Effect of cytochalasin B during oocyte maturation for parthenogenetic embryos generation in goat
250 / 50
Keywords:
Caprine, Cytochalasin B, Embryos, Ethanol, Maturation media, ParthenogenesisAbstract
The present experiment was conducted to study the effect of cytochalasin B (CCB) in maturation medium for parthenogenetic embryos generation in caprine (Capra hircus). The cumulus oocyte complexes (COC) were isolated, evaluated, graded and matured in maturation medium having different concentration of CCB (0 μg/ml, 5 μg/ml, 10 μg/ml, 15 μg/ml and 20 μg/ml). Matured oocytes were activated by 7% ethanol for 5 min followed by incubation with 2mM 6-dimethyl amino purine (DMAP) for 4 h and cultured in modified synthetic oviductal fluid (mSOF) in CO2 incubator at 37°C, 5% CO2, and 95% relative humidity. The cleavage rate and further embryo development was recorded after 48 h and then 96 h of in vitro culture. The overall oocyte recovery rate was 1.13±0.02/ovary. The overall cleavage rate was 77.22±1.04 and percentage of 2–4 cell, 8–16 and morula stage embryos production were 22.90±1.38, 34.46±0.89 and 42.64±1.04, respectively irrespective of different concentration of CCB used in maturation medium. There was nonsignificant difference of cleavage rate among different concentration of CCB used. The embryos development beyond 16 cell stage was higher in 15 μg/ml CCB compared to other concentration of CCB but nonsignificant compared to control (0 μg/ml CCB). The presence of CCB all throughout maturation (27 h) did not inhibit cleavage rate as well as embryo development, rather increased the embryo development beyond 16 cell stage.
Downloads
References
Deng M Q and Fan B Q. 1994. Intracellular free calcium changes of mouse oocytes during activation induced by ethanol or electrical stimulations and parthenogenetic development. Journal of Experimental Biology 27: 289-97.
Duarte G, Flores J A, Malpaux B and Delgadillo J A. 2008. Reproductive seasonality in female goats adapted to a subtropical environment persists independently of food availability. Domestic Animal Endocrinology 35: 362–70.
Fukui Y, Sawai K, Furudate M, Sato N, Iwasumi Y and Ohzaki K. 1992. Parthenogenetic development of bovine oocytes treated with ethanol and cytochalasin B after in vitro maturation. Molecular Reproduction and Development 33: 357–62.
Grupen C G, Nottle M B and Nagashima H. 2002. Calcium release at fertlization: artificial mimicking the oocyte response to sperm. Journal of Reproduction and Development 48: 313– 33.
Hanna B and Tarkowski A K. 1976. Diploid parthenogenetic mouse embryos produced by heat-shock and Cytochalasin B. Journal of Embryology and Experimental Morphology 35(l): 25–39.
Jacek K, Andras P, Michele W and Bernard M. 1991. Genetically identical parthenogenetic mouse embryos produced by inhibition of the first meiotic cleavage with cytochalasin D. Development 111: 763–69.
Jones K T, Carroll J and Whittingham D G. 1995. Ionomycin, thapsigargin, ryanodine, and sperm induced Ca2+ release increase during meiotic maturation of mouse oocytes. Journal of Biological Chemistry 270: 6671–77.
Kim N H, Moon S J, Prather R S and Day B N. 1996. Cytosketelal alteration in aged porcine oocytes and parthenogenesis. Molecular Reproduction and Development 43: 513–18.
Kobayashi K, Yamashitu S and Hushi H. 1994. Influence of epidermal growth factors and transforming growth factors on in vitro maturation of cumulus enclosed bovine oocytes in defined medium. Journal of Reproduction and Infertility 12: 439–46.
Lan G, Dong H, Yan-Guanng W, Zheng-Bin H, Suo-Feng M, Xin- Yong L, Chong-Le C and Jing H T. 2005. Effects of duration, concentration, and timing of ionomycin and 6- dimethylaminopurine (6-DMAP) treatment on activation of goat oocytes. Molecular Reproduction and Development 71: 380–88.
Liu L, Ju J C and Yang X. 1998. Parthenogenetic development and protein patterns of newly matured bovine oocytes after chemical activation. Molecular Reproduction and Development 49: 298–307.
Loi P, Ledda S, Fulka J Jr, Cappai P and Moor R M. 1998. Development of parthenogenetic and cloned ovine embryos: effect of activation protocols. Biology of Reproduction 58: 1177–87.
Mishra V, Misra A K and Sharma R A. 2008. Comparative study of parthenogenic activation and in vitro fertilization of bubaline oocytes. Animal Reproduction Science 103(3–4): 249–59.
Ongeri E M, Bormann C L, Butler R E, Melican D, Gavin W G, Echelard Y, Krisher R L and Behboodi E. 2001. Development of goat embryos after in vitro fertilization and parthenogenetic activation by different methods. Theriogenology 55: 1933– 45.
Pankaj R S, Satisha K B, Venkatesh V, Gupta R, Kumar K, Das B C, Majumdar A C, Bag S and Ranjan R. 2012. Effect of different activation protocol on generation of parthenogenetic embryos in caprine. Indian Journal of Animal Sciences 82 (11): 1323–26.
Ranjan R, Singh R K, Yasotha T, Kumar Manish, Puri Gopal, Kumar Kuldeep, Singh Renu, Bhure Sanjeev, Malakar D, Bhanja S. K, Sarkar M, Das B C and Bag Sadhan. 2013. Effect of actin polymerization inhibitor during oocyte maturation on parthenogenetic embryo development and their ploidy in Capra hircus. Biochemical genetics DOI 10.1007/S10528- 013–9619–4
Sathisha K B. 2009. ‘Studies on development of parthenogenetic embryos of different ploidy in buffalo.’ M V Sc Thesis. Indian Veterinary Research Institute, Deemed University, Izatnagar, India.
Venkatesh V. 2009. ‘Development of parthenogenetic stem cells of buffalo and their characterization with pluripotent markers.’ M V Sc Thesis. Indian Veterinary Research Institute, Deemed University, Izatnagar, India.
Yang X, Jiang S and Shi Z. 1992. Improved activation by combined cycloheximide and electrical pulse treatment of bovine follicular oocytes matured in vitro for 23–24 h. Biology of Reproduction 46: 117.
Yi Y J and Park C S. 2004. Parthenogenetic development of porcine oocytes treated by ethanol, cycloheximide, cytochalasin B and 6-dimethylaminopurine. Animal Reproduction Science 86: 297–304.
Young L E, Fernandes K, McEvoy T G, Butterwith S C, Gutierrez C G, Carolan C, Broadbent P J, Robinson J J, Wilmut I and Sinclair K D. 2003. Epigenetic change in IGF2R is associated with fetal overgrowth after sheep embryo culture. Nature Genetics 27: 153–54.
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2014 The Indian Journal of Animal Sciences

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.