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Vol. 9, Issue 9 (2020)

Efficacy of Ca ionophore and 6-dimethylaminopurine on activation of prepubertal ovine oocytes

Author(s):
Ramsingh L, KRC Reddy and Gopala Reddy A
Abstract:
The present study was carried out to compare the different activation protocols for production of parthenogenetic embryos from prepubertal sheep ovarian oocytes and to compare the in vitro fertilization derived embryos. In vitro fertilization (IVF) embryos were produced by adopting IVFMC of