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

Reproduction, Fertility and Development

Volume 25 Number 4 2013

RD11262Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro

Toshikiyo Takahashi, Yasushi Inaba, Tamas Somfai, Masahiro Kaneda, Masaya Geshi, Takashi Nagai and Noboru Manabe
pp. 589-599

In animal husbandry, freezing is widely used for the storage and transportation of embryos destined for transfer; however, high lipid content in embryos makes them sensitive to low temperatures. In this study, l-carnitine, an enhancer of lipid metabolism, has been successfully applied during embryo culture to improve development and freezing tolerance of bovine embryos. The use of l-carnitine offers a safe and non-invasive way to improve the efficacy of embryo-freezing technology in cattle.

RD11282Morphology, sex ratio and gene expression of Day 14 in vivo and in vitro bovine embryos

Grazieli M. Machado, Allice R. Ferreira, Monique M. Guardieiro, Michele R. Bastos, José O. Carvalho, Carolina M. Lucci, Tiago O. Diesel, Roberto Sartori, Rodolfo Rumpf, Maurício M. Franco and Margot A. N. Dode
pp. 600-608

The post hatching development (PHD) systems is a potential tool to verify the quality of in vitro produced embryos without transfer then for the recipient uterus. The study aimed to evaluate the efficiency of the system by comparing embryos produced in PHD system with the in vivo produced ones. The results showed that the system is not efficient enough to be use as a routine method for embryo evaluation.


As a valuable technology in assisted reproduction, egg freezing needs to be further evaluated, such as effects on cytoplasm quality and the nucleus. Through studying eggs frozen at different times after intracytoplasmic sperm injection or artificial activation, we found that differences in ploidy formation existed, whereas cytoskeleton assembly was not altered and there was no difference in blastocyst development compared with normal zygotes. We believe that proper freezing not only maintains the egg cytoskeleton but also contributes to eggs being able to maintain their reprogramming ability for spermatozoa during freezing, which could become a useful tool to monitor and optimize egg freezing.

RD12011Actin nucleator Arp2/3 complex is essential for mouse preimplantation embryo development

Shao-Chen Sun, Qing-Ling Wang, Wei-Wei Gao, Yong-Nan Xu, Hong-Lin Liu, Xiang-Shun Cui and Nam-Hyung Kim
pp. 617-623

Although it is well known that actin is involved in multiple processes during cell division, which also includes early embryo development, the molecular mechanism is still unclear. In the present study we found that inhibition of the actin nucleator Arp2/3 complex caused a decrease in actin within cells and the failure of embryo development. Our results indicate that the Arp2/3 complex is involved in mouse embryo development through the regulation of actin assembly.

RD12075Sperm capacitation combined with removal of the sperm acrosome and plasma membrane enhances paternal nucleus remodelling and early development of bovine androgenetic embryos

Yao Xiao, Hualin Zhang, Sibtain Ahmad, Liya Bai, Xiaomin Wang, Lijun Huo, Xin Zhang, Wengong Li, Xiang Li and Liguo Yang
pp. 624-638

The production of embryos using genetic material only from the father is difficult and the efficiency of this technique is extremely low in mammalian species. Mimicking several physiological processes during normal fertilisation, we treated bovine spermatozoa prior to the production of embryos and successfully improved their developmental capacity. Our results contribute to the understanding of the paternal contribution to embryonic development and to the production of transgenic animals.

RD12020Steroidogenesis in sheep pregnancy with intrauterine growth retardation by high-altitude hypoxia: effects of maternal altitudinal status and antioxidant treatment

Víctor H. Parraguez, Bessie Urquieta, Mónica De los Reyes, Antonio González-Bulnes, Susana Astiz and Andrés Muñoz
pp. 639-645

Fetal growth is adversely affected by environmental hypoxia and oxidative stress in sheep flocks kept at high altitudes. The administration of antioxidant vitamins may prevent these effects. Because placental steroidogenesis is a key factor for fetal growth, we evaluated the effects of supplementation with vitamins C and E on plasma concentrations of progesterone and 17β-oestradiol in high-altitude pregnancies in sheep. The concentrations of both hormones were higher in the supplemented groups, favouring fetal development and, thus, also improving sheep production at high altitudes.

