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Reproduction, Fertility and Development
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RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.


 
 

Reproduction, Fertility and Development publishes research and reviews on all aspects of vertebrate reproduction and developmental biology. More

Editor-in-Chief: Tony Flint 

 
 
 

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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Published online 15 May 2015
DNA methylation reprogramming during oogenesis and interference by reproductive technologies: Studies in mouse and bovine models 
Ellen Anckaert and Trudee Fair

It is estimated that, globally, 56% of couples seek medical care for infertility. Assisted reproductive technologies (ART), such as ovarian stimulation and in vitro embryo production, are used to overcome infertility. Despite increasing use, ART is associated with low pregnancy rates, premature birth and low birth weight. Here we discuss studies investigating the mechanisms by which ART affects oocytes and embryos, paying particular attention to DNA methylation.

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Published online 15 May 2015
Expression profiles of select genes in cumulus–oocyte complexes from young and aged mares 
Lindsay Cox, Dirk K. Vanderwall, Kate C. Parkinson, Alexis Sweat and S. Clay Isom

There is compelling evidence that oocytes from mares >18 years of age have a high incidence of inherent defects that result in early embryonic loss. We compared the gene expression profiles of individual oocytes and cumulus cells from young and aged mares. Three genes (IF3, HSF5 and YBX2) were more abundantly expressed in young mare oocytes and three genes (ARL6IP, BAX and HYOU1) were more abundantly expressed in aged mare cumulus cells. These results confirm there are age-related differences in gene expression in equine cumulus–oocyte complexes, that may be associated with the lower quality and decreased developmental competence of oocytes from aged mares.

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

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

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

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

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

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

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

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

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Published online 08 May 2015
Association of a region of bovine chromosome 1 (BTA1) with age at puberty in Angus bulls 
María E. Fernández, Alberto Prando, Andrés Rogberg-Muñoz, Pilar Peral-García, Andrés Baldo, Guillermo Giovambattista and Juan P. Lirón

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

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Published online 07 May 2015
Effects of age on follicular fluid exosomal microRNAs and granulosa cell transforming growth factor-β signalling during follicle development in the mare 
Juliano C. da Silveira, Quinton A. Winger, Gerrit J. Bouma and Elaine M. Carnevale

Age-related decline in fertility is a consequence of low oocyte number and low oocyte competence (the ability of a fertilised oocyte to develop into an embryo), ultimately resulting in pregnancy failure. Regulation of transforming growth factor (TGF)-β signalling members by small non-coding microRNAs (miRNAs) could explain the age-related effects on oocyte maturation and quality. In the present study we identified altered levels of miRNAs present in follicular fluid exosomes from young and old mares that are predicted to regulate members of the TGF-β signalling pathway in surrounding granulosa cells; this could ultimately help explain age-related decline in fertility.

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

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

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Published online 05 May 2015
Transcriptome of early embryonic invasion at implantation sites in a murine model 
J. M. Moreno-Moya, N. A. Franchi, S. Martínez-Escribano, J. A. Martínez-Conejero, S. Bocca, S. Oehninger and J. A. Horcajadas

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

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

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

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

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

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Published online 05 May 2015
Multiple roles of hypoxia in ovarian function: roles of hypoxia-inducible factor-related and -unrelated signals during the luteal phase 
Ryo Nishimura and Kiyoshi Okuda

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

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Published online 05 May 2015
Relationship between seminal plasma tuberoinfundibular peptide of 39 residues and sperm functional attributes in buffalo (Bubalus bubalis) 
Sellappan Selvaraju, Lakshminarayana Somashekar, Binsila B. Krishnan, Sivashanmugam Parthipan, Guvvala Pushparani, Arunachalam Arangasamy, Duraisamy Rajendran and Janivara Parameswaraiah Ravindra

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

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Published online 05 May 2015
Novel equine conceptus–endometrial interactions on Day 16 of pregnancy based on RNA sequencing 
C. Klein

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

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Published online 01 May 2015
The use of a synthetic progesterone, levonorgestrel (LNG), to control the oestrous cycle in the koala 
K. Ballantyne, S. T. Anderson, M. Pyne, V. Nicolson, A. Mucci, A. Lisle and S. D. Johnston

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

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

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

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Published online 30 April 2015
Comparative genomic identification and expression profiling of a novel ?-defensin gene cluster in the equine reproductive tract 
Gillian P. Johnson, Andrew T. Lloyd, Cliona O'Farrelly, Kieran G. Meade and Sean Fair

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

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Published online 23 April 2015
Centrosome and microtubule functions and dysfunctions in meiosis: implications for age-related infertility and developmental disorders 
Heide Schatten and Qing-Yuan Sun

Chromosomal abnormalities, increased spontaneous abortion and reduced fertility are associated with maternal aging which has resulted in increased demands for assisted reproductive technologies (ARTs) to increase the chances for the birth of a healthy baby. The aim of this study was to review the mechanisms leading to chromosomal abnormalities in aging oocytes with focus on centrosome and microtubule dynamics in the meiotic MII spindle that are affected by maternal aging but can be treated with specific chemicals to delay or reverse the aging effects. Such treatments may find applications for ART procedures in in vitro fertilisation (IVF) clinics to increase the success rates for the birth of healthy babies.

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Published online 22 April 2015
Oogonial stem cells as a model to study age-associated infertility in women 
Neha Garg and David A. Sinclair

This review discusses the link between stem cells, fertility and aging. The work listed provides insight into an emerging field in female fertility, namely, germline stem cells or OSCs and postnatal oogenesis. Given that oocyte numbers decline with age, it is possible that menopause is not a result of exhaustion of the ovarian reserve, rather a result of aging of OSCs.

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Published online 22 April 2015
MicroSort® sperm sorting causes no increase in major malformation rate 
Donald P. Marazzo, David Karabinus, Lawrence A. Johnson and Joseph D. Schulman

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

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Published online 22 April 2015
Spindle configuration and developmental competence of in vitro-matured bovine oocytes exposed to NaCl or sucrose prior to Cryotop vitrification 
Núria Arcarons, Roser Morató, Jozé F. W. Spricigo, Marcia A. M. M. Ferraz and Teresa Mogas

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

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Published online 22 April 2015
Changes in lysosomal enzymes and mannose-6-phosphate receptors related to sexual maturation in bull epididymis 
Andrea C. Aguilera, Lorena Carvelli, Verónica Boschin, Fabián Mohamed, Leila Zyla and Miguel A. Sosa

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

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Published online 21 April 2015
Spatio-specific regulation of endocrine-responsive gene transcription by periovulatory endocrine profiles in the bovine reproductive tract 
Estela R. Araújo, Mariana Sponchiado, Guilherme Pugliesi, Veerle Van Hoeck, Fernando S. Mesquita, Claudia M. B. Membrive and Mario Binelli

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

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Published online 21 April 2015
Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2? and PGE2 production in the bovine endometrium 
Mariko Kuse, Ryosuke Sakumoto and Kiyoshi Okuda

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

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Published online 21 April 2015
Sperm superoxide dismutase is associated with bull fertility 
Kamilah E. Grant, Rodrigo V. de Oliveira, Bettye Sue Hennington, Aruna Govindaraju, Andy Perkins, John Stokes, Dennis Rowe, Einko Topper, Abdullah Kaya, Arlindo Moura and Erdogan Memili

Fertility is the most important factor regulating animal reproduction. The aim of the present study was to ascertain to what extent sperm proteins β-tubulin 2C, heat shock protein 10 and superoxide dismutase 1 (SOD1) were associated with bull fertility using molecular and bioinformatics approaches. We demonstrated that SOD1 was associated with in vivo bull fertility and that it interacts with key regulators of cell viability. The findings are significant because they reveal the paternal effect protein involved in bull fertility.

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

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

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Published online 17 April 2015
Maternal age and in vitro culture affect mitochondrial number and function in equine oocytes and embryos 
W. Karin Hendriks, Silvia Colleoni, Cesare Galli, Damien B. B. P. Paris, Ben Colenbrander, Bernard A. J. Roelen and Tom A. E. Stout

Advancing maternal age and in vitro embryo production (IVP) both predispose to embryonic death. We generated support for the hypothesis that reduced mitochondrial number contributes to compromised embryo quality in these scenarios, by demonstrating reduced mitochondrial DNA (mtDNA) copy numbers in Day 7 embryos from mares older than 12 years, and reduced expression of genes involved in mtDNA replication in oocytes following culture in vitro.

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Published online 17 April 2015
Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques 
J. Dorado, M. J. Gálvez, S. Demyda-Peyrás, I. Ortiz, J. M. Morrell, F. Crespo, J. Gósalvez and M. Hidalgo

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

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Published online 17 April 2015
Expression of leukaemia inhibitory factor at the conceptus–maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare 
M. de Ruijter-Villani, C. Deelen and T. A. E. Stout

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

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Published online 15 April 2015
C-Type natriuretic peptide maintains domestic cat oocytes in meiotic arrest 
Yougang Zhong, Jiabao Lin, Xiaoping Liu, Jian Hou, Yong Zhang and Xingxu Zhao

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

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Published online 15 April 2015
miR-424/322 is downregulated in the semen of patients with severe DNA damage and may regulate sperm DNA damage 
Kai Zhao, Yaoping Chen, Ruifeng Yang, Yang Bai, Cuiling Li, Honggang Li and Chengliang Xiong

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

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Published online 15 April 2015
Potential effects of interferon regulatory factor 4 in a murine model of polyinosinic-polycytidylic acid-induced embryo resorption 
Jing Wang, Tailang Yin, Yanqi Wen, Fuju Tian, Xiaojun He, Danni Zhou, Yi Lin and Jing Yang

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

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Published online 13 April 2015
Hypoxia limits mouse follicle growth in vitro 
J. M. Connolly, M. T. Kane, L. R. Quinlan, P. Dockery and A. C. Hynes

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

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Published online 31 March 2015
Epigenetic pattern changes in prenatal female Sprague-Dawley rats following exposure to androgen 
Yanjie Xia, Shanmei Shen, Xinlin Zhang, Zhantao Deng, Zou Xiang, Hongwei Wang, Long Yi, Qian Gao and Yong Wang

Polycystic ovary syndrome (PCOS), a common cause of female infertility, may be predetermined in the womb. Exposing mothers to excess amounts of an androgenic hormone may render their female offspring susceptible to developing PCOS due to altered regulation of androgen production-related genes in the offspring. This research has furthered our understanding of the mechanisms of PCOS and sheds light on disease prevention.

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Published online 24 March 2015
Cytoskeletal alterations associated with donor age and culture interval for equine oocytes and potential zygotes that failed to cleave after intracytoplasmic sperm injection 
Elena Ruggeri, Keith F. DeLuca, Cesare Galli, Giovanna Lazzari, Jennifer G. DeLuca and Elaine M. Carnevale

Sperm-injected equine oocytes that failed to cleave after ICSI were analysed by confocal microscopy to determine the stage of arrest with emphasis on the cytoskeletal components, actin and tubulin. Alterations associated with cell aging and aging of the oocyte donor were observed, suggesting oocytes from younger mares aged as anticipated with potential attempt at activation, while oocytes from older donors had alterations in actin configurations. Cytoskeletal organisation and zygote development is impacted by maternal and cell aging and is important to understand in clinical applications.

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Published online 19 March 2015
Different locations of RANTES and its receptors on mouse epididymal spermatozoa 
Jin-Hua Wei, Xiao Feng, Zhi-Jian Sun, Pang Cheng, Bin-Fang Ma, Jie Zhao, Yu-Hang Dong, Yuan-Qiang Zhang and Zhen Li

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

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Published online 19 March 2015
Maintaining integrity of germline DNA: individuals age, species do not 
G. E. Seidel, Jr

Individuals age reproductively, partly due to damage to germ-line DNA from environmental insults and errors in DNA replication, but each new generation starts out young. Most problematic germ-line mutations are culled via cell death, nonfunctional gametes, and embryonic death. In addition, degradation at the species level likely is minimised by convergent evolution; no strong evidence was found for rejuvenation of DNA.

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Published online 19 March 2015
Effects of aging on gene expression and mitochondrial DNA in the equine oocyte and follicle cells 
Fernando Campos-Chillon, Todd A. Farmerie, Gerrit J. Bouma, Colin M. Clay and Elaine M. Carnevale

Maternal aging is related to reduced fertility in various species, including the mare. We compared gene expression in follicle cells and oocytes from Young and Old mares after an ovulation-induction signal. Differences were observed in the timing and amount of expression for key regulatory genes between Young and Old mares, suggesting age-associated changes in follicles and oocytes that could impact fertility.

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Published online 19 March 2015
Not just a number: effect of age on fertility, pregnancy and offspring vigour in thoroughbred brood-mares 
Charles F. Scoggin

In thoroughbreds, the influence of age on a mare’s reproductive efficiency and her offspring’s vigour cannot be understated. Age-related changes can contribute to a reduction in fertility and increase the risk of progeny morbidity and mortality. These findings can adversely affect reproductive management of the mare, as well as future commercial value and racing performance of both the mare and her offspring.

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Published online 19 March 2015
The employment of IVF techniques for establishment of sodium, copper and selenium impact upon human sperm quality 
Urszula Marzec-Wróblewska, Piotr Kami?ski, Pawe? ?akota, Marek Szyma?ski, Karolina Wasilow, Grzegorz Ludwikowski, Magdalena Kuligowska-Prusi?ska, Gra?yna Odrow??-Sypniewska, Tomasz Stuczy?ski and Alina Wo?niak

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

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Published online 18 March 2015
Effects of local testicular heat treatment on Leydig cell hyperplasia and testosterone biosynthesis in rat testes 
Zhonghai Li, Jianhai Tian, Genggang Cui, Meng Wang and Dapeng Yu

Rather little attention has been paid to the molecular mechanisms in the effects of heat treatment on Leydig cell function. In this study, testicular heat treatment led to Leydig cell hyperplasia and a reduction in testosterone levels. The findings of this study provide new insights into improvements in the treatment of male infertility.

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    | Supplementary Material (1012 KB)
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Published online 18 March 2015
Activation of P2X7 receptors decreases the proliferation of murine luteal cells 
Jing Wang, Shuangmei Liu, Yijun Nie, Bing Wu, Qin Wu, Miaomiao Song, Min Tang, Li Xiao, Ping Xu, Ximin Tan, Luyin Zhang, Gang Li, Shangdong Liang and Chunping Zhang

Adenosine triphosphate, the energy substrate, can also be an extracellular signalling molecule. The aim of this study was to investigate the role of ATP signalling in murine luteal cells and the possible mechanism(s) involved. We revealed that ATP signalling can activate the P2X7 receptor and inhibited the proliferation of luteal cells.

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Published online 17 March 2015
Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts 
Y. H. Choi, M. C. T. Penedo, P. Daftari, I. C. Velez and K. Hinrichs

Detection of sex, disease status and other genetic traits of horse embryos by biopsy before they are transferred would be a valuable breeding management tool. We found that accuracy differed between methods used for DNA amplification, but not between in vivo- and in vitro-produced embryos. The final high accuracy achieved (>95%) using DNA amplification and multiplex PCR supports clinical use of this technique.

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Published online 17 March 2015
Age-associated changes in granulosa cell transcript abundance in equine preovulatory follicles 
Dawn R. Sessions-Bresnahan and Elaine M. Carnevale

Increased maternal age is associated with reduced pregnancy rates; however, age-related disruptions to normal follicular function remains unknown. The current study determined age-related disruptions to gene regulation in the follicles for many normal processes involved in follicular maturation. We have identified numerous candidate genes that could be used as potential markers for follicle and oocyte viability in aged animals.

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Published online 13 March 2015
Developmental competence of 8–16-cell stage bison embryos produced by interspecies somatic cell nuclear transfer 
L. Antonio González-Grajales, Laura A. Favetta, W. Allan King and Gabriela F. Mastromonaco

Interspecies nuclear transfer can be a valuable tool for the preservation of threatened wildlife species; however, numerous challenges remain. Development of embryos reconstructed from wood bison donor cells transplanted into enucleated domestic cattle oocytes was affected as early as the 8–16-cell stage with altered cellular function and viability. These findings provide evidence for low blastocyst development rates and highlight the influence of interspecies communication in reconstructed embryos.

