine follicle yield and viability

Fiona Young, John Drummond, Emma Akers, Louise Bartle, David Kennedy and Mohammad Asaduzzaman
pp. 2400-2410

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.

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
pp. 2411-2418

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.

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
pp. 2419-2429

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.

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
pp. 2430-2436

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.

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
pp. 2437-2445

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.

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
pp. 2446-2456

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.

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
pp. 2457-2465

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.

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
pp. 2466-2478

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.

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
pp. 2479-2495

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.

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
pp. 2496-2505

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

Online Early

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

Published online 22 November 2017

RD16519Composition of marsupial zona pellucida: a molecular and phylogenetic approach

Carla Moros-Nicolás, Pascale Chevret, María José Izquierdo-Rico, William V. Holt, Daniela Esteban-Díaz, Manel López-Béjar, Eva Martínez-Nevado, Maria A. Nilsson, José Ballesta and Manuel Avilés

The zona pellucida (ZP) is an extracellular matrix formed by a few glycoproteins that covers the oocyte and plays an important role during fertilization. The identification of the components of the ZP in marsupials is relevant for the cellular understanding of female gamete formation, sperm interaction with the ZP and fertilisation. Molecular and in silico analyses demonstrated a different ZP composition between Australasian and South American marsupials, being formed from four to six proteins. The present findings have potential relevance from a biological, ecological and economic point of view, related to the development of new contraceptive vaccines for marsupials.

Published online 21 November 2017

RD17096Effect of exogenous progesterone on embryo size and ewe uterine gene expression in an ovine ‘dam size' model of maternal constraint

Lisanne M. Fermin, Sarah J. Pain, Patrick C. H. Morel, Kristene R. Gedye, Paul R. Kenyon and Hugh T. Blair

The preimplantation period of pregnancy is a critical period for determining growth of the conceptus. This study examined the use of progesterone treatment as a means of overcoming maternal constraint effects on embryo growth. Exogenous progesterone was found to indirectly advance embryo growth via alteration of endometrial genes that encode for uterine structural and secretory activity and thereby overcome maternal constraint.

Published online 16 November 2017

RD17102Peri-conception and first trimester diet modifies reproductive development in bulls

K. J. Copping, M. D. Ruiz-Diaz, C. S. Rutland, N. P. Mongan, M. J. Callaghan, I. C. McMillen, R. J. Rodgers and V. E. A. Perry

Nutritional perturbation during early gestation has been shown to alter male reproductive development in rodents and sheep. This study examined the effects of dietary protein perturbation during the peri-conception and first trimester periods upon reproductive development in both fetal and pubertal male bovine progeny. Low maternal dietary protein negatively impacted reproductive development in pubertal and postpubertal offspring. This research highlights that further studies are warranted to explore causal relationships between gestational nutrition and consequent postnatal male reproductive development in cattle.

Published online 15 November 2017

RD17028Molecular cloning and epigenetic change detection of Kiss1 during seasonal reproduction in Chinese indigenous sheep

Xiaoyun He, Qiuyue Liu, Xiaoyu Li, Xiaofei Guo, Xiangyu Wang, Wenping Hu, Ran Di and Mingxing Chu

The Kiss1 gene plays a major role in reproductive seasonality of sheep. This study aimed to confirm that epigenetic modification occurs in sheep during the reproductive cycle. A potential role of histone acetylation modification in altering Kiss1 expression, which may consequently regulate seasonal breeding in sheep, was found. This study may offer a theoretical basis for breeding year-round oestrous sheep.

Published online 15 November 2017

RD17292Proteinaceous sperm motility inhibitory factor from the female Indian garden lizard Calotes versicolor

Goutham Shankar, Shubhashree Uppangala, Satish K. Adiga, Belinda Willard, Bhadravathi K. C. Sagar, Ruth S. K. Titus and Gopal K. Marathe

Female sperm storage is known across many animal phyla, but the mechanism(s) remains elusive. Using the Indian garden lizard, Calotes versicolor as a model, we identified a unique abundant protein of ~55-kDa in the uterovaginal region that controls the motility of the spermatozoa. These studies are likely to unravel the secrets of sperm storage.

Published online 15 November 2017

RD17163Changes in bone turnover and calcium homeostasis during pregnancy and lactation in mammals: a meta-analysis

Diego A. Torres, Mariella B. Freitas and Reggiani V. Gonçalves

Due to fetal and neonatal bone tissue calcification, the maternal calcium demand increases during pregnancy and lactation. To supply this calcium demand, female mammals undergo numerous physiological changes during reproduction. In the present study, we performed a meta-analysis of studies reporting changes in bone mineral density, serum calcium and calciotropic hormone concentrations during pregnancy and lactation in mammals. The results provide a quantitative background to better explain and understand these physiological adaptations.

Published online 10 November 2017

RD17265Effects of N-carbamylglutamate and l-arginine on gonadotrophin-releasing hormone (GnRH) gene expression and secretion in GT1-7 cells

Y. Liu, J. H. Bai, X. L. Xu, Z. L. Chen, L. J. Spicer and T. Feng

N-carbamylglutamate (NCG) and arginine (ARG) supplementation can increase litter size in pigs and rats, but their effects on gonadotrophin-releasing hormone (GnRH) secretion are unknown. We found that NCG and ARG can increase cell numbers and decrease GnRH secretion in murine immortalized GT1-7 cells in vitro by regulating related gene expression. These findings help us to better understand nutritional control of reproduction.

