Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
Table of Contents
Reproduction, Fertility and Development

Reproduction, Fertility and Development

Volume 27 Number 8 2015

RD14049Impacts of and interactions between environmental stress and epigenetic programming during early embryo development

Michael J. Bertoldo, Yann Locatelli, Christopher O'Neill and Pascal Mermillod
pp. 1125-1136

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.

RD13265Effect 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
pp. 1137-1146

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.


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.


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.

RD14070Validation 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
pp. 1168-1174

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.

RD14024Open 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
pp. 1175-1180

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.


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.

RD14043Free-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
pp. 1187-1196

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.

RD14096Insulin 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
pp. 1197-1204

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.


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.

RD14113Transgenerational 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
pp. 1213-1221

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.

RD13436Low-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
pp. 1222-1232

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.


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.

RD14048Oocytes 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
pp. 1242-1248

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.

RD14091Dietary 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
pp. 1249-1261

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.

RD14381Activation 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
pp. 1262-1271

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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