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Published online 13 March 2015
Sperm rescue in wild African elephants 
Joseph Saragusty, María Teresa Prieto, Alexandre Courtiol, Romain Potier, Frank Göritz, Thomas B. Hildebrandt and Robert Hermes

What do you do when you have large volume of frozen semen from an endangered species with quality too poor to be used in assisted reproductive programs? Facing such a conundrum we tested a number of density-gradient centrifugation media, finding Percoll as the medium of choice for wild African elephant spermatozoa. Rather than discarding semen of inferior quality, density-gradient centrifugation can help select for better-quality and useable spermatozoa.

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Published online 13 March 2015
Relationship between sperm apoptosis and bull fertility: in vivo and in vitro studies 
Lauren Erickson, Tom Kroetsch and Muhammad Anzar

Early assessment of fertility is important for bull selection in a breeding program. The relationships of sperm membrane and DNA fitness with bull fertility were determined. Spermatozoa with membrane and DNA breaks in bull semen adversely affected field fertility following artificial insemination. Cow ova fertilised with semen possessing DNA-nicked spermatozoa demonstrated poor embryonic development in the laboratory.

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Published online 10 March 2015
Human sperm aneuploidy after exposure to polycyclic aromatic hydrocarbons 
Michał Radwan, Joanna Jurewicz, Wojciech Sobala, Sławomir Brzeźnicki, Paweł Radwan, Lucjusz Jakubowski, Wanda Hawuła, Anna Ulańska and Wojciech Hanke

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in daily life and have endocrine-disrupting properties that may affect reproductive and developmental processes. The purpose of this study was to investigate whether 1-hydroxypyrene – a biomarker of PAH exposure – was associated with sperm aneuploidy. The results suggest that exposure to PAHs may be associated with sperm aneuploidy. As this is the first such epidemiological study these findings require further replication.

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Published online 10 March 2015
Steroids in the equine oviduct: synthesis, local concentrations and receptor expression 
Hilde Nelis, Julie Vanden Bussche, Bartosz Wojciechowicz, Anita Franczak, Lynn Vanhaecke, Bart Leemans, Pieter Cornillie, Luc Peelman, Ann Van Soom and Katrien Smits

Early pregnancy influences hormones in the blood of the mare, but little is known about local hormone concentrations in the reproductive tract, which is in direct contact with the developing equine embryo. We determined hormone levels in the oviduct of the horse and our main finding was that the concentration of the pregnancy hormone progesterone is significantly elevated in the oviduct after ovulation and on the side of ovulation. This study provides the basis for further research into embryo–maternal interactions in the equine oviduct.

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    | Supplementary Material (201 KB)
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Published online 05 March 2015
Phenotype switching through epigenetic conversion 
T. A. L. Brevini, G. Pennarossa, S. Maffei and F. Gandolfi

Increasing knowledge of the mechanisms controlling epigenetic regulation of cell fate has provided new understanding of cell commitment and specification. Epigenetic events, such as chromatin remodelling and DNA methylation, have been shown to impose flexible but precise control over the expression of regulatory genes, allowing a dynamic interfacing between genotype and phenotype and modulating cell differentiation plasticity and commitment. In this paper we discuss the use of epigenetic modifiers to obtain high plasticity cells that can then be redirected to acquire a different phenotype. This approach is termed ‘direct epigenetic conversion’. It is a simple, direct and safe way to obtain cells for cell therapy and regenerative medicine.

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Published online 05 March 2015
Antioxidant supplementation during in vitro culture improves mitochondrial function and development of embryos from aged female mice 
Elena Silva, Alison F. Greene, Kevin Strauss, Jason R. Herrick, William B. Schoolcraft and Rebecca L. Krisher

Maternal aging results in reduced blastocyst quality, thought to be due to mitochondrial dysfunction. The present study shows that culture of embryos from aged mice in the presence of a cocktail of antioxidants is beneficial to mitochondrial function and embryo development. Our results highlight the need for a targeted modulation of the redox state to support requirements of embryos originating from reproductively aged females.

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Published online 05 March 2015
The impact of storage temperature and sperm number on the fertility of liquid-stored bull semen 
Craig Murphy, Shauna A. Holden, Edel M. Murphy, Andrew R. Cromie, Patrick Lonergan and Sean Fair

The use of liquid bull semen in artificial insemination provides an ideal opportunity to maximise the number of inseminations per ejaculate through a lower number of spermatozoa per insemination dose. This study demonstrated that a temperature range of 5 to 22°C is not detrimental to liquid sperm function and that lowering the number of spermatozoa per liquid insemination dose from 5 to 3 million does not result in a significant decline in fertility. This reduction in sperm number allows for the better use of genetically superior sires leading to enhanced genetic gain.

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Published online 27 February 2015
Effects of long-term dietary supplementation with conjugated linoleic acid on bovine oocyte lipid profile 
Andrés F. González-Serrano, Christina R. Ferreira, Valentina Pirro, Andrea Lucas-Hahn, Julia Heinzmann, Klaus-Gerd Hadeler, Ulrich Baulain, Patrick Aldag, Ulrich Meyer, Marion Piechotta, Gerhard Jahreis, Sven Dänicke, R. Graham Cooks and Heiner Niemann

Nutritional and environmental conditions around conception and during early embryonic development have significant effects on health and well-being in adult life. Here, we used a bovine heifer model to investigate the effects of rumen-protected fat supplementation on oocyte quality and embryo development with the aid of advanced mass spectrometry and molecular methods. Results show that rumen-protected fatty-acid supplementation affects oocyte lipid content and may pave the way towards a better understanding of reproductive disorders associated with nutritional impairments.

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    | Supplementary Material (753 KB)
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Published online 25 February 2015
Spermatozoa of Sminthopsis murina (Mammalia: Metatheria) exhibit an unusually high degree of chromatin stability in the absence of disulphide bonding in protamine 1 
S. D. Johnston, C. López-Fernández, F. Arroyo, S. Fardell, R. Roy and J. Gosálvez

Despite a lack of cysteine in the protamine fraction of dunnart spermatozoa, the chromatin remains remarkably stable. Using a range of molecular techniques and under conditions known to fragment sperm DNA, we further confirm this phenomenon. Our results suggest that cysteine is present in the histone fraction of the chromatin and it may be functioning to conform tertiary and/or quaternary chromatin structure.

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Published online 24 February 2015
Association between endometrial thickness in oocyte donation cycles and pregnancy success rates 
Hans Arce, Esther Velilla and Marisa López-Teijón

Endometrial receptivity is a primary concern in fertility treatments. No significant differences in pregnancy rate, the number of gestational sacs and miscarriage rates were found with different endometrial thickness measurements in the present study, the largest study so far evaluating the role of endometrial thickness in oocyte donation cycles. Hence, embryo transfer can take place once endometrial thickness >5 mm has been observed in a single ultrasonographic evaluation, with no need for subsequent monitoring.

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Published online 24 February 2015
Oestrous cycle-dependent expression of Fas and Bcl2 family gene products in normal canine endometrium 
S. Henriques, E. Silva, S. Cruz, M. F. Silva, G. Ferreira-Dias, L. Lopes-da-Costa and L. Mateus

Knowledge of the molecular mechanisms regulating apoptosis in normal canine cyclic endometrium might lead to a better understanding of tissue changes associated with implantation and uterine pathological conditions such as cystic endometrial hyperplasia. The results of our work indicate that pro- and anti-apoptotic proteins regulate the balance between cell proliferation and death in the canine endometrium during the oestrous cycle.

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Published online 23 February 2015
Gene expression in the mammary gland of the tammar wallaby during the lactation cycle reveals conserved mechanisms regulating mammalian lactation 
C. J. Vander Jagt, J. C. Whitley, B. G. Cocks and M. E. Goddard

The tammar wallaby has evolved a unique lactation strategy compared to eutherian mammals making it a valuable comparative model for lactation studies. In this study, the tammar wallaby mammary gland was investigated for changes in gene expression during key stages of the lactation cycle and differentially regulated genes were identified. These genes may play a crucial regulatory role in marsupial milk production, and are likely to play a related role in other mammals.

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    | Supplementary Material (2.4 MB)
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Published online 20 February 2015
Maternal control of early embryogenesis in mammals 
Kun Zhang and George W. Smith

Early embryo development in mammals is heavily dependent on the stored maternal factors in oocytes. Herein we summarise the maternal factors affecting early embryo development and functions of maternal-effect genes identified in mammals. Perturbations of maternal control, some of which are associated with ovarian aging, result in decreased oocyte quality.

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Published online 19 February 2015
Expression and immunolocalisation of follicle-stimulating hormone receptors in gonads of newborn and adult female horses 
Dragos Scarlet, Ingrid Walter, Juraj Hlavaty and Christine Aurich

FSH and its receptor (FSHR) are essential for the development and function of the ovary. This study demonstrates for the first time a constant expression and immunolocalisation of FSHR in ovaries from adult mares and newborn fillies, together with the existence of tertiary follicles up to 4 mm in female neonates. Our results suggest an involvement of FSH in luteal function and oocyte maturation in horses.

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Published online 18 February 2015
Use of an organotypic mammalian in vitro follicle growth assay to facilitate female reproductive toxicity screening 
Yuanming Xu, Francesca E. Duncan, Min Xu and Teresa K. Woodruff

Reproductive toxicity screening relies on animal testing. We applied our in vitro follicle growth assay to screen for endocrine and gamete outcomes following exposure to compounds including an oil dispersant used heavily during the Deepwater Horizon oil spill. Here we show that in vitro follicle health can be monitored in vitro, which may help inform future human health exposure policy.

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Published online 18 February 2015
An integrated approach to bovine oocyte quality: from phenotype to genes 
Leslie Angulo, Catherine Guyader-Joly, Sylvain Auclair, Christelle Hennequet-Antier, Pascal Papillier, Mekki Boussaha, Sébastien Fritz, Karine Hugot, François Moreews, Claire Ponsart, Patrice Humblot and Rozenn Dalbies-Tran

In cattle, a suboptimal pool of oocyte factors is suspected to contribute to the poor reproductive performance. Cows with high and low oocyte developmental potential in vitro were discriminated, and differential transcripts evidenced using microarrays; reverse transcription–quantitative polymerase chain reactionconfirmed altered expression for SLC25A16, PPP1R14C, ROBO1, AMDHD1 and MEAF6. These genes will be examined for polymorphisms that could be used in genomic selection.

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    | Supplementary Material (18 KB)
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Published online 18 February 2015
Mutations in the leptin receptor gene associated with delayed onset of puberty are also associated with decreased ovulation and lambing rates in prolific Davisdale sheep 
Jennifer L. Juengel, Michelle C. French, Anne R. O'Connell, Sara J. Edwards, Avijit Haldar, Rudiger Brauning, Phil A. Farquhar, Ken G. Dodds, Susan M. Galloway, Peter D. Johnstone and George H. Davis

Reproductive efficiency is controlled by genetics and environmental factors such as nutrition. Using a sheep model, we have discovered that mutations in the leptin receptor, a key pathway in the nutritional control of reproduction, are associated with reduced fertility. This has potential applications for the selection of sheep with improved reproductive efficiency as well as providing a new model to better understand how leptin regulates fertility in mammals.

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Published online 16 February 2015
Food deprivation and social inequality may lead to oxidative damage: a study on the preventive role of melatonin in the male rat reproductive system 
Shiva Nasiraei-Moghadam, Kazem Parivar, Abolhasan Ahmadiani, Mansoureh Movahhedin and Mohammad-Reza Vaez Mahdavi

Spermatogenic cells are susceptible to oxidative stress. Food deprivation and the sense of inequality in food-deprived rats that see other rats have access to normal food (hereafter referred to as ‘instability’) are known stressors that can increase oxidative stress. In this study, food deprivation and instability increased markers of oxidative stress, whereas melatonin treatment and isolation of rats (which balanced instability) decreased these markers.

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Published online 10 February 2015
Morphological and molecular characterisation of Twitcher mouse spermatogenesis: an update 
Erica Puggioni, Laura Governini, Martina Gori, Giuseppe Belmonte, Paola Piomboni, Elvira Costantino-Ceccarini and Alice Luddi

Altered sphingolipid metabolism has been suggested to cause sperm morphological abnormalities in Twitcher, a mouse model of Krabbe disease. However, our data demonstrate that a derangement of the hypothalamic–pituitary–gonadal axis, caused by severe brain degeneration, may contribute to Twitcher infertility. These findings may also contribute in further understanding of all cases of infertility associated with neurological impairment.

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Published online 09 February 2015
Effects of ovarian hyperstimulation on mitochondria in oocytes and early embryos 
Jing Shu, Li-Li Xing, Guo-Lian Ding, Xin-Mei Liu, Qing-Feng Yan and He-Feng Huang

Millions of babies come from artificial reproductive technology, which harvests eggs commonly by superovulation. Is this safe? We studied the superovulation effects on mice egg mitochondria, the important organelle reflecting egg potential and with genetic function. The results showed a decline in the average mitochondrial quality by superovulation. However, it is not superovulation that causes the damage, it is superovulation that recruits more eggs with poor mitochondrial quality instead.

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Published online 14 January 2015
Frequency of exogenous hormone therapy impacts spermiation in male Fowler 
Caitlin E. McDonough, Michelle W. Martin, Carrie K. Vance, Judith A. Cole and Andrew J. Kouba

Amphibians are experiencing a global extinction crisis and captive assurance colonies in zoos and aquariums are unsustainable over the long-term due to poor reproduction. In this study, we aimed to improve our understanding of the use of hormone therapy in male toads for sperm collection. Our results have clear implications that frequent hormone-induced spermiation could have a negative effect on sperm production and we recommend that hormone injections be spaced a minimum of 2 weeks apart to optimise the health of the animals and provide optimum sperm quality for assisted reproductive technologies.

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Published online 14 January 2015
The effects of glucuronic acid and N-acetyl-D-glucosamine on in vitro fertilisation of porcine oocytes 
K. Schmidt, A. Clark, A. Mello, C. Durfey, A. Buck, K. Boyd and B. D. Whitaker

Reducing the incidence of polyspermic penetration may aid in improving embryonic development and further the understanding of fertilisation in pigs. This study reveals that by supplementing oocytes with perivitelline space components, the space thickness increases and polyspermic penetration is reduced. The findings highlight the need for adequate environments to improve the maturation of oocytes prior to fertilisation.

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Published online 13 January 2015
Maternal treatment with fluoxetine promotes testicular alteration in male rat pups 
Aline C. Ramos, Alice H. dos Santos, Kennia M. Silveira, Ana Carolina I. Kiss, Suzana F. P. Mesquita and Daniela C. C. Gerardin

Previous results obtained in our laboratory showed that maternal exposure to fluoxetine (FLX) alters testicular morphometric parameters in adult male offspring. The present study investigated the effects of maternal FLX exposure on maternal behavior and on testes function offspring in the weanling and pubertal periods in rats. FLX treatment did not impair mother–pup interactions; however, it could impair testicular function in offspring.

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Published online 13 January 2015
Genetic influence on the reduction in bovine embryo lipid content by l-carnitine 
Luis Baldoceda, Dominic Gagné, Christina Ramires Ferreira and Claude Robert

Holstein and Jersey embryos display difference in term of lipid profile and molecular characteristics, suggesting a variance in lipid metabolism. Using L-carnitine, we noticed a reduction in embryonic lipid content for both breeds, but the response to treatment is weaker in the Jersey. L-carnitine supplementation reduces lipid content of blastomere and offers the potential to improve cryopreservation success.