Published online 03 November 2017

RD17354Profilin 1 plays feedback role in actin-mediated polar body extrusion in mouse oocytes

Jun Liu, Qiao-Chu Wang, Xing Duan, Xiang-Shun Cui, Nam-Hyung Kim, Yu Zhang and Shao-Chen Sun

The molecules that are involved in actin-based oocyte polar body extrusion are still largely unknown. In the present study we showed that knockdown of Profilin caused aberrant actin distribution, the failure of polar body extrusion and altered the expression of Rho-associated kinase (ROCK) and myosin light chain (MLC) 2. The results indicate that Profilin is related to ROCK–MLC2 for actin assembly in mouse oocytes.

Published online 25 October 2017

RD17251Seminal plasma and cryopreservation alter ram sperm surface carbohydrates and interactions with neutrophils

Taylor Pini, Tamara Leahy and Simon Paul de Graaf

The reason why some spermatozoa reach the site of fertilisation where others fail is not known, but interaction between spermatozoa and the female environment is key. We profiled ‘winner’ and ‘loser’ spermatozoa and found that the sugars on their surface and their susceptibility to immune cell attack differed significantly. This helps to explain both normal reproductive physiology and the consequences of human intervention.

This research provides new insight into central molecular mechanisms regulating GnRH/LH secretion in anestrous sheep. To better explain this problem, herein, we demonstrated that RF-amide peptides are involved in dopamine-induced inhibition of GnRH/LH secretion. Studies expanded an important issue in the field of neuroendocrinology of reproduction, especially in cognitive appraisals of mechanisms participating in physiological regulation of GnRH/LH biosynthesis.

Published online 11 October 2017

RD17184Expression of vascular endothelial growth factor A isoforms is dysregulated in women with endometriosis

Kevin Danastas, Emily J. Miller, Alison J. Hey-Cunningham, Christopher R. Murphy and Laura A. Lindsay

Angiogenesis is a vital process in the development and survival of lesions during endometriosis. In the present study we identified upregulation of several isoforms of vascular endothelial growth factor (VEGF) A in the endometrium of women with endometriosis during menstruation and, for the first time, identified the natural expression of the VEGF111 isoform in humans. These findings contribute to our understanding of how the endometrium differs in women with endometriosis, and indicates a dysregulation at the critical time of menstruation.

Published online 06 October 2017

RD17356Secretome derived from different cell lines in bovine embryo production in vitro

C. Perrini, P. Esposti, F. Cremonesi and A. Lange Consiglio

Do extracellular vesicles have a biological role in the communication between the embryo and its microenvironment during in vitro culture? In this study we showed that embryos were able to incorporate microvesicles obtained from endometrium and amnion, but only amniotic progenitor cell-derived microvesicles resulted in better blastocyst production rate and quality compared to control. This study could be a useful starting point for further studies related to paracrine mechanisms of communication between embryos and the culture medium.

Published online 04 October 2017

RD17203Effect of melatonin on bovine theca cells in vitro

T. Feng, L. F. Schutz, B. C. Morrell, M. C. Perego and L. J. Spicer

The role of melatonin as an endocrine regulator of reproductive activity is well known; however, whether it stimulates or inhibits reproductive function varies across species. We demonstrated that in vitro exposure to melatonin reduced steroid production by ovarian theca cells in cattle, implicating melatonin as a regulator of reproductive function in cattle. Because reduced reproductive capacity leads to economic losses, an understanding of the effects of melatonin on ovarian function may lead to new ways to increase reproductive efficiency in cattle.

Signalling pathways responsible for follicular activation in the mare remain uncharacterised. Messenger RNA expression and protein localisation of members of the PI3K/AKT and JAK/STAT pathways were analysed using tissue from fetal and adult mare ovaries. Findings suggest that members of these pathways are indeed present and may serve as useful biomarkers for assessment of ovary development in the horse.

Published online 26 September 2017

RD17241Effect of lipopolysaccharide on the expression of inflammatory mRNAs and microRNAs in the mouse oviduct

Katheryn L. Cerny, Rosanne A. C. Ribeiro, Qing Li, James C. Matthews and Phillip J. Bridges

Uncontrolled inflammation of the oviduct can result in tubal occlusion and infertility. This inflammation is often in response to infection with sexually transmitted bacterial pathogens. This study describes how the endotoxin from Escherichia coli-derived lipopolysaccharide affects the expression of inflammatory mRNAs and post-transcriptionally modifying miRNAs within the oviduct, providing novel insight into the molecular regulation of inflammation within this key reproductive organ.

During cryopreservation, spermatozoa undergo cryodamage because of membrane changes, including sterol loss, which results in diminished post-thaw fertilisation ability. The present study showed that loading of cholesterol and desmosterol into chicken spermatozoa prior to cryopreservation enhanced their post-thaw quality by inhibiting apoptotic changes and premature acrosome reaction, providing new mechanistic insight into cryodamage in chicken spermatozoa.

Published online 26 September 2017

RD17049Identification and expression of forkhead box genes in the Chinese giant salamander Andrias davidianus

Qiaomu Hu, Hanbing Xiao, Qilong Wang, Haifeng Tian and Yan Meng

The forkhead box (Fox) gene family encodes multiple forkhead proteins that exhibit marked functional diversity and are involved in biological processes, especially sex differentiation. The identification, characterisation, selective pressure, protein structure and expression pattern of Fox genes in response to temperature were investigated in the present study in the Chinese giant salamander Andrias davidianus to enrich the sexual development and reproduction mechanism in amphibian.

Published online 25 September 2017

RD17209Prospective evaluation of uterine receptivity in mice

Hitomi Nakamura, Takayoshi Hosono, Keiichi Kumasawa and Tadashi Kimura

To improve current infertility treatments, it is necessary to evaluate uterine receptivity in each menstrual cycle. If this can be evaluated beforehand, when uterine receptivity is inadequate eggs could be stored using cryopreservation. In this study in a mouse model, we identified an electrophysiological parameter that can evaluate uterine receptivity prospectively.

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.

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.

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.

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

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 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 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

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 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 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.

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