RD11240Solea senegalensis vasa transcripts: molecular characterisation, tissue distribution and developmental expression profiles

Tiziana Pacchiarini, Ismael Cross, Ricardo B. Leite, Paulo Gavaia, Juan B. Ortiz-Delgado, Pedro Pousão-Ferreira, Laureana Rebordinos, Carmen Sarasquete and Elsa Cabrita
pp. 646-660

This research was focused on the Senegalese sole, a flatfish with high commercial value in Mediterranean aquaculture. The aim of this work was to characterise vasa gene to determine its usefulness as a molecular marker for germ cells during embryonic, larval development, juvenile and adulthood. We demonstrated that vasa products (mRNAs and proteins) allow germ cells identification establishing a suitable molecular tool for germinal cells biotechnology for Senegalese sole reproduction.

RD11271Exogenous long-term treatment with 17?-oestradiol alters the innervation pattern in pig ovary

Marlena Koszykowska, Jarosław Całka, Aleksandra Nidzgorska and Barbara Jana
pp. 661-673

The aim of this study was to determine the effect of long-term estradiol-17β (E2) exposure (simulation of pathological states occurring with estrogen overproduction) on innervation pattern of ovaries in adult gilts. In the ovaries of E2-injected gilts, the disturbances in the development of follicles, an increase in the total population of nerve fibers, including fibers containing DβH, NPY, and GAL were observed. These results suggest that hyperestrogenism caused by endogenous or exogenous estrogens may affect the innervation pattern of ovaries and consequently their function(s) and further support the importance of estrogens as modulators of neuronal plasticity.

RD12091The rat oocyte synthesises melatonin

Kenichiro Sakaguchi, Masanori T. Itoh, Noriyuki Takahashi, Wataru Tarumi and Bunpei Ishizuka
pp. 674-682

Melatonin, an indoleamine originally identified in the pineal gland, affects ovarian functions, and previous studies have suggested that melatonin is synthesised in the ovary. This study demonstrates that in rat oocytes, melatonin is synthesised from serotonin through the same pathway as in the pineal gland. Melatonin synthesised in the oocyte may be implicated in its own growth and/or maturation, for example, by acting as a calmodulin antagonist and/or an antioxidant.

RD12050Alpaca semen quality in relation to different diets

N. S. Juyena, J. Vencato, G. Pasini, I. Vazzana and C. Stelletta
pp. 683-690

Camelids are often wrongly considered as small bovine or big ovine in terms of nutritional requirements. Specific nutritional planes are essential for the reproductive management optimisation. Studies investigating the link between nutrition and reproductive performance are particularly needed in the male. The present study investigated the effect of different nutritional planes and found detrimental effects of energy excess on the biochemical characteristics of alpaca semen.

RD12138Involvement of microRNA-335-5p in cytoskeleton dynamics in mouse oocytes

Xiang-Shun Cui, Shao-Chen Sun, Yong-Kook Kang and Nam-Hyung Kim
pp. 691-699

The specific roles of microRNAs in terms of germ cell development are largely unknown. The present study found that spindle formation and the actin cap were affected after microinjection of miR-335-5p or its inhibitor during mouse oocyte maturation. On the basis of the results of the present study, it appears that microRNA mir-335-5p may be involved in cytoskeleton dynamics in mouse oocyte meiosis.

RD11221Employing mated females as recipients for transfer of cloned dog embryos

Geon A Kim, Hyun Ju Oh, Jung Eun Park, Min Jung Kim, Eun Jung Park, Sang Hyun Lim, Sung Keun Kang, Goo Jang and Byeong Chun Lee
pp. 700-706

Little is known about the use of mated bitches with in vivo-fertilised embryos as recipients for cloned embryos in dog cloning. Our study demonstrated successful production of cloned dogs with wild-type pups by using mated recipients as surrogate mothers. Thus, employing mated recipients could be a new method that may improve the efficiency of cloning in dogs and help with understanding the underlying mechanism of implantation.

RD12018Cryopreservation and long-term maintenance of bovine embryo-derived cell lines

Maryam Pashaiasl, Khodadad Khodadadi, Nadine M. Richings, Michael K. Holland and Paul J. Verma
pp. 707-718

The derivation of robust livestock embryonic stem cell lines, which promise a wide range of practical applications in agricultural biotechnology, including dissemination of elite genetics and sophisticated genome editing, has proven difficult to date. We examined the effect of small molecules that prevent differentiation of stem cells and methods of cryopreservation on long-term survival and retention of stem-cell characteristics of embryo-derived stem cells in the cow. We show that vitrification, which minimises cell damage, was effective for freezing cells, which could be maintained as stem cells for more than 240 days in culture.

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