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Published online 07 January 2015
Characterisation of several ankyrin repeat protein variant 2, a phosphoprotein phosphatase 1-interacting protein, in testis and spermatozoa 
Joana Vieira Silva, Luís Korrodi-Gregório, Georg Luers, Maria João Cardoso, António Patrício, Nuno Maia, Edgar F. da Cruz e Silva and Margarida Fardilha

Phosphoprotein phosphatase 1 (PPP1) is essential for male fertility and the key to understanding PPP1 function lies in the identification of its interacting partners. This study was undertaken to characterise several ankyrin repeat protein variant 2 (SARP2), a PPP1-interacting protein, highly enriched in testis and spermatozoa. Given the expression pattern of SARP2 and its association with PPP1 and sperm-associated antigen 9, we suggest that it plays a role in spermiogenesis and sperm function, namely in sperm motility and the acrosome reaction.

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Published online 07 January 2015
Expression and function of transcription factor AP-2γ in early embryonic development of porcine parthenotes 
Sung-Hyun Lee, Jung-Woo Kwon, Inchul Choi and Nam-Hyung Kim

This is the first study to report on the expression and functions of transcription factor activator protein (AP) 2γ in pig embryos. AP2γ was detected at all stages during preimplantation and depletion negatively affected early embryo development. These findings suggest that AP2γ may play a role in critical cellular events, including cell adhesion, proliferation and cell fate, during preimplantation development.

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Published online 07 January 2015
Prenatally administered dexamethasone impairs folliculogenesis in spiny mouse offspring 
Monika Hułas-Stasiak, Piotr Dobrowolski and Ewa Tomaszewska

Prenatal exposure to dexamethasone causes developmental alterations in the ovaries of offspring. This study demonstrates that dexamethasone administered during the fetal period enhances follicular atresia by activating programmed cell death, which, in turn, depletes the stock of ovarian follicles and can lead to reduction of fertility in adult females. The findings highlight the need for careful use of dexamethasone in pregnant women.

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Published online 07 January 2015
Gelatinases and their tissue inhibitors are associated with oxidative stress: a potential set of markers connected with male infertility 
Ewa M. Kratz, Anna Kałuża, Mirosława Ferens-Sieczkowska, Beata Olejnik, Renata Fiutek, Mariusz Zimmer and Agnieszka Piwowar

The search for new diagnostic markers of male infertility takes into account factors other than sperm characteristics. The expression of matrix metalloproteinase (MMP)-9 and MMP-2 may be regulated by oxidative stress in various pathophysiological processes. Thus, the presence of seminal MMPs, their tissue inhibitors (tissue-specific inhibitor of metalloproteinase (TIMP)-1, TIMP-2) and their correlation with levels of oxidative stress parameters (i.e. advanced oxidation protein products (AOPP)) is interesting in the context of male fertility. High AOPP and an increased MMP-9 : TIMP-1 ratio alters the oxidative–antioxidative balance of the ejaculate, thereby reducing male fertility. Thus, reducing male fertility, and therefore these parameters may serve as additional diagnostic markers of semen quality and male reproductive potential.

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Published online 07 January 2015
Cooled semen for fixed-time artificial insemination in beef cattle 
Juliana C. Borges-Silva, Márcio R. Silva, Daniel B. Marinho, Eriklis Nogueira, Deiler C. Sampaio, Luiz Orcírio F. Oliveira, Urbano G. P. Abreu, Gerson B. Mourão and Roberto Sartori

Oestradiol-based fixed-time artificial insemination (FTAI) programs have a relatively wide window of time for ovulation that may decrease pregnancy per AI (P AI–1) depending on semen processing. We have shown a 20% increase in P AI–1 in beef cattle when cooled versus frozen–thawed semen was used, because of better sperm quality and longevity.

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Published online 07 January 2015
Increased mRNA expression of selected pro-inflammatory factors in inflamed bovine endometrium in vivo as well as in endometrial epithelial cells exposed to Bacillus pumilus in vitro 
Martina A. Gärtner, Sarah Peter, Markus Jung, Marc Drillich, Ralf Einspanier and Christoph Gabler

Inflammatory processes in the bovine endometrium caused by bacterial infection often result in reduced fertility. We aimed to elucidate the first inflammatory response of the bovine endometrium. Increased mRNA expression of pro-inflammatory factors was connected to inflammation in vivo and was provoked in endometrial epithelial cells co-cultured with the potentially pathogenic Bacillus pumilus. These findings suggest that the endometrium can react promptly to bacterial infections.

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Published online 07 January 2015
Histochemical structure and immunolocalisation of the hyaluronan system in the dromedary oviduct 
Omnia Mohey-Elsaeed, Waleed F. A. Marei, Ali A. Fouladi-Nashta and Abdel-Aleem A. El-Saba

Hyaluronic acid is a major component of the extracellular matrix and is also found in body fluids. We show immunohistochemical localization of its synthases, main receptor and degrading enzymes in the camel oviduct. Regional differences in its synthesis and degradation suggest a differential role in the oviduct–gamete interaction in the ampulla and isthmus.

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Published online 05 January 2015
Actin crosslinking protein filamin A during early pregnancy in the rat uterus 
Romanthi J. Madawala, Connie E. Poon, Samson N. Dowland and Christopher R. Murphy

Filamin A is an actin-binding protein that was found to be associated with actin cytoskeletal remodelling of both luminal uterine epithelial cells (UECs) and endometrial stromal cells (ESCs) during early pregnancy. The distribution of filamin A in the endometrium was influenced by deciduogenic factors, while distribution of filamin A was found to be dependent on ovarian hormones. The expression of filamin A in the endometrium during early pregnancy contributes to the molecular and morphological changes that both UECS and ESCs undergo in order for them to be receptive to implantation by contributing to the remodelling of the actin cytoskeleton and hence the maintenance of plasma membrane stability of UECs and ESCs during early pregnancy.

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Published online 05 January 2015
Creation of DNA aptamers against recombinant bone morphogenetic protein 15 
Jun Sheng Lin, Alexia Kauff, Yong Diao, Huiyong Yang, Steve Lawrence and Jennifer L. Juengel

The oocyte-derived growth factor bone morphogenetic protein (BMP) 15 plays important roles in fertility, but its mechanism of action remains poorly understood. Sequences of single-stranded DNA specifically recognising BMP15 have been generated by a directed molecular evolutionary process. These special sequences (aptamers) can be used as a tool to detect BMP15 and decipher its biological actions.

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Published online 05 January 2015
Effects of dietary quercetin on female fertility in mice: implication of transglutaminase 2 
Kelly E. Beazley and Maria Nurminskaya

Increasing use of dietary supplementation with bioflavonoids such as quercetin raises concerns about their potential impacts on reproductive health. Here we show a negative impact of quercetin consumption on female fertility in the animal model and identify its target, transglutaminase 2, as a novel regulator of ovarian ageing. Thus the beneficial effects of quercetin may be overshadowed in females of reproductive age.

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Published online 17 December 2014
High oxygen tension increases global methylation in bovine 4-cell embryos and blastocysts but does not affect general retrotransposon expression 
Wenwen Li, Karen Goossens, Mario Van Poucke, Katrien Forier, Kevin Braeckmans, Ann Van Soom and Luc J. Peelman

Environmental stresses can have detrimental effects on the developing embryo that resonate throughout the rest of the life of that individual. In this study we tested whether the negative effects are induced by changes the chromatin structure and, if so, whether this loosens the brake on the activity of retrotransposons. Although global chromatin changes were observed, the results indicate that retrotransposons are also held in check by other factors.

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    | Supplementary Material (409 KB)
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Published online 17 December 2014
New evidence of melatonin receptor contribution to ram sperm functionality 
Marta Gonzalez-Arto, Caorlina Luna, Rosaura Pérez-Pé, Teresa Muiño-Blanco, José A. Cebrián-Pérez and Adriana Casao

This study analysed the effects of melatonin through its receptors (MT1 and MT2) on ram sperm functionality. Using immunofluorescence assays and by evaluating the effects of MT1 and MT2 receptor agonists and antagonists, we determined that the modulating effect of melatonin on ram sperm capacitation seems to be receptor mediated, primarily via MT2 receptors. These results may explain the effects of exogenous melatonin on ram fertility.

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Published online 17 December 2014
Luteotropic and luteolytic factors regulate mRNA and protein expression of progesterone receptor isoforms A and B in the bovine endometrium 
Robert Rekawiecki, Magdalena Karolina Kowalik and Jan Kotwica

Expression of nuclear progesterone receptor isoforms A and B (PGRA and PGRB, respectively) can be regulated by various factors. PGRA and PGRB mRNA and protein expression was determined in bovine endometrium after stimulation with selected luteotropic and luteolytic factors. The data suggest that these factors affect PGRA and PGRB mRNA and protein levels and, in this way, regulate the effects of progesterone on endometrial cells.

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Published online 17 December 2014
Nutrition affects Sertoli cell function but not Sertoli cell numbers in sexually mature male sheep 
Yongjuan Guan, Guanxiang Liang, Penny A. R. Hawken, Sarah J. Meachem, Irek A. Malecki, Seungmin Ham, Tom Stewart, Le Luo Guan and Graeme B. Martin

A comprehensive evaluation of Sertoli cells has been used to investigate a reversible, non-pathological change in testis mass induced by nutrition in mature sheep. Under-nutrition did not affect Sertoli cell number but did affect Sertoli cell function, including reversal of cellular differentiation and disruption of tight-junctions. These responses were associated with major decreases in the output of spermatozoa.

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Published online 17 December 2014
Embryonic developmental plasticity in the long-snouted seahorse (Hippocampus reidi, Ginsburg 1933) in relation to parental preconception diet 
Francisco Otero-Ferrer, Marisol Izquierdo, Alireza Fazeli and William V. Holt

In seahorses, embryo development occurs within the male instead of the female, leading us to ask whether the male’s brood pouch has a similar function to the mammalian placenta. We specifically investigated whether the male’s diet before conception can influence the future survival and well being of newborn seahorses. Our experiment confirmed that to optimise the health and future survival of seahorses, male parents need to be supplied with dietary unsaturated fatty acids before they become pregnant.

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Published online 17 December 2014
Impact of superovulation and mating on the wellbeing of juvenile and adult C57BL/6N mice 
Thomas Kolbe, Sarjoun Sheety, Ingrid Walter, Rupert Palme and Thomas Rülicke

Superovulation of mice is preferentially applied before sexual maturity of females is reached in order to maximize ova production. The effect of donor age in mice was evaluated with regard to distress induced by hormonal treatment and mating. Our results confirmed higher ovulation rates for immature females compared to adults and revealed no significant welfare issues for juvenile mice used for superovulation.

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Published online 08 December 2014
Prospects for immunocontraception in feral horse population control: exploring novel targets for an equine fertility vaccine 
Aleona Swegen and R. John Aitken

Rapidly growing populations of feral horses can cause significant damage to landscapes and rural economies. Animal welfare concerns along with social and ethical considerations are increasingly precluding the use of lethal control methods to maintain acceptable population sizes, while non-lethal fertility control is not yet able to provide effective alternatives. This review explores the unique aspects of equine reproductive physiology that may provide opportunity for developing novel fertility vaccine strategies.

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Published online 08 December 2014
Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis 
Lalitha Devi, Himesh Makala, Lavanya Pothana, Khemlal Nirmalkar and Sandeep Goel

Testis cryopreservation is an efficient tool for preservation of the genetic potential of immature males that die before attaining sexual maturity. This is especially useful for buffalo calves that suffer a high mortality rate. Our results show that an optimum concentration of dimethylsulfoxide and fetal bovine serum could effectively preserve immature buffalo testicular tissues subjected to an uncontrolled slow-freezing protocol. Our findings encourage the cryo-banking of immature buffalo testicular tissues.

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Published online 04 December 2014
Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification 
Roser Morató, Míriam Castillo-Martín, Marc Yeste and Sergi Bonet

The ability to correctly select viable embryos is relevant to improving pregnancy rates and avoiding futile transfer attempts. Herein, we assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity prior to vitrification. Faster embryo development underlies higher blastocyst cryotolerance. The best results for survival after warming were obtained with the vitrification of porcine blastocysts presenting a high degree of expansion, particularly when vitrification was performed after 6 days of in vitro embryo culture .

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Published online 03 December 2014
Controlled hydrostatic pressure stress downregulates the expression of ribosomal genes in preimplantation embryos: a possible protection mechanism? 
I. Bock, H. Raveh-Amit, E. Losonczi, A. C. Carstea, A. Feher, K. Mashayekhi, S. Matyas, A. Dinnyes and C. Pribenszky

Stress tolerance of gametes and embryos can be increased by preconditioning with hydrostatic pressure (HP) treatment. To study the effect of HP, mouse oocytes were treated, fertilised and then analysed by gene expression microarray at the 4-cell stage. The results revealed that numerous ribosomal genes were downregulated after HP treatment, suggesting that regulation of translation has a major role in hydrostatic pressure-induced stress tolerance.

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    | Supplementary Material (14 KB)
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Published online 03 December 2014
Sexual maturation of the Mongolian gerbil (Meriones unguiculatus): a histological, hormonal and spermatic evaluation 
Maria Etelvina Pinto-Fochi, Ana Carolina Negrin, Wellerson Rodrigo Scarano, Sebastião Roberto Taboga and Rejane Maira Góes

Several reproductive aspects of Mongolian gerbil still remain unknown. This study determined the phases of sexual development of this rodent based on an integrative analysis of testicular morphology, hormonal data and sperm parameters. In terms of laboratory rodents, the gerbil has a similar reproductive profile with the rat. These reproductive data provide new foundations for future investigations involving toxicological or developmental studies with this species.

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Published online 02 December 2014
Cellular changes in the hamster testicular interstitium with ageing and after exposure to short photoperiod 
E. Beltrán-Frutos, V. Seco-Rovira, C. Ferrer, J. F. Madrid, F. J. Sáez, M. Canteras and L. M. Pastor

The testicular interstitium is responsible for endocrine function in the testis. We have studied the cellular changes in two physiological processes of testicular involution, one reversible due to exposure to short photoperiod and the other irreversible due to ageing. These processes induce very different changes in the cells. These differences are probably related to cell turnover in the testicular interstitium and to the non-apoptotic but necrotic character of Leydig cell death.

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Published online 28 November 2014
Involvement of prolactin in the meloxicam-dependent inflammatory response of the gonadotropic axis to prolonged lipopolysaccharide treatment in anoestrous ewes 
Andrzej P. Herman, Agata Krawczyńska, Joanna Bochenek, Hanna Antushevich, Anna Herman and Dorota Tomaszewska-Zaremba

An immune challenge can affect the reproductive process in females. In this study, performed in ewes, we demonstrated that inhibition of cyclo-oxygenase 2 by meloxicam could minimise the negative effect of systemic inflammation induced by seven injections of bacterial endotoxin on gonadotropin-releasing hormone and LH secretion. However, the effectiveness of meloxicam treatment may be reduced during an inflammatory state characterized by high levels of prolactin.

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Published online 27 November 2014
Localisation and endocrine control of hyaluronan synthase (HAS) 2, HAS3 and CD44 expression in sheep granulosa cells 
R. Chavoshinejad, W. F. A. Marei, G. M. Hartshorne and A. A. Fouladi-Nashta

We localised hyaluronan (HA) synthases (HAS) 1–3 and their main receptor CD44 in sheep antral follicles, showing intense staining in granulosa cells. Analyses of hormonal regulation of the HA system in cultured sheep granulosa cells indicated differential regulation of HAS2, HAS3 and CD44 expression by oestradiol, insulin-like growth factor, insulin and gonadotropins.

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Published online 27 November 2014
Stable reference genes in granulosa cells of bovine dominant follicles during follicular growth, FSH stimulation and maternal aging 
Muhammad Irfan-ur-Rehman Khan, Fernanda Caminha Faustino Dias, Isabelle Dufort, Vikram Misra, Marc-Andre Sirard and Jaswant Singh

Little is known about stability of reference genes during dominant follicle development. This study examined the expression of a number of granulosa cell genes during ovarian superstimulation and maternal aging in cattle. Results support the use of the geometric mean of four genes (UBE2D2, EIF2B2, GAPDH and SF3A1) for relative gene expression analyses of granulosa cells during maternal and follicular aging.

