2-Title: Generation of parthenogenetic embryos by ethanol activation in buffalo

Authors: Gopal Puri and Sadhan Bag

Source: Ruminant Science (2014)-3(1):5-7.

How to cite this manuscript: Puri Gopal and Bag Sadhan (2014). Generation of parthenogenetic embryos by ethanol activation in buffalo. Ruminant Science 3(1):5-7.


The present experiment was carried out to generate parthenogenetic embryo by ethanol activation of oocytes in buffalo. Oocytes were isolated from ovaries brought from local slaughter house. Isolated good quality of oocytes were matured in maturation media and kept in CO2 incubator for 24 hrs. The matured oocyte were activated by exposure to 7% ethanol for 5 minutes followed by incubation in 2mM 6-DMAP for 4 hr. After incubation in 6-DMAP, activated oocytes were finally shifted to mSOF media and kept in CO2 incubator for 24 hrs. The cleavage rate of oocytes was observed after 48-72 hr and 7 days. A total no. of 124 oocytes was used for ethanol activation and subsequent cleavage rate of activated oocytes were recorded. Result showed that overall cleaving percentage of oocytes was 49.80% and the development of embryos of 2 cell, 4 cell, 8 cell, morulae and blastocyst stages were 8.06%, 17.74%, 16.12%, 7.25% and 1.61%, respectively. It can be concluded that parthenogenetic embryos leads to haploid embryo production but fail to develop to term and exhibit phenotypic abnormalities might be due to inherently limited in their developmental capacity.


Alberio R, Zakhartchenko V, Motlik J and Wolf E (2001). Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. International Journal of Developmental Biology 45(7):797-809.

Bogliolo L, Calvia P, Leoni G, Loi P, Ledda S and Moor RM (1996).  Uncoupling of histone Hl activity from cell cycle progression in parthenogenetically activated sheep oocytes. In: Proceedings of the Society for the Study of Fertility Annual Meeting; Nottingham, UK. Abstract 39.

Gasparrini B, Boccia L, De Rosa A, Di Palo R, Campanile G and Zicarelli L (2004). Chemical activation of buffalo (Bubalus bubalis) oocytes by different methods. Theriogenology 62(9):1627-1637.

Kaufman MH (1984). Early mammalian development: Parthenogenetic studies. Cambridge University Press, Cambridge.

Kaufman MH, Huberman E and Sachs L (1975). Genetic control of haploid parthenogenetic development in mammalian embryos. Nature 254:694-705.

Kobayashi K, Yamashita S and Hoshi H (1994). Influence of epidermal growth factor and transforming growth factor-alpha on in vitro maturation of cumulus cell-enclosed bovine oocytes in a defined medium. Journal of Reproduction and Fertility 100(2):439-446,

Kono T (2006). Genomic imprinting is a barrier to parthenogenesis in mammals. Cytogenetic Genome Research 113:31-35.

Ledda S, Loi P, Fulka J and Jr Moor RM (1996).  The effect of 6-dimethylaminopurine on DNA synthesis in activated mammalian oocytes.  Zygote 4:7-9.

McGrath J and Solter D (1984). Completion of mouse embryogenesis requires both the maternal and paternal genomes. Cell 37:179-183.

Presicce GA and Yang X (1994). Nuclear dynamics of parthenogenesis of bovine oocytes matured in vitro for 20 and 40 h and activated with combined ethanol and cycloheximide treatment. Molecular Reproduction Development 37:61-68.

Ranjan R, Singh  RK Yasotha T, Manish Kumar,  Gopal Puri,  Kuldeep Kumar, Renu Singh,  Sanjeev Bhure,  Malakar D, Bhanja SK, Sarkar M, Das  BC and Sadhan Bag (2013).  Effect of actin polymerization inhibitor during oocyte maturation on parthenogenetic embryo development and ploidy in capra hircus. Biochemical Genetics 51(11-12):944-953

Szollosi MS, Kubiak JZ, Debey P, de Pennart H, Szollosi D and Maro B (1993). Inhibition of protein kinases by 6-dimethylaminopurine accelerates the transition to interphase in activated mouse oocytes. Journal of Cell Science 104:861-872.