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Published online 26 November 2014
Taking control of the female fertile lifespan: a key role for Bcl-2 family proteins 
Seng H. Liew, Kavitha Vaithiyanathan and Karla J. Hutt

The number of eggs laid down in the ovary at birth and subsequently maintained during reproductive life influences the length of the female fertile lifespan. Recent studies have revealed a critical role for cell death regulators belonging to the BCL-2 protein family in determining how many eggs are available to females to support fertility. This review summarises our current understanding of the proteins responsible for initiating and mediating cell death during the establishment and maintenance of the female germline.

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Published online 25 November 2014
Suppressed expression of macrophage migration inhibitory factor in the oviducts of lean and obese cows 
Asrafun Nahar and Hiroya Kadokawa

Oviducts synthesise macrophage migration inhibitory factor (MIF) to promote sperm capacitation and embryogenesis. We discovered that both obese and lean Japanese Black cows have suppressed expression of MIF mRNA and protein in the ampullae and isthmuses of their oviducts. Therefore, maternal nutritional condition may change expression of oviductal MIF, which plays important roles in fertilisation and early embryo development.

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Published online 21 November 2014
Association of C677T transition of the human methylenetetrahydrofolate reductase (MTHFR) gene with male infertility 
Mohammad Karimian and Abasalt Hosseinzadeh Colagar

The human methylenetetrahydrofolate reductase (MTHFR) gene, located at chromosome 1p36.3, is associated with male infertility. Previous reports reported that the common polymorphisms C677T, A1298C and G1793A may be related to infertility. This study shows that the C677T transition plays an important role in the change in affinity of the MTHFR–Flavin adenine dinucleotide binding site, increasing the risk of infertility.

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Published online 20 November 2014
Prepubertal male rats with high rates of germ-cell apoptosis present exacerbated rates of germ-cell apoptosis after serotonin depletion 
Néstor Méndez Palacios, María Elena Ayala Escobar, Maximino Méndez Mendoza, Rubén Huerta Crispín, Octavio Guerrero Andrade, Javier Hernández Melández and Andrés Aragón Martínez

Genetic expression could be behind individual sensitivity to chemical substances. Here we show that individuals with innate low or high levels of germ-cell death have distinct gene expression and that levels of germ-cell death changed differentially after administration of amphetamine. This helps both to understand the exacerbated response to drugs by some individuals and to develop a rationale in individualised medicine.

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    | Supplementary Material (14 KB)
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Published online 20 November 2014
Effect of spontaneous term labour on the expression of the NR4A receptors nuclear receptor related 1 protein (Nurr1), neuron-derived clone 77 (Nur77) and neuron-derived orphan receptor 1 (NOR1) in human fetal membranes and myometrium 
Martha Lappas

The upstream mechanisms involved in the processes of human labour are yet to be fully elucidated. This study shows that members of the nuclear receptor subfamily 4A (NR4A) are increased in fetal membranes and myometrium obtained from women after spontaneous term labour onset, and in the presence of various bacterial products. These findings suggest that NR4A family members may play a role in the processes of human labour including fetal membrane rupture and myometrial contractions.

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Published online 20 November 2014
Treatment of buffalo (Bubalus bubalis) donor cells with trichostatin A and 5-aza-2 
M. Saini, N. L. Selokar, H. Agrawal, S. K. Singla, M. S. Chauhan, R. S. Manik and P. Palta

Skin cells are widely used as nuclear donors for producing cloned animals. We showed that treatment of these cells with certain chemical agents, which affect their DNA and the proteins associated with it, alters the characteristics and improves the clonability of these cells. This could help to improve the yield and quality of cloned embryos derived from skin cells.

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    | Supplementary Material (986 KB)
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Published online 19 November 2014
Association of decreased spermatozoa omega-3 fatty acid levels and increased oxidative DNA damage with varicocele in infertile men: a case control study 
Li-Xin Tang, Dong-Juan Yuan, Qi-Ling Wang, Fang Jiang, Jian Guo, Yun-Ge Tang, Li-Xin Zheng and Jing X. Kang

Identifying the abnormalities in spermatozoa of infertile men may aid in the development of new interventions for infertility. This study reveals that infertile men with varicocele have lower omega-3 fatty acids levels and greater oxidative DNA damage in their spermatozoa. The findings highlight the need for intervention trials to test the usefulness of omega-3 supplementation in reducing sperm abnormalities in infertile men with varicocele.

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Published online 17 November 2014
CXADR-like membrane protein (CLMP) in the rat ovary: stimulation by human chorionic gonadotrophin during the periovulatory period 
Feixue Li, Xiaoping Miao, Yonglong Chen and Thomas E. Curry

Cell–cell adhesion is important for cellular communication, cellular structure and steroidogenesis. The present study demonstrates that an ovulatory stimulus of human chorionic gonadotrophin (hCG) stimulates ovarian expression of the cell adhesion molecule CXADR-like membrane protein (CLMP). The hCG induction of Clmp mRNA is mediated through the protein kinase A signalling pathway by activation of the epidermal growth factor receptor and requires the synthesis of new protein. The temporal stimulation of ovarian CLMP may be important in cell adhesion events during ovulation and luteal formation.

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Published online 17 November 2014
Neonatal testicular cell transplantation restores murine spermatogenesis damaged in the course of herpes simplex virus-induced orchitis 
Ekaterina A. Malolina, Andrey Yu. Kulibin and Alla A. Kushch

There is growing evidence that viral infections of the genital tract may contribute to male infertility and cause orchitis. The present study was carried out to develop cell therapy for this disorder. We revealed that a combination of antiviral therapy and neonatal testis-cell transplantation is effective for restoration of murine spermatogenesis damaged by herpes simplex virus infection.

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Published online 13 November 2014
N-Acetylcysteine improves oocyte and embryo quality in polycystic ovary syndrome patients undergoing intracytoplasmic sperm injection: an alternative to metformin 
Ebrahim Cheraghi, Malek Soleimani Mehranjani, Mohammad Ali Shariatzadeh, Mohammad Hossein Nasr Esfahani and Zahra Ebrahimi

Although the number of oocytes in patients with polycystic ovary syndrome (PCOS) is usually greater than normal following ovulation stimulation, these oocytes are often of low quality. The aim of the present study was to investigate the effects of metformin and N-acetylcysteine (NAC) on follicular fluid parameters, as well as oocyte and embryo quality, in Iranian women with PCOS. We conclude that NAC improves oocyte and embryo quality, and could be administered as an alternative to metformin.

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    | Supplementary Material (132 KB)
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Published online 13 November 2014
Leukotriene production profiles and actions in the bovine endometrium during the oestrous cycle 
Anna J. Korzekwa, Robert Milewski, Martyna Łupicka and Dariusz J. Skarzynski

Leukotrienes are involved in reproductive functions in cows; however, their particular role in endometrium is unknown. The aim of the study was to elucidate the actions of leukotriene B4 and C4 in the bovine uterus. We demonstrated that these eicosanoids are produced, secreted and act in the endometrium depending upon the luteal stage. The results indicate multiple actions of leukotrienes in the bovine reproductive tract.

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Published online 11 November 2014
Inhibition of angiopoietin-1 (ANGPT1) affects vascular integrity in ovarian hyperstimulation syndrome (OHSS) 
Leopoldina Scotti, Dalhia Abramovich, Natalia Pascuali, Luis Haro Durand, Griselda Irusta, Ignacio de Zúñiga, Marta Tesone and Fernanda Parborell

OHSS is a disorder associated with ovarian stimulation and is characterised by ovarian enlargement and vascular permeability. The aim was to investigate the effect of follicular fluid from women at risk of OHSS, in the presence of an ANGPT1 inhibitor, on the chorioallantoic membrane assay and in endothelial cell culture. Our findings suggest that ANGPT1 increases pathophysiological angiogenesis in patients at risk of OHSS.

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Published online 07 November 2014
Ovarian acyclicity in zoo African elephants (Loxodonta africana) is associated with high body condition scores and elevated serum insulin and leptin 
Kari A. Morfeld and Janine L. Brown

There is growing concern that obesity is negatively impacting health of many species – including the world’s largest land mammal – the elephant. This study provides the first evidence that poor reproduction in zoo African elephants is related to excessive body fat and altered levels of obesity-related hormones. Promoting weight loss in non-cycling females with the goal of improving metabolic health and resumption of ovarian activity may combat the non-sustainability of zoo African elephant populations.

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Published online 07 November 2014
Influences of melatonin treatment, melatonin receptor 1A (MTNR1A) and kisspeptin (KiSS-1) gene polymorphisms on first conception in Sarda ewe lambs 
S. Luridiana, M. C. Mura, C. Daga, G. Cosso, S. Bodano, F. Farci, F. Zidda and V. Carcangiu

Many factors are involved in the onset of reproductive activity in sheep, such as melatonin, its own receptor gene (MTNR1A) and kisspeptin (KiSS-1) gene. The aim of this study was to investigate if melatonin implants and MTNR1A and KiSS-1 gene polymorphisms can advance the first conception; we found that the G/G genotype of the MTNR1A gene improved the reproductive response to melatonin treatment. Understanding the mechanisms of melatonin function may improve reproductive performance in sheep.

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Published online 05 November 2014
Endothelin as a local regulating factor in the bovine oviduct 
Yuki Yamamoto, Misa Kohka, Yoshihiko Kobayashi, Izabela Woclawek-Potocka and Kiyoshi Okuda

Endothelin regulates oviductal motility, which is important for the transport of gametes and embryo. To define the endothelin system in the bovine oviduct, the production and target sites of endothelin were evaluated. All isoforms of endothelin are produced by epithelial cells and endothelin receptors localise in the smooth muscle, suggesting that the bovine oviduct possesses a local endothelin system.

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Published online 31 October 2014
Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality 
Noelia Prieto-Martínez, Ingrid Vilagran, Roser Morató, Joan E. Rodríguez-Gil, Marc Yeste and Sergi Bonet

The aquaporin protein family allows the transport of water and other solutes through cell membranes and plays a role in reproductive physiology. The present work has studied, for the first time, the presence of AQP7 and AQP11 in boar spermatozoa. Although both were identified in this species, only AQP11 was found to be related to sperm quality.

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Published online 31 October 2014
Aberrant expression of TAR DNA binding protein-43 is associated with spermatogenic disorders in men 
Divya Saro Varghese, Uma Chandran, Ambili Soumya, Sathy M. Pillai, Krishnapillai Jayakrishnan, Prabhakara P. Reddi and Pradeep G. Kumar

Defects in the molecular mechanisms involved in the production and normal functioning of spermatozoa could be a major cause of male factor infertility. We identified a strong association between the absence of a critical molecule in testicular germ cells and male factor infertility. Identification of such critical molecules and their subsequent use in diagnosing male factor infertility may improve infertility management strategies.

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    | Supplementary Material (937 KB)
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Published online 30 October 2014
Proteome profiling of the sperm maturation milieu in the rhesus monkey (Macaca mulatta) epididymis 
Xin Liu, Shao-Hua Jin, Xue-Xia Liu, Wen-Juan Wang and Fu-Jun Liu

The first gel-based profiling of the rhesus monkey (Macaca mulatta) sperm-milieu proteome was established by a two-dimensional electrophoresis map. When compared with bull and human sperm-milieu proteins, those in the rhesus monkey had a close evolutionary relationship to human. It could be used as an ideal model for the development of diagnostic and therapeutic targets for infertility or contraception.

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Published online 27 October 2014
Regulation of primordial follicle recruitment by cross-talk between the Notch and phosphatase and tensin homologue (PTEN)/AKT pathways 
Lin-Qing Wang, Jing-Cai Liu, Chun-Lei Chen, Shun-Feng Cheng, Xiao-Feng Sun, Yong Zhao, Shen Yin, Zhu-Mei Hou, Bo Pan, Cheng Ding, Wei Shen and Xi-Feng Zhang

The growth of oocytes and the development of follicles require certain pathways involved in cell proliferation and survival. However, the mechanism of primordial follicle recruitment remains largely unknown. Inhibition of the Notch signalling pathway suppressed AKT phosphorylation and the proliferation of granulosa cells. Recruitment of primordial follicles was regulated by an interaction between the Notch and PI3K/AKT signalling pathways.

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    | Supplementary Material (167 KB)
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Published online 13 October 2014
Leptin and leptin receptor are detectable in equine spermatozoa but are not involved in in vitro fertilisation 
Anna Lange-Consiglio, Bruna Corradetti, Claudia Perrini, Davide Bizzaro and Fausto Cremonesi

Leptin had effect on capacitation and acrosome reaction in human and swine spermatozoa. In this study, for the first time, mRNAs for leptin and its receptors, and their corresponding proteins were detected in equine spermatozoa. When leptin was used in our in vitro fertilisation model, results show that leptin is involved in capacitation and hyperactivation of equine spermatozoa but not in spermatozoa penetration.

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Published online 09 October 2014
Determinants of valid measurements of global changes in 5ʹ-methylcytosine and 5ʹ-hydroxymethylcytosine by immunolocalisation in the early embryo 
J. Salvaing, Y. Li, N. Beaujean and C. O'Neill

After fertilisation, the embryonic genome must undergo heavy DNA methylation and hydroxymethylation changes that are critical for its development to term. Herein, we focus on the main method used to study these epigenetic modifications in early embryos. Our work emphasises the pitfalls of this method and shows that contradictions observed in the literature can be explained to a considerable extent by its incorrect use.

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Published online 09 October 2014
Long term-cultured and cryopreserved primordial germ cells from various chicken breeds retain high proliferative potential and gonadal colonisation competency 
Céline Tonus, Karine Cloquette, Fabien Ectors, Joëlle Piret, Laurent Gillet, Nadine Antoine, Daniel Desmecht, Alain Vanderplasschen, Olivier Waroux and Luc Grobet

The ability to derive, expand and cultivate avian primordial germ cells (PGCs) without altering their biology is a prerequisite for germplasm-based avian genetic preservation and engineering purposes. We describe original and robust methods that allow long-term expansion and cryobanking of primary cultures of PGCs without obvious effects on their biological properties or their ability to colonise the genital ridges. This provides an alternative access to applications in avian biotechnology and preservation of genetic resources.

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Published online 30 September 2014
Changes in myometrial expression of progesterone receptor membrane components 1 and 2 are associated with human parturition at term 
Ray Wang, Penelope M. Sheehan and Shaun P. Brennecke

The progesterone receptor membrane components 1 and 2 (PGRMC1, PGRMC2) may be a part of the mechanism by means of which women come into labour despite continued high progesterone concentrations. Expression of PGRMC1 mRNA and protein was found to be significantly lower in myometrium from women in labour. This change may be mediated by microRNA species hsa-miR-520e.

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Published online 25 September 2014
Ultrastructure and mitochondrial numbers in pre- and postpubertal pig oocytes 
Hanne Skovsgaard Pedersen, Henrik Callesen, Peter Løvendahl, Fenghua Chen, Jens Randel Nyengaard, Nanett Kvist Nikolaisen, Peter Holm and Poul Hyttel

Prepubertal pig oocytes, used for the production of transgenic pigs as human disease models, have low developmental competence. The aim of this experiment was to describe oocyte ultrastructure and to quantify mitochondria in pre- and postpubertal pig oocytes. The ultrastructure and higher number of mitochondria in large pre- and postpubertal oocytes was compatible with higher developmental competence.

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Published online 24 September 2014
Oncostatin M and leukaemia inhibitory factor trigger signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 pathways but result in heterogeneous cellular responses in trophoblast cells 
Wittaya Chaiwangyen, Stephanie Ospina-Prieto, Diana M. Morales-Prieto, Francisco Lazaro Pereira de Sousa, Jana Pastuschek, Justine S. Fitzgerald, Ekkehard Schleussner and Udo R. Markert

Leukemia inhibitory factor (LIF) and Oncostatin M (OSM) play important roles during implantation and placentation. This study aims to compare responses on LIF and OSM in four different trophoblastic cell lines. In all, LIF and OSM induce STAT3 and ERK1/2 phosphorylation, while only LIF significantly increases trophoblast cell invasion. Thus, both factors have similar but not identical funtions in placentation.

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Published online 19 September 2014
Decreased expression of aquaporin 2 is associated with impaired endometrial receptivity in controlled ovarian stimulation 
Dan Zhang, Gufeng Xu, Runju Zhang, Yimin Zhu, Huijuan Gao, Caiyun Zhou, Jianzhong Sheng and Hefeng Huang

This study revealed the role of auquaporin 2 (AQP2, a water channel) in impaired embryo implantation following ovarian stimulation. It was found that AQP2 was decreased in endometria of ovarian stimulated women and suppressing AQP2 led to impaired implantation. The study indicated the importance of water transportation in embryo implantation and suggested a new way to improve outcome of assisted conception.

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Published online 12 September 2014
Decreased melatonin levels and increased levels of advanced oxidation protein products in the seminal plasma are related to male infertility 
Ewa Maria Kratz, Agnieszka Piwowar, Michal Zeman, Katarína Stebelová and Theresia Thalhammer

Melatonin is a wide-spectrum antioxidant; therefore, the aim of this study was to correlate advanced oxidation protein products (AOPPs) and the total antioxidant capacity (TAC) with melatonin levels in the seminal plasma of men with azoospermia, theratozoospermia and fertile controls. Decreased levels of melatonin and AOPP altered the oxidative–antioxidative balance in the ejaculate, thereby reducing fertility. These parameters may serve as additional markers of semen quality and male reproductive potential.

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Published online 11 September 2014
Discovery of human posterior head 20 (hPH20) and homo sapiens sperm acrosome associated 1 (hSPACA1) immunocontraceptive epitopes and their effects on fertility in male and female mice 
Xuemei Chen, Xiaodong Liu, Xiuhua Ren, Xuewu Li, Li Wang and Weidong Zang

Low antibody titres and the irreversible suppression of spermatogenesis are current hurdles in the contraceptive vaccine field. The purpose of immunocontraception research is to obtain prominent and reversible contraceptive effect through vaccines administered to both males and females. These peptides could be used in the specific diagnosis and treatment of male and female infertility as well as contraceptive vaccine development.

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    | Supplementary Material (277 KB)
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Published online 11 September 2014
Why we should not select the faster embryo: lessons from mice and cattle 
Alfonso Gutierrez-Adan, Carlee R. White, Ann Van Soom and Mellissa R. W. Mann

Many studies have shown that in vitro culture can negatively impact preimplantation development, which necessitates some selection criteria to identify the best-suited embryos for transfer. With emerging data in bovine and mouse supporting increased developmental competency in embryos with moderate rates of development, it is time to question whether the fastest developing embryos are the best embryos for transfer in the human clinic. In this review we explore evidence surrounding the question of developmental timing as it relates to bovine embryo quality, mouse embryo quality and genomic imprint maintenance, and embryo sex.

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Published online 11 September 2014
The effects of permeating cryoprotectants on intracellular free-calcium concentrations and developmental potential of in vitro-matured feline oocytes 
Jason R. Herrick, Chunmin Wang and Zoltan Machaty

Changes in intracellular concentrations of Ca2+, hardening of the zona pellucida, parthenogenetic activation and embryonic development following IVF were evaluated in feline oocytes after exposure to three common cryoprotectants.  A combination of DMSO and propanediol had minimal effects on the viability of feline oocytes and may be useful for the development of more efficient protocols for vitrification of feline oocytes.

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Published online 11 September 2014
Effects of whole-body heat on male germ cell development and sperm motility in the laboratory mouse 
H. Wechalekar, B. P. Setchell, K. R. Pilkington, C. Leigh, W. G. Breed and E. Peirce

Putative effects of whole-body heat exposure include changes in testicular cells and spermatozoa. This study compared laboratory mice exposed to whole-body heat to control mice. The results show reduced testis weights and damage to developing and formed spermatozoa. These changes can cause male infertility. The results from this study could be extrapolated to humans exposed to high temperatures in their occupations.

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Published online 09 September 2014
Epigenetic processes in the male germline 
Alan M. O'Doherty and Paul A. McGettigan

Epigenetic modification of the mammalian genome is indispensable for successful development.  Many of these modifications, such as DNA methylation, are established and removed during gametogenesis, following fertilisation and during the early stages of embryogenesis.  These epigenetic modifications occur at different times during meiosis, depending on whether they occur during spermatogenesis or oogenesis.  Similarly, post-fertilisation modification dynamics occur differently depending on the parental origin of the genomic material.  This review will focus on these processes in the male germline.

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Published online 05 September 2014
Role of Wnt signalling in early pregnancy 
Sarmah B. Nayeem, Frank Arfuso, Arun Dharmarajan and Jeffrey A. Keelan

The Wnt family of secreted ligands plays a major role in regulating a wide range of biological processes, including reproduction. This review summarises our present knowledge of the expression, regulation and function of Wnt signalling pathways in early pregnancy and placentation. Knowledge in this area has applications in the diagnosis, prevention and treatment of infertility and gestational diseases.

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Published online 02 September 2014
Prolactin modulates luteal regression from the coeliac ganglion via the superior ovarian nerve in the late-pregnant rat 
Sandra S. Vallcaneras, Magalí de la Vega, Silvia M. Delgado, Alicia Motta, Carlos Telleria, Ana M. Rastrilla and Marilina Casais

This work provides evidence of the functional interaction between the endocrine and peripheral neural systems. The aim was to study the effects of prolactin added to the coeliac ganglion, via the superior ovarian nerve, on ovarian physiology at the end of rat pregnancy. Prolactin acts at the coeliac ganglion and sends information to the ovary by a neural pathway, modifying parameters of functional and structural luteal regression.

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Published online 28 August 2014
Stress preconditioning of semen before cryopreservation improves fertility and increases the number of offspring born: a prospective randomised study using a porcine model 
A. Horváth, O. Szenci, K. Nagy, L. Végh and Cs. Pribenszky

Standard cryopreservation protocols and modified AI techniques are required to achieve acceptable fertility results with frozen–thawed boar semen. Using simplified procedures, but applying a sublethal hydrostatic pressure to the semen at room temperature before cryopreservation, yielded higher motility and fertility results. The AI dosage still needs to be optimised and, because no direct correlation was found between motility and fertility on an individual basis, the motility results cannot be used to predict potential fertility.

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Published online 28 August 2014
Seminal plasma affects sperm sex sorting in boars 
Diego V. Alkmin, Inmaculada Parrilla, Tatiana Tarantini, David del Olmo, Juan M. Vazquez, Emilio A. Martinez and Jordi Roca

Sex-sorted spermatozoa can play a crucial role in enhancing genetic improvement and production efficiency in swine industry. The presence of a high proportion of seminal plasma in semen samples negatively influences sperm sex-sorting efficiency. Removing the seminal plasma before sorting can reverse the consistent poor sperm sortability exhibited by the ejaculates of some boars.

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Published online 28 August 2014
Human extravillous trophoblast invasion: intrinsic and extrinsic regulation 
E. Menkhorst, A. Winship, M. Van Sinderen and E. Dimitriadis

Invasion of specialised fetal cells, the extravillous trophoblast, into the uterine decidua is critical for the establishment of pregnancy. Overall, there exists an exquisite balance between trophoblast over- and under-invasion. This review summarises what is known about the intrinsic invasive potential of the extravillous trophoblast and the role of maternal cells in regulating invasion. A greater understanding of the mechanisms will lead to therapeutics to treat at-risk pregnancies.

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Published online 26 August 2014
Signalling pathways involved in the synergistic effects of human growth differentiation factor 9 and bone morphogenetic protein 15 
Karen L. Reader, David G. Mottershead, Georgia A. Martin, Robert B. Gilchrist, Derek A. Heath, Kenneth P. McNatty and Jennifer L. Juengel

The oocyte-derived growth factors, bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), play important roles in fertility, but their mechanism of action differs between species. Our aim was to determine the signalling pathways used by human BMP15 and GDF9; we found that they act synergistically and the combinations of pathways used are different from those observed in other species. Understanding the mechanisms of function of these human growth factors may help tailor treatments to improve fertility.

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Published online 25 August 2014
Interleukin 11: similar or opposite roles in female reproduction and reproductive cancer? 
Amy Winship, Ellen Menkhorst, Michelle Van Sinderen and Evdokia Dimitriadis

A signalling protein important in pregnancy can also promote uterine tumour growth in women. High levels of interleukin (IL)-11 protein are found in the placenta of pregnant women who have developed the potentially life-threatening condition of pre-eclampsia. The same protein is also highly expressed in tumours that develop in the endometrium, the inner lining of the uterus. IL-11 may be a new potential therapeutic target for cancer and the pregnancy complication pre-eclampsia.

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Published online 22 August 2014
Oxygen modulates human embryonic stem cell metabolism in the absence of changes in self-renewal 
Alexandra J. Harvey, Joy Rathjen, Lijia Jackie Yu and David K. Gardner

In this study we characterise the ability of environmental oxygen to regulate human embryonic stem cells (hESC). We show that the oxygen concentration influences metabolite use, metabolic gene expression, cell death and free radical production in hESC, without affecting the expression of characteristic hESC markers. Our findings suggest that hESC benefit from culture in a reduced oxygen environment that mimics the physiological paradigm.

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Published online 22 August 2014
Role of linker histone H1c during the reprogramming of Chinese swamp buffalo (Bubalus Bubalis) embryos produced by somatic cell nuclear transfer 
Gao-Bo Huang, Li Quan, Yong-Lian Zeng, Jian Yang, Ke-Huan Lu and Sheng-Sheng Lu

The movement of numerous proteins between the cytoplasm and nucleus is essential to remodelling. In this study, the exchange of histone between the cytoplasm of the oocyte (H1foo) and the transferred nucleus (H1c) is shown to be essential to somatic cell nuclear transfer in buffalo.

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    | Supplementary Material (237 KB)
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Published online 20 August 2014
Variation in seminal plasma alters the ability of ram spermatozoa to survive cryopreservation 
J. P. Rickard, R. E. Schmidt, J. W. Maddison, R. Bathgate, G. W. Lynch, X. Druart and S. P. de Graaf

The effect of seminal plasma on sperm function is yet to be fully explained. This study demonstrates that survival of spermatozoa during cryopreservation is modified by the source of seminal plasma (high or low freezing resilient male) to which they are exposed prior to freezing. These results demonstrate that seminal plasma varies in composition and is the first step in identifying its positive and negative components.

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    | Supplementary Material (24 KB)
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Published online 13 August 2014
Analysis of STAT1 expression and biological activity reveals interferon-tau-dependent STAT1-regulated SOCS genes in the bovine endometrium 
A. Vitorino Carvalho, C. Eozenou, G. D. Healey, N. Forde, P. Reinaud, M. Chebrout, L. Gall, N. Rodde, A. Lesage Padilla, C. Giraud Delville, M. Leveugle, C. Richard, I. M. Sheldon, P. Lonergan, G. Jolivet and O. Sandra

Implantation is a critical step of mammalian pregnancy whose progression depends on the quality of embryo and female genital tract. Using cattle as an animal model, gene expression, regulation and cell localization of STAT1 transcription factor were investigated in the endometrium during the oestrous cycle and early pregnancy; direct STAT1 target genes were identified by chromatin immunoprecipitation run with primary cultures of bovine endometrial cells. Altogether our data provide new insights on STAT1 biological functions in the endometrium during pregnancy maternal recognition and implantation.

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    | Supplementary Material (325 KB)
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Published online 13 August 2014
Low physiological levels of prostaglandins E2 and F improve human sperm functions 
Mariana Rios, Daniela V. Carreño, Carolina Oses, Nelson Barrera, Bredford Kerr and Manuel Villalón

The recent discovery that prostaglandins are present in the fluid of the female genital tract and increase the motility of human sperm in vitro could lead to a possible treatment to improve male infertility. The findings of this study that incubation of human spermatozoa with low physiological levels of PGE2 or PGF increases sperm functions could improve conditions for assisted reproduction protocols and help infertile couples seeking medical help, which, according to global estimates, could be as many as 50–80 million people.

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Published online 13 August 2014
The requirement for protein kinase C delta (PRKCD) during preimplantation bovine embryo development 
Qi-En Yang, Manabu Ozawa, Kun Zhang, Sally E. Johnson and Alan D. Ealy

Embryonic loss is one of the major contributors to infertility in humans and domestic animals. This work determined that protein kinase C delta, an intracellular signalling molecule, is involved in several important developmental processes during early embryonic development in cattle. These outcomes implicate this molecule as a principal player in establishing the developmental potential of bovine embryos.

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    | Supplementary Material (520 KB)
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Published online 11 August 2014
A new technique for analysis of human sperm morphology in unstained cells from raw semen 
Carles Soler, Almudena García-Molina, María Sancho, Jesús Contell, Manuel Núñez and Trevor G. Cooper

We describe the first technique that makes it possible to analyse the real size and shape of sperm cells. The development and application of this technique must lead to a new classification of human sperm morphology. This new approach will provide a revolutionary tool in the definition of human male fertility.

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Published online 11 August 2014
Yolk size and ovulation order determine fertility within double-yolked duck (Anas platyrhynchos domesticus) eggs 
Attila Salamon and John P. Kent

In double-yolked duck eggs, the first yolk ovulated achieved a higher level of fertilisation (68.82%) than the second yolk (34.98%), demonstrating a primacy effect. Eggs with larger yolks achieved a higher level of fertilisation, demonstrating an optimal yolk size effect. Both primacy and optimal size have a positive effect on the successful fertilisation of eggs in species with internal fertilisation.

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Published online 01 August 2014
Effects of maternal undernutrition during late gestation and/or lactation on colostrum synthesis and immunological parameters in the offspring 
S. Chadio, A. Katsafadou, B. Kotsampasi, G. Michailidis, K. C. Mountzouris, D. Kalogiannis and V. Christodoulou

Colostrum, apart from its crucial role for neonate survival, serves as a conduit for the transmission of signalling molecules from the mother to offspring. Maternal nutrition during late gestation affects colostrum composition and immune factor content, which through immunomodulatory properties, alters immune system function in the offspring. Events during fetal or neonatal life may have programming effects on immune function in later life.

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Published online 28 July 2014
Expression of group III metabotropic glutamate receptors in the reproductive system of male mice 
Marcin Marciniak, Barbara Chruscicka, Tomasz Lech, Grzegorz Burnat and Andrzej Pilc

Although glutamate signalling is traditionally investigated in the context of the central nervous system, it could also play a role in non-neuronal tissues. Herein, we investigated the expression of group III metabotropic glutamate (mGlu) receptors in the reproductive system of male mice. Based on findings for the mGlu7 receptor, we postulate that constitutively expressed mGlu receptors in the male reproductive system may influence male fertility.

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Published online 24 July 2014
Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Müllerian hormone profiles 
G. Gamarra, C. Ponsart, S. Lacaze, B. Le Guienne, P. Humblot, M.-C. Deloche, D. Monniaux and A. A. Ponter

Ovum pick-up and in vitro embryo production is a technique that can be used in high genetic merit dairy heifers to produce large numbers of high quality embryos. Serum anti-Müllerian hormone (AMH) was positively correlated with the numbers and quality of in vitro produced embryos after superovulation. A simple dietary supplement, propylene glycol, which increased circulating insulin, glucose and IGF1, was able to further increase oocyte quality and embryo production in animals with high AMH levels.

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Published online 24 July 2014
Variations in epithelial Na+ transport and epithelial sodium channel localisation in the vaginal cul-de-sac of the brushtail possum, Trichosurus vulpecula, during the oestrous cycle 
T.-A. Alsop, B. J. McLeod and A. G. Butt

The vaginal cul-de-sac, which is unique to the reproductive tract of marsupials, is thought to play an important role in reproduction, as the volume of fluid in the cul-de-sac increases up to ovulation and diminishes thereafter. Generally, changes in fluid volume in the reproductive tract are a result of changes in the transport properties of the epithelial lining. Our findings describe an epithelial transport mechanism responsible for absorption of fluid from the cul-de-sac following ovulation.

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Published online 22 July 2014
N-Glycans in Xenopus laevis testis characterised by lectin histochemistry 
Galder Valbuena, Juan Francisco Madrid, María Martínez de Ubago, Laura Gómez-Santos, Edurne Alonso, Lucio Díaz-Flores and Francisco J. Sáez

N-glycans (oligosaccharide chains linked to proteins by the amino group of an asparagine) essentials for mammalian spermatogenesis have been identified. Here, for the first time the presence of N-glycans in the testis of the amphibian Xenopus laevis has been analysed by lectin histochemistry. The functional role of these glycans could be related to the role of N-glycans involved in mammalian spermatogenesis.

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Published online 18 July 2014
Effect of combined exogenous progesterone with luteotrophic support via equine chorionic gonadotrophin (eCG) on corpus luteum development, circulating progesterone concentrations and embryo development in cattle 
L. O'Hara, N. Forde, P. Duffy, F. Randi, A. K. Kelly, A. Valenza, P. Rodriguez and P. Lonergan

Low circulating progesterone in the week following ovulation has been implicated in poor fertility in cattle. The aim of this study was to examine the effect of exogenous progesterone in combination with a luteotrophic agent on corpus luteum survival and conceptus development in cross-bred beef heifers. Results may provide the basis for a strategy to elevate progesterone and improve fertility.

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Published online 18 July 2014
Superovulation alters embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) gene expression in mouse oocytes and early embryos 
Saffet Ozturk, Aylin Yaba-Ucar, Berna Sozen, Derya Mutlu and Necdet Demir

Superovulation adversely influences oocyte maturation and early embryo development, but how superovulation results in these disturbances has not been clearly addressed yet. In this study, we aimed to determine the effect of superovulation on expression of embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) genes. We found that superovulation significantly alters Epab and Pabpc1 gene expression in mouse oocytes and early embryos.

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Published online 18 July 2014
In vitro interaction between resistin and peroxisome proliferator-activated receptor γ in porcine ovarian follicles 
Agnieszka Rak-Mardy?a and Eliza Drwal

Resistin is a small cysteine-rich protein, produced by adipose tissue and involved in insulin resistance, type 2 diabetes and reproduction. The findings of this study suggest that there is local cooperation exists resistin and PPARγ expression in the porcine ovary; resistin significantly increases PPARγ expression, whereas PPARγ decreases resistin expression. PPARγ is a new key regulator of resistin expression and function.

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Published online 18 July 2014
Expression patterns of mitochondrial OXPHOS components, mitofusin 1 and dynamin-related protein 1 are associated with human embryo fragmentation 
Vesna Otasevic, Lela Surlan, Milica Vucetic, Ivan Tulic, Biljana Buzadzic, Ana Stancic, Aleksandra Jankovic, Ksenija Velickovic, Igor Golic, Milica Markelic, Aleksandra Korac and Bato Korac

Improving the quality of preimplantation embryos is imperative in human in vitro fertilisation since the birth of the first baby in 1978, and high fragmentation is one of the limiting factors. This study links the phenomenon of human embryo fragmentation with energy homeostasis, mitochondrial dynamics and oxidative phosphorylation, suggesting altered transcriptional profiles of mitochondria-encoded components. A better understanding of the molecular mechanisms underlying these processes may help improve the energy capacity of the early embryo.

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Published online 17 July 2014
Mitochondrial activity during pre-maturational culture in in vitro-grown bovine oocytes is related to maturational and developmental competences 
Weiping Huang, Sung-Sik Kang, Katsuhisa Nagai, Yojiro Yanagawa, Yoshiyuki Takahashi and Masashi Nagano

Clarifying the mechanism for acquisition of developmental competence in bovine oocytes is important for improving the in vitro production system. Relationships between developmental competence and mitochondrial activity of in vitro-grown oocytes during meiotic arrest immediately before IVM culture were investigated. Mitochondrial activity at 10 h after starting meiotic arrest was highest and those oocytes showed comparable developmental competence with in vivo-grown oocytes.

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Published online 16 July 2014
Efficient generation of sFat-1 transgenic rabbits rich in n-3 polyunsaturated fatty acids by intracytoplasmic sperm injection 
Shun Zhang, Fenghua Lu, Qingyou Liu, Yubing Liu, Xiaomei Guan, Yingming Wei, Shijian Tan and Deshun Shi

N-3 PUFAs are beneficial to human health, and increasing the amount of n-3 PUFAs in mammals by introducing sFat-1 gene will improve the animal meat quality. This study was undertaken to produce sFat-1 transgenic rabbit by ICSI, and found that sFat-1 transgenic rabbits exhibited an ~10-fold increase in n-3 PUFAs in muscles. It will be possible to produce a new transgenic rabbit breed with high n-3 PUFAs.

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Published online 15 July 2014
Low-density lipoprotein receptor affects the fertility of female mice 
Tao Guo, Liang Zhang, Dong Cheng, Tao Liu, Liguo An, Wei-Ping Li and Cong Zhang

Low density lipoprotein receptor (LDLR) is responsible for the uptake of cholesterol-carrying lipoprotein particles into cells. The Ldlr deficient mice developed severe dyslipidemia and had significantly decreased female fertility. This study provides additional evidence for the role of Ldlr in regulation of lipid metabolism as well as female reproductive health during postnatal life.

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Published online 15 July 2014
Low level of basal testosterone: a significant risk factor for poor oocyte yield after ovulation induction 
Jing Guo, Qingxue Zhang, Yu Li, Wenjun Wang and Dongzi Yang

This study supplies a new predictor for poor ovarian response (POR) in the in vitro fertilisation field. We investigated the association of low basal androgen levels and IVF outcomes in a large Chinese population, and found low basal testosterone (<0.88 nmol L–1) was an independent risk factor for POR. This finding may help to identify young patients at high risk of POR.

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Published online 15 July 2014
Oocytes recovered after ovarian tissue slow freezing have impaired H2AX phosphorylation and functional competence 
Sam Sudhakaran, Shubhashree Uppangala, Sujith Raj Salian, Sachin D. Honguntikar, Ramya Nair, Guruprasad Kalthur and Satish Kumar Adiga

Ovarian tissue cryopreservation by slow freezing and vitrification are techniques available to cancer-affected women to preserve their fertility before cancer therapy. Comparison of these two techniques in mouse ovarian tissue has demonstrated the adverse effects of ovarian tissue slow freezing but not vitrification on oocyte DNA quality and functional ability. Hence, vitrification may be preferred over slow freezing for ovarian tissue cryopreservation.

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Published online 11 July 2014
Analysis and characterisation of bovine oocyte and embryo biomarkers by matrix-assisted desorption ionisation mass spectrometry imaging 
Roseli F. Gonçalves, Mónica S. Ferreira, Diogo N. de Oliveira, Rafael Canevarolo, Marcos A. Achilles, Daniela L. D'Ercole, Peter E. Bols, Jose A. Visintin, Gary J. Killian and Rodrigo R. Catharino

Lipid pathways play important biological roles in mammalian embryology. We document a new strategy to analyse bovine oocytes and embryos, and provide a chemical image using MALDI-MSI technology. Results obtained in this work demonstrates that a combination of MF, MSI features and chemometric analysis can be applied to discriminate cell stages, characterising specific biomarkers and relating them to developmental pathways.

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    | Supplementary Material (126 KB)
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Published online 09 July 2014
Differential gene-expression profiles from canine cumulus cells of ovulated versus in vitro-matured oocytes 
Su-Jin Cho, Kyeong-Lim Lee, Yu-Gon Kim, Dong-Hoon Kim, Jae-Gyu Yoo, Byoung-Chul Yang, Jin-Ki Park and Il-Keun Kong

Canine oocytes are ovulated at the GV/GVBD stage and rely on CCs to mature to MII stage. Consequently, the role of CCs is very likely to be more important than in other species. The objective was to identify the relationship between oocytes and cumulus cells during oocyte maturation. The results demonstrated that cumulus cells play key roles in supporting oocyte maturation to the MII stage.

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    | Supplementary Material (41 KB)
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Published online 08 July 2014
Semen collection, ejaculate characteristics and in vitro manipulation of spermatozoa from six species of captive flying-fox (Pteropus spp.) 
D. F. Melville, E. G. Crichton and S. D. Johnston

Almost two thirds of the 65 documented Pteropus species (large bats called flying-foxes) are considered threatened with extinction. Our research aimed to provide foundational information from which to develop a method of semen collection, extension and preservation for use in male fertility assessments and artificial insemination. The successful collection of >400 semen samples from six Pteropus species, four which had never been characterised before, adds to the general knowledge of male reproductive biology in these species and advances the likelihood that artificial reproductive technologies may one day be used to assist conservation efforts.

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Published online 26 June 2014
Impacts of and interactions between environmental stress and epigenetic programming during early embryo development 
Michael J. Bertoldo, Yann Locatelli, Christopher O'Neill and Pascal Mermillod

Embryos experience stress when they are grown in the laboratory, and this is believed to be detrimental to the embryo later in life. This article reviews the effects of in vitro conditions on the embryo and the mechanisms involved. By having a greater understanding of the mechanisms, we can potentially prevent stress during embryo growth in vitro.

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Published online 26 June 2014
Effects of varying doses of Β-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos) 
C. C. Stuart, J. L. Vaughan, C. M. Kershaw-Young, J. Wilkinson, R. Bathgate and S. P. de Graaf

Alpaca seminal plasma contains the protein β-nerve growth factor (β-NGF), which induces ovulation in females. We examined the effects of three doses of β-NGF and a gonadotrophin-releasing hormone (GnRH) agonist on ovulation rate and interval, corpus luteum diameter and plasma progesterone when administered to female alpacas and found that only ovulation rate varied. This suggests that there is no advantage to using β-NGF over more readily available and cheaper GnRH agonists to induce ovulation in alpacas.

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Published online 26 June 2014
Antibiotics and oxygen availability affect the short-term storage of spermatozoa from the critically endangered booroolong frog, Litoria booroolongensis 
Aimee J. Silla, Leesa M. Keogh and Phillip G. Byrne

Sperm storage technologies aim to extend sperm longevity and increase the time available to achieve artificial fertilisation. Overall, results from this study on the booroolong frog indicate that increased oxygen availability is beneficial to sperm longevity, but that antibiotics are detrimental. The optimal storage protocols identified will allow spermatozoa from spatially separated breeding stock to be transferred among captive facilities, a tool that will assist the genetic management of this critically endangered species.

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Published online 23 June 2014
Suboptimal maternal nutrition during early-to-mid gestation in the sheep enhances pericardial adiposity in the near-term fetus 
Shalini Ojha, Michael E. Symonds and Helen Budge

Changes in maternal diet during pregnancy can have long-lasting influences on the offspring’s health. In sheep, suboptimal nutrition during early-to-mid gestation results in enhanced fat deposition around the fetal heart. This is accompanied by increased abundance of genes that promote adipocyte synthesis. These adaptations may promote survival at birth but contribute to obesity in later life.

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Published online 19 June 2014
Effect of superstimulation protocols on nuclear maturation and distribution of lipid droplets in bovine oocytes 
D. Dadarwal, M. Honparkhe, F. C. F. Dias, T. Alce, C. Lessard and J. Singh

Lipids are the main energy source for oocytes. Based on 3D-microscopic evaluation, a mature bovine oocyte contained 22 picolitres of lipids distributed as 3000 droplets occupying 3.3% of the cell volume. Extending the FSH treatment from 4 to 7 days yielded a higher proportion of mature oocytes with no effect on lipids. FSH starvation for 84 h resulted in failure of meiotic resumption and accumulation of large lipid droplets, indicating degenerative changes.

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Published online 19 June 2014
Free-radical production after post-thaw incubation of ram spermatozoa is related to decreased in vivo fertility 
Enrique Del Olmo, Alfonso Bisbal, Olga García-Álvarez, Alejandro Maroto-Morales, Manuel Ramón, Pilar Jiménez-Rabadán, Luis Anel-López, Ana J. Soler, J. Julián Garde and María R. Fernández-Santos

This paper shows a direct relation between fertility and ROS production in sperm samples. The Fluorescein (CM-H2DCFDA) test combined with the SOF incubation could be considered a standard method for improving sperm-quality assessment. Ultimately, this technique could be a possible solution to the problem of fertility prediction in the laboratory.

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    | Supplementary Material (70 KB)
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Published online 16 June 2014
Insulin regulates primordial-follicle assembly in vitro by affecting germ-cell apoptosis and elevating oestrogen 
Xin-Lei Feng, Yuan-Chao Sun, Min Zhang, Shun-Feng Cheng, Yan-Ni Feng, Jing-Cai Liu, Hong-Hui Wang, Lan Li, Guo-Qing Qin and Wei Shen

Insulin plays an important role in the regulation of ovarian function; however, the specific molecular mechanism of its function remains largely unknown. This study found that insulin promoted folliculogenesis by facilitating germ-cell apoptosis within the cysts and upregulating oestrogen levels. This work established an effective model to study the function of insulin in folliculogenesis.

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    | Supplementary Material (285 KB)
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Published online 12 June 2014
Transgenerational inheritance of ovarian development deficiency induced by maternal diethylhexyl phthalate exposure 
Xi-Feng Zhang, Teng Zhang, Zhe Han, Jing-Cai Liu, Yu-Ping Liu, Jun-Yu Ma, Lan Li and Wei Shen

Diethylhexyl phthalate (DEHP) disrupts the physiological functions of endogenous hormones and induces abnormal development of mammals. However, the effects of DEHP exposure on folliculogenesis remain largely unknown. DEHP exposure significantly delayed the first meiotic progression of female fetal germ cells and accelerated follicle recruitment. DEHP exposure induced ovarian development deficiency, which was transgenerational in mice.

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    | Supplementary Material (581 KB)
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Published online 03 June 2014
Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay 
K. Pollock, J. Gosálvez, F. Arroyo, C. López-Fernández, M. Guille, A. Noble and S. D. Johnston

The integrity of sperm DNA is recognised as an important parameter of semen quality, but there are no published reports of this procedure for any amphibian. Validated as the first amphibian protocol for the assessment of sperm DNA fragmentation, the sperm chromatin dispersion test revealed three nuclear morphotypes representing varying degrees of DNA damage. This protocol has the potential to benefit assisted breeding programs using assisted reproductive technologies to support amphibian conservation efforts.

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Published online 29 May 2014
Open pulled straw vitrification and slow freezing of sheep IVF embryos using different cryoprotectants 
M. H. Bhat, V. Sharma, F. A. Khan, N. A. Naykoo, S. H. Yaqoob, G. Vajta, H. M. Khan, M. R. Fazili, N. A. Ganai and R. A. Shah

Embryo cryopreservation has huge potential to ensure the storage and worldwide distribution of elite germplasm. The aim of this study was to establish a standard procedure using specific cryoprotectant concentrations to avoid loss during cryopreservation procedures. Vitrification using 40% ethylene glycol resulted in superior embryo survival in sheep embryos from abattoir-derived oocytes. These results provide an easier option for embryo cryopreservation without compromising embryo survival.

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Published online 22 May 2014
Protein deubiquitination during oocyte maturation influences sperm function during fertilisation, antipolyspermy defense and embryo development 
Young-Joo Yi, Miriam Sutovsky, Won-Hee Song and Peter Sutovsky

Tagging with multi-ubiquitin chains targets substrate proteins for recycling by the 26S proteasome, during which deubiquitinating enzymes (DUBs) remove multi-ubiquitin chains from the substrate to regenerate mono-ubiquitin. In this study, DUB inhibitors impaired porcine oocyte maturation, fertilisation and pre-embryo development by altering the turnover of oocyte proteins such as major vault protein. Thus, DUBs may promote the acquisition of oocyte developmental competence.

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Published online 28 April 2014
Distribution of cortical granules and meiotic maturation of canine oocytes in bi-phasic systems 
Maricy Apparicio, Giuliano Q. Mostachio, Tathiana F. Motheo, Aracelle E. Alves, Luciana Padilha, Eliandra A. Pires-Butler, Paula A. P. Savi, Ricardo A. R. Uscategui, Gaia C. Luvoni and Wilter R. R. Vicente

Studies about canine reproduction have increased over the years once this species can be used as a model for their endangered counterparts. This work aimed to reproduce in laboratory a similar hormonal condition in order to improve the results that are still far from satisfactory. Our results reveal that mimicking the hormonal changes is beneficial for oocyte maturation and so it is a more reliable approach to overcome the challenges of in vitro techniques.

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Published online 23 April 2014
Src family kinases are involved in the meiotic maturation of porcine oocytes 
Kateřina Kheilová, Jaroslav Petr, Tereza Žalmanová, Veronika Kučerová-Chrpová and Dalibor Řehák

Src family kinases are involved in the meiotic maturation of mammalian oocytes, but the information about their role in regulation of meiotic maturation has been mainly obtained from mouse oocytes. The present study proved that the Src family kinases play an active role in the meiotic maturation of porcine oocytes at least partly through MPF and MAPK activity regulation. Considering that data from mouse oocytes need not be validated in other species, our findings contribute to the understanding of the regulation of oocyte maturation.

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Published online 17 April 2014
Addition of l-ascorbic acid to culture and vitrification media of IVF porcine blastocysts improves survival and reduces HSPA1A levels of vitrified embryos 
Miriam Castillo-Martín, Marc Yeste, Albert Soler, Roser Morató and Sergi Bonet

The presence of non-enzymatic antioxidants in the environment surrounding the embryo reduces oxidative stress. Thus, the present study sought to determine how the addition of l-ascorbic acid during culture and vitrification and warming affects quality and gene expression of in vitro produced porcine blastocysts. Results obtained indicate that l-ascorbic acid supplementation increases embryo survival following cryopreservation via decreasing HSPA1A levels.

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Published online 10 April 2014
Substitution of starch for palm oil during gestation: impact on offspring survival and hepatic gene expression in the pig 
K. L. Almond, H. P. Fainberg, M. A. Lomax, P. Bikker, M. E. Symonds and A. Mostyn

Various dietary strategies to reduce piglet mortality have been tested, with increased fat in the maternal diet thought to improve energy partitioning to the fetus, subsequently improving neonatal outcome. However, we found that a maternal diet containing palm oil in place of starch led to reduced maternal glucose tolerance, increased neonatal mortality and altered gene expression in the liver at 1 week of age. Despite this, offspring from mothers fed the fat-substituted diet displayed no changes in postnatal growth or hepatic gene expression at 6 months of age.

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Published online 10 April 2014
In utero exposure to the oestrogen mimic diethylstilbestrol disrupts gonadal development in a viviparous reptile 
Laura M. Parsley, Erik Wapstra and Susan M. Jones

Studies of endocrine-disrupting chemicals (EDCs) in egg-laying reptiles have significantly advanced knowledge in this field; however, 30 % of reptiles are live-bearing. We examined the effects of feminising EDCs by exposing female Niveoscincus metallicus to the synthetic oestrogen, diethylstilbestrol, and found that gonadal development of male and female neonates from exposed mothers was impaired at birth. Maternal exposure to feminising EDCs may have profound consequences on reproductive fitness.

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Published online 10 April 2014
Antioxidant treatment during manipulation procedures prevents mitochondrial and DNA damage and enhances nuclear reprogramming of bovine somatic cell nuclear transfer embryos 
Hyo-Kyung Bae, In-Sun Hwang, Ji-Ye Kim, Sung-Young Lee, Choon-Keun Park, Boo-Keun Yang and Hee-Tae Cheong

The efficiency of animal cloning by somatic cell nuclear transfer (SCNT) is low because of incomplete nuclear reprogramming. We tried to prevent the mitochondrial and DNA damage caused by mechanical stress-associated reactive oxygen species during manipulation procedures of SCNT using antioxidants. Antioxidant treatment during manipulation procedures can efficiently prevent cellular damage and improve nuclear reprogramming.

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Published online 04 April 2014
Prenatal exposure to testosterone masculinises the female gerbil and promotes the development of lesions in the prostate (Skene’s gland) 
Manoel F. Biancardi, Ana P. S. Perez, Cássia Regina Suzuki Caires, Rejane M. Góes, Patrícia S. L. Vilamaior, Fernanda C. A. Santos and Sebastião R. Taboga

This research reports important improvements in prostate biology focusing on the mechanisms underlying the origins of diseases from the female prostate (Skene’s gland). Thus, the aim of this study was to evaluate the interferential effects of exogenous testosterone exposure during the prenatal and pubertal periods on the health of the female prostate. We observed that the exposure to androgenic disruptors may predispose the gland to develop lesions with aging.

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Published online 03 April 2014
Differential expression of Notch component and effector genes during ovarian follicle and corpus luteum development during the oestrous cycle 
D. Murta, M. Batista, E. Silva, A. Trindade, L. Mateus, A. Duarte and L. Lopes-da-Costa

Ovarian dynamics involve extensive cell-to-cell communication, requiring fine regulation of cell signalling. This led us to evaluate the expression patterns of Notch cell-signalling pathway component and effector genes during follicle and corpus luteum development. We found Notch signalling active, and Notch expression spatially and temporally associated with key events in follicle and corpus luteum development. This suggests a relevant regulatory role of Notch signalling in ovarian dynamics.

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Published online 03 April 2014
Effects of vitrification on the expression of pluripotency, apoptotic and stress genes in in vitro-produced porcine blastocysts 
Miriam Castillo-Martín, Marc Yeste, Eva Pericuesta, Roser Morató, Alfonso Gutiérrez-Adán and Sergi Bonet

The demand for the successful preservation of porcine embryos has increased in the past decade. In this study, we evaluated the effects of vitrification and warming, a widely used cryopreservation method, on the expression of six relevant genes in in vitro-produced porcine blastocysts. The results suggest that vitrification and warming affect embryo developmental ability and pluripotency by increasing the number of cells with fragmented DNA, peroxide levels and HSPA1A gene expression.

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Published online 02 April 2014
Seasonal variation in urinary and salivary reproductive hormone levels in Amazonian manatees (Trichechus inunguis) 
Rodrigo S. Amaral, Fernando C. W. Rosas, Vera M. F. da Silva, Laura H. Graham, Priscila Viau, Marcilio Nichi and Claudio A. Oliveira

The Amazonian manatee is a threatened aquatic mammal endemic to the Amazon basin and the species is a seasonal breeder. The aim of this study was to evaluate the effect of the season on the reproductive hormone levels of captive Amazonian manatees. Different from males, all female hormones levels showed seasonal variation. The results strongly suggest the existence of reproductive seasonality in Amazonian manatees, even in captive conditions; however, apparently only females exhibit reproductive quiescence during the non-breeding season.

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Published online 31 March 2014
Effect of pregnancy on endometrial expression of luteolytic pathway components in the mare 
M. de Ruijter-Villani, H. T. A. van Tol and T. A. E. Stout

Preventing pulsatile endometrial PGF release is critical to pregnancy recognition in mares, and downregulation of PTGS2 and OXTR underlies initial avoidance of luteolysis. By Day 21 of pregnancy, however, expression of both PTGS2 and OXTR recovers to levels consistent with luteolysis. Continued luteal maintenance may instead involve uncoupling of PGF generating feedback loops by inhibition of PGF receptor (PTGFR) expression.

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Published online 31 March 2014
Ram spermatozoa migrating through artificial mucus in vitro have reduced mitochondrial membrane potential but retain their viability 
Carmen Martínez-Rodríguez, Mercedes Alvarez, Elena López-Urueña, Susana Gomes-Alves, Luis Anel-López, Cesar A. Chamorro, Luis Anel and Paulino de Paz

The ram sperm population that migrates through a column of cervical mucus surrogates in a penetration test was evaluated. These spermatozoa exhibit an adequate motility to cross the mucus barrier and we think that these cells can give information about the transport through the female genital tract. The viability of migrating spermatozoa was not changed but the mitochondrial status was affected after migration.

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Published online 27 March 2014
Cloning and expression of progesterone receptor isoforms A and B in bovine corpus luteum 
Robert Rekawiecki, Magdalena Karolina Kowalik and Jan Kotwica

Nuclear progesterone receptor has two main isoforms, PGRA and PGRB, which have different functions. PGRB cDNA from cattle was cloned then mRNA and protein levels for both isoforms in corpora lutea during the oestrous cycle and first trimester of pregnancy were determined. The data suggest that the variable expression of these isoforms during the oestrous cycle may depend on the influence of progesterone.

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Published online 25 March 2014
Contribution of boars to reproductive performance and paternity after homospermic and heterospermic artificial insemination 
C. E. R. Ferreira, D. B. Sávio, A. C. Guarise, M. J. Flach, G. D. A. Gastal, A. O. Gonçalves, O. A. Dellagostin, R. V. Alonso, I. Bianchi, C. D. Corcini and T. Lucia

Although it is commonly used in swine farms, heterospermic AI does not allow precise evaluation of boar fertility. The reproductive performance and the paternity contribution of four boars (A, B, C and D) were compared in this study using homospermic and heterospermic AI in 511 sows. Differences among boars were only identified after genotyping the piglets sired through heterospermic AI.

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Published online 25 March 2014
Validation of computer-assisted sperm-motility analysis in the amphibian Silurana tropicalis 
Severine Larroze, Daniel B. Pickford and William V. Holt

Computer-assisted sperm-motility analysis has been scarcely studied or used on amphibians. In this research we validated a standardised method producing reproducible results and compared traditional data analysis with sperm-subpopulation frequencies. We demonstrated that sperm-subpopulation frequency analysis can detect treatment effects on sperm motility.

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Published online 20 March 2014
Mouse ovarian tissue vitrification on copper electron microscope grids versus slow freezing: a comparative ultrastructural study 
Ferda Topal-Celikkan, Sinan Ozkavukcu, Deniz Balci, Sibel Serin-Kilicoglu and Esra Atabenli-Erdemli

Ovarian cryopreservation for fertility preservation is a gold standard procedure in numerous cases, like childhood cancers, although it is still considered experimental. Vitrification is widely used in embryo and oocyte cryopreservation, but its effectiveness for ovarian freezing remains debatable. Slow freezing seems to preserve ovarian content better and therefore the vitrification of tissues needs to be improved by means of cryoprotective agents and carrying devices.

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Published online 17 March 2014
Follistatin is essential for normal postnatal development and function of mouse oviduct and uterus 
S. J. Holdsworth-Carson, R. G. Craythorn, W. R. Winnall, K. Dhaliwal, R. Genovese, C. J. Nowell, P. A. W. Rogers, D. M. de Kretser, M. P. Hedger and J. E. Girling

Follistatin is a protein with key roles in the female reproductive system. We used mutant mouse strains to demonstrate the importance of follistatin in the postnatal development and function of the uterus and oviduct. Our work highlights the need to further examine the role of follistatin in the reproductive tract, as well as its key role in the ovary.

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Published online 11 March 2014
Developmental programming of bone deficits in growth-restricted offspring 
Tania Romano, John D. Wark and Mary E. Wlodek

Offspring born with low birth weight are predisposed to a number of adult diseases, including osteoporosis. Herein, we report that such rat offspring develop reduced bone mineral content and strength at various ages. Importantly these deficits affected males more severely than females and became worse with aging. Interventions to improve bone quality in adult life for individuals born small may reduce the incidence of osteoporosis in the future.

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Published online 04 March 2014
Variation of spermatogenic and Sertoli cell number detected by fine needle aspiration cytology (FNAC) in Iberian red deer during and out of the breeding season 
Eliana Pintus, José Luis Ros-Santaella and José Julián Garde

Few studies have dealt with spermatogenesis in red deer, which represent a good model in which to investigate testicular function owing to their reproductive seasonality. Herein, we found that both spermatogenesis and Sertoli cell number decreased significantly out of the breeding season in Iberian red deer. Our results provide new insights into the dynamics of spermatogenesis and Sertoli cell populations, and challenge current dogma that Sertoli cell number is fixed in adulthood.

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Published online 28 February 2014
Sperm survival kinetics in different types of bull semen: progressive motility, plasma membrane integrity, acrosomal status and reactive oxygen species generation 
Mushtaq Ahmad, Nasim Ahmad, Amjad Riaz and Muhammad Anzar

Sperm survival is important for the fertilisation of oocytes. The kinetics of progressive sperm motility, acrosomes, plasma membrane and generation of reactive oxygen species were determined in vitro over 24 h. Bull sperm longevity was poor in frozen–thawed semen. Different types of spermatozoa followed different death pathways. All types of spermatozoa generated superoxide anions during incubation, which adversely affected their plasma membranes.

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Published online 21 February 2014
Effect of different freezing velocities on the quality and fertilising ability of cryopreserved rabbit spermatozoa 
E. Mocé, E. Blanch, A. Talaván and M. P. Viudes de Castro

The freezing step of the protocol for rabbit sperm cryopreservation negatively influences the performance of the spermatozoa. In this study we show the important role that the freezing rate plays in the quality and fertilising ability of the spermatozoa, with medium freezing rates (–40°C min–1) more suitable for freezing rabbit spermatozoa. These results could help to develop reliable protocols for rabbit sperm cryopreservation.

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Published online 19 February 2014
Elective single versus double embryo transfer: live birth outcome and patient acceptance in a prospective randomised trial 
Nicolás Prados, Rocío Quiroga, Cinzia Caligara, Myriam Ruiz, Víctor Blasco, Antonio Pellicer and Manuel Fernández-Sánchez

The aim of the study was to determine which strategy (i.e. single (eSET) versus double (eDET) embryo transfer) has a better trade-off in live birth delivery considering patient acceptance. The live birth delivery rate was significantly higher after eDET versus eSET (65% vs 42%, respectively), but no multiple births were observed after eSET compared with 35% after eDET. Nearly half the patients refused eSET even after having been well informed of its benefits.

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Published online 19 February 2014
Oocyte expression, secretion and somatic cell interaction of mouse bone morphogenetic protein 15 during the peri-ovulatory period 
Brigitta Mester, Lesley J. Ritter, Janet L. Pitman, Adrian H. Bibby, Robert B. Gilchrist, Kenneth P. McNatty, Jennifer L. Juengel and C. Joy McIntosh

New work investigating how mammalian eggs achieve ovulation has found that the key fertility protein bone morphogenetic protein 15 (BMP15) is produced by the egg in more than one protein form just before ovulation. Whereas most previous studies focused on the mature region of BMP15, the present study shows that the proregion form is also increasingly produced just before ovulation and binds to surrounding cumulus and granulosa cells. This finding furthers our understanding of the complexity of BMP15 signalling and provides promising new routes of improving fertility treatments.

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blank image Reproduction, Fertility and Development
Volume 27 Number 4 2015
Oocyte and Embryo Metabolism

 
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A developmental tale – metabolism takes centre stage 
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Roger Sturmey and Rebecca Krisher
pp. iii-iv
 
 

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History of oocyte and embryo metabolism 
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Henry J. Leese
pp. 567-571

This review considers the forces that have driven research on egg and embryo metabolism. An early focus on basic research was followed by a period of neglect due to overemphasis on improving assisted conception techniques. Metabolism is now back centre stage and an integral part of exciting research to understand how embryos develop, with the goal of ensuring the ‘best start in life’.

 
  
 

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Nutrient pathways regulating the nuclear maturation of mammalian oocytes 
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Stephen M. Downs
pp. 572-582

Proper energy provision is the lifeblood of oocyte maturation that yields fertilisable ova with full developmental potential. This process involves complex interaction between carbohydrate, purine and fatty acid metabolic pathways fuelling nuclear maturation to MII and the cytoplasmic maturation necessary for developmental competence. An understanding of such metabolic relationships enables the optimisation of oocyte maturation media necessary to achieve these developmental endpoints.

 
  
 

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Hyperglycaemia and lipid differentially impair mouse oocyte developmental competence 
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Siew L. Wong, Linda L. Wu, Rebecca L. Robker, Jeremy G. Thompson and Melanie L. Sutton McDowall
pp. 583-592

This study investigated the influence of high glucose and lipid levels (as seen in obesity) on oocyte health. Both lipid and glucose treatments compromised embryo development and increased the activities of glucose fuel sensing (hexosamine biosynthetic pathway) and lipotoxicity responses (endoplasmic reticulum-stress, ER-stress). However, there were no additive effects when glucose and lipids were combined, and in the case of ER-stress markers, glucose partially recovered the effects of lipotoxicity.

 
  
 

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Distinct localisation of lipids in the ovarian follicular environment 
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Tawiwan Pantasri, Linda L. Wu, M. Louise Hull, Thomas R. Sullivan, Michael Barry, Robert J. Norman and Rebecca L. Robker
pp. 593-601

Obesity is associated with compromised oocyte quality. This study examined lipids within human follicular fluid and oocytes to determine whether they are altered by obesity or associated with oocyte quality. The results show that the ovarian follicular environment, which surrounds and nourishes the oocyte, exhibits a distinct metabolite profile compared to blood; but do not reveal associations between these parameters on and IVF outcomes

 
    | Supplementary Material (296 KB)
 

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Applying metabolomic analyses to the practice of embryology: physiology, development and assisted reproductive technology 
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Rebecca L. Krisher, Adam L. Heuberger, Melissa Paczkowski, John Stevens, Courtney Pospisil, Randall S. Prather, Roger G. Sturmey, Jason R. Herrick and William B. Schoolcraft
pp. 602-620

Metabolomics technology permits non-invasive monitoring of multiple metabolites from a single embryo during in vitro culture. Applying metabolomics to the analysis of individual embryos from several species, we demonstrate species- and stage-specific differences in metabolism and significant metabolic differences between in vivo- and in vitro-produced embryos. These results provide an insight into the metabolic pathways used by embryos, and their relationship with embryo quality.

 
  
 

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Ambient ionisation mass spectrometry for lipid profiling and structural analysis of mammalian oocytes, preimplantation embryos and stem cells 
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Christina R. Ferreira, Alan K. Jarmusch, Valentina Pirro, Clint M. Alfaro, Andres F. González-Serrano, Heiner Niemann, Matthew B. Wheeler, Rathnaweera A. C. Rabel, Judy E. Hallett, Rebecca Houser, Annemarie Kaufman and R. Graham Cooks
pp. 621-637

Lipid structural characterisation in single oocytes and preimplantation embryos is relevant for studies on cryopreservation, nutrition and the impact of culture on embryonic metabolism. In this paper, research on lipidomics of single oocytes and preimplantation embryos by ambient mass spectrometry is reviewed and discussed. This approach alone or in combination with other techniques can have a profound impact on the understanding of developmental biology and cell differentiation.

 
    | Supplementary Material (1.3 MB)
 

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Blastocyst metabolism 
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David K. Gardner and Alexandra J. Harvey
pp. 638-654

Perturbations in blastocyst metabolism compromise development of the subsequent fetus. Nutrient availability, oxygen concentration and the redox state of the blastocyst all affect the relative activities of specific metabolic pathways. Significantly, metabolism regulates more than ATP generation, with metabolites and cofactors representing important regulators of the epigenome, putting metabolism centre stage when considering interactions of the blastocyst with its environment.

 
  
 

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Arginine increases development of in vitro-produced porcine embryos and affects the protein arginine methyltransferase–dimethylarginine dimethylaminohydrolase–nitric oxide axis 
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Bethany K. Redel, Kimberly J. Tessanne, Lee D. Spate, Clifton N. Murphy and Randall S. Prather
pp. 655-666

In vitro culture results in porcine embryos that are less developmentally competent compared to in vivo-produced embryos. Supplementing culture medium with an essential amino acid, arginine, improves embryo development in vitro and by affecting the NO–DDAH–PRMT axis. A better understanding of the metabolic response to additional arginine will help to improve the production of pig embryos for agriculture and biomedicine.

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One-carbon metabolism and epigenetic regulation of embryo development 
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Juan Xu and Kevin D. Sinclair
pp. 667-676

Deficiencies in one-carbon related substrates and/or cofactors, such as folate and vitamin B12, during the periconceptional period can lead to epigenetic alterations in DNA and associated proteins in developmentally important genes in the embryo. Such epigenetic modifications can negatively impact on the subsequent health of offspring. Appropriate parental nutrition during this period can obviate these effects, particularly in subjects carrying risk alleles.

 
  
 

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Nutritional supplementation and artificial reproductive technique (ART) outcomes 
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A. J. Kermack and N. S. Macklon
pp. 677-683

Couples wishing to improve their chances when embarking on treatment with artificial reproductive technologies may turn to adjuvant therapies such as nutritional supplements. However, the practice of routine dietary supplementation prior to, and during in vitro fertilisation (IVF) treatment has not been subject to well-powered prospective randomised trials. Although the importance of nutrition in early pregnancy has been determined, further work examining specific nutrients is required before clinical advice can be given.

 
  
 

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Do little embryos make big decisions? How maternal dietary protein restriction can permanently change an embryo’s potential, affecting adult health 
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Tom P. Fleming, Adam J. Watkins, Congshan Sun, Miguel A. Velazquez, Neil R. Smyth and Judith J. Eckert
pp. 684-692

Poor maternal nutrition around conception permanently changes embryo development, affecting growth and metabolism into adulthood, increasing chronic disease risk. Evidence suggests preimplantation embryos ‘assess’ maternal nutrient quality and ‘decide’ the optimal ‘strategy’ for emerging extra-embryonic lineages to improve nutrient delivery and protect the competitive fitness of offspring. However, if postnatal nutrition is plentiful, such adaptations promote adult cardiometabolic disease, a legacy from our beginnings, with healthcare implications.

 
  
 

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Nutrition and maternal metabolic health in relation to oocyte and embryo quality: critical views on what we learned from the dairy cow model 
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Jo L. M. R. Leroy, Sara D. M. Valckx, Lies Jordaens, Jessie De Bie, Karolien L. J. Desmet, Veerle Van Hoeck, Jack H. Britt, Waleed F. Marei and Peter E. J. Bols
pp. 693-703

Nutrition and maternal metabolic health affect oocyte and embryo quality. This review summarises the value of in vivo and in vitro animal models to explain decreased oocyte and embryo quality in metabolically compromised individuals, with an emphasis on the bovine model. The challenges of current in vitro models are highlighted and the difficulties in extrapolating in vitro results to the in vivo situation are discussed.

 
  
 

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Dexamethasone treatment of pregnant F0 mice leads to parent of origin-specific changes in placental function of the F2 generation 
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O. R. Vaughan, H. M. Phillips, A. J. Everden, A. N. Sferruzzi-Perri and A. L. Fowden
pp. 704-711

Antenatal treatment of mothers with synthetic glucocorticoids like dexamethasone improves neonatal viability in preterm human infants but reduces birth weight and programs life-long, detrimental cardiometabolic traits in experimental animals. This study demonstrates that placental function is altered in the offspring of female mice that were overexposed to dexamethasone before birth by treatment of their mothers. The findings suggest that the placenta can contribute to the intergenerational inheritance of phenotypic traits induced by grandparental glucocorticoid exposure.

 
  
 

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Embryonic health: new insights, mHealth and personalised patient care 
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Régine P. M. Steegers-Theunissen and Eric A. P. Steegers
pp. 712-715

Embryonic health is a new theme in research and healthcare. We emphasise the importance of lifestyle of parents-to-be, which significantly contributes to embryonic health with short-term health consequences for reproduction and pregnancy, and long term for health during the life course. These individuals and healthcare professionals therefore should be adopting a life course approach using personalised lifestyle care by mHealth pregnancy platforms, such as www.slimmerzwanger.nl.

 
  
 

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Adverse effects of obesity and/or high-fat diet on oocyte quality and metabolism are not reversible with resumption of regular diet in mice 
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Kasey A. Reynolds, Anna L. Boudoures, Maggie M.-Y. Chi, Qiang Wang and Kelle H. Moley
pp. 716-724

Obesity is a global epidemic and has a significant adverse effect on human reproduction. The hypothesis of this study was that obesity impacts oocyte metabolism and its ability to undergo meiosis, but that this process may be reversible with dietary modification and weight reduction. Unfortunately in this model, these “life style” interventions did not reverse the adverse effect of diet and obesity on oocyte quality.

 
  
 

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These articles have been peer reviewed and accepted for publication. They are still in production and have not been edited, so may differ from the final published form.

    RD15131  Accepted 21 May 2015
    Effects of brain-derived neurotrophic factor on oocyte maturation and embryonic development in a rat model of polycystic ovary syndrome
    Qiaoli Zhang, Dong Liu, Meiling Zhang, Na Li, Shulan Lu, Yanzhi Du, Zijiang Chen
    Abstract


    RD14290  Accepted 22 May 2015
    Testicular injection of busulfan for recipient preparation in transplantation of spermatogonial stem cells in mice
    Qin Yusheng, Ling Liu, Yanan He, Wenzhi Ma, Huabin Zhu, Mingyuan Liang, Haisheng Hao, Tong Qin, Xueming Zhao, Dong Wang
    Abstract


    RD15101  Accepted 20 May 2015
    Extended ex-vivo culture of fresh and cryopreserved whole sheep ovaries
    Sara Maffei, Giovanna Galeati, Georgia Pennarossa, Tiziana Brevini, Fulvio Gandolfi
    Abstract


    RD14348  Accepted 14 May 2015
    Lactation-induced changes in metabolic status and follicular fluid metabolomic profile in post-partum dairy cows
    Niamh Forde, Aoife O'Gorman, Helena Whelan, Patrick Duffy, Lydia O'Hara, Alan Kelly, Vitezslav Havlicek, Urban Besenfelder, Lorraine Brennan, Patrick Lonergan
    Abstract


    RD14398  Accepted 06 May 2015
    Interaction of the TGF-β and Notch signaling pathways in the regulation of granulosa cell proliferation
    Xifeng Zhang, Xiaofeng Sun, Xinghong Sun, Shun-Feng Cheng, Junjie Wang, Yanni Feng, Yong Zhao, Shen Yin, Zhumei Hou, Wei Shen
    Abstract


    RD15092  Accepted 05 May 2015
    Androgen and Estrogen modulation by D-Aspartate in Rat Epididymis
    Sara Falvo, Maria Maddalena Di Fiore, Lavinia Burrone, Gabriella Chieffi Baccari, Salvatore Longobardi, Alessandra Santillo
    Abstract


    RD14408  Accepted 29 April 2015
    Activation of TRAIL receptor genes expression following DNA demethylation in placental choriocarcinoma and transformed cell lines
    Yubin Ding, Pan Wu, Mei Chen, Qing Liu, Lin He, Qian Feng, Xi Lan, Xue Zhang, Qing Geng, Xiong Wang
    Abstract


    RD14384  Accepted 27 April 2015
    Plasma prolactin concentrations during lactation, pouch young development and the return to behavioural oestrus in captive koalas (Phascolarctos cinereus)
    Katie Ballantyne, Stephen Anderson, Al Mucci, Vere Nicolson, Stephen Johnston
    Abstract


    RD15022  Accepted 26 April 2015
    Head morphology of ram spermatozoa is associated with their ability to migrate in-vitro and correlates with fertility
    Carmen Martínez-Rodríguez, Mercedes Álvarez, Elena López-Urueña, Susana Gomes-Alves, Luis Anel-López, Jorge Tizado, Luis Anel, Paulino Paz
    Abstract


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


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


    RD14211  Accepted 02 December 2014
    Breed specific factors influence embryonic lipid composition: comparison between Jersey and Holstein
    Luis Baldoceda Baldeon, Isabelle Gilbert, Dominic Gagne, Christian Vigneault, Patrick Blondin, Christina Ferreira, Claude Robert
    Abstract


    RD12339  Accepted 19 October 2012
    The critical importance of ovarian angiogenesis
    Bob Robinson
    Abstract


13


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1. Published 4 December 2014
IETS 2015 abstracts


2. Published 4 December 2014
Gamete derivation from embryonic stem cells, induced pluripotent stem cells or somatic cell nuclear transfer-derived embryonic stem cells: state of the art

Charles A. Easley, Calvin R. Simerly and Gerald Schatten

3. Published 4 December 2014
Exploring the function of long non-coding RNA in the development of bovine early embryos

Julieta Caballero, Isabelle Gilbert, Eric Fournier, Dominic Gagné, Sara Scantland, Angus Macaulay and Claude Robert

4. Published 4 December 2014
Epigenetics, embryo quality and developmental potential

Nathalie Beaujean

5. Published 4 December 2014
Genetic tools to improve reproduction traits in dairy cattle

A. Capitan, P. Michot, A. Baur, R. Saintilan, C. Hozé, D. Valour, F. Guillaume, D. Boichon, A. Barbat, D. Boichard, L. Schibler and S. Fritz

6. Published 4 December 2014
Genes and environmental factors that influence disease resistance to microbes in the female reproductive tract of dairy cattle

I. M. Sheldon

7. Published 19 June 2014
Replacing serum in culture medium with albumin and insulin, transferrin and selenium is the key to successful bovine embryo development in individual culture

E. Wydooghe, S. Heras, J. Dewulf, S. Piepers, E. Van den Abbeel, P. De Sutter, L. Vandaele and A. Van Soom

8. Published 4 December 2014
Genetic variation in resistance of the preimplantation bovine embryo to heat shock

Peter J. Hansen

9. Published 4 December 2014
Metabolic determinants of embryonic development and stem cell fate

Clifford D. L. Folmes and Andre Terzic

10. Published 17 July 2014
MicroRNA-regulated molecular mechanism underlying bovine subclinical endometritis

D. Hailemariam, S. Ibrahim, M. Hoelker, M. Drillich, W. Heuwieser, C. Looft, M. U. Cinar, E. Tholen, K. Schellander and D. Tesfaye

11. Published 22 August 2014
Pentose phosphate pathway activity: effect on in vitro maturation and oxidative status of bovine oocytes

Cynthia Gutnisky, Gabriel C. Dalvit, Jeremy G. Thompson and Pablo D. Cetica

12. Published 17 April 2015
Nutrition and maternal metabolic health in relation to oocyte and embryo quality: critical views on what we learned from the dairy cow model

Jo L. M. R. Leroy, Sara D. M. Valckx, Lies Jordaens, Jessie De Bie, Karolien L. J. Desmet, Veerle Van Hoeck, Jack H. Britt, Waleed F. Marei and Peter E. J. Bols

13. Published 17 April 2015
History of oocyte and embryo metabolism

Henry J. Leese

14. Published 17 April 2015
Hyperglycaemia and lipid differentially impair mouse oocyte developmental competence

Siew L. Wong, Linda L. Wu, Rebecca L. Robker, Jeremy G. Thompson and Melanie L. Sutton McDowall

15. Published 23 January 2015
Tolerance of spermatozoa to hypotonic stress: role of membrane fluidity and correlation with cryosurvival

Harriëtte Oldenhof, Anna Heutelbeck, Anne-Kathrin Blässe, Heinrich Bollwein, Gunilla Martinsson, Willem F. Wolkers and Harald Sieme

16. Published 22 August 2014
Three-dimensional systems for in vitro follicular culture: overview of alginate-based matrices

Ivina R. Brito, Isadora M. T. Lima, Min Xu, Lonnie D. Shea, Teresa K. Woodruff and José R. Figueiredo

17. Analysis and characterisation of bovine oocyte and embryo biomarkers by matrix-assisted desorption ionisation mass spectrometry imaging

Roseli F. Gonçalves, Mónica S. Ferreira, Diogo N. de Oliveira, Rafael Canevarolo, Marcos A. Achilles, Daniela L. D'Ercole, Peter E. Bols, Jose A. Visintin, Gary J. Killian and Rodrigo R. Catharino

18. Published 17 April 2015
Nutrient pathways regulating the nuclear maturation of mammalian oocytes

Stephen M. Downs

19. Published 11 March 2015
In vitro manipulation techniques of porcine embryos: a meta-analysis related to transfers, pregnancies and piglets

Ying Liu, Juan Li, Peter Løvendahl, Mette Schmidt, Knud Larsen and Henrik Callesen

20. Published 4 December 2014
Reproductive performance: at the cross-road of genetics, technologies and environment

Daniel Vaiman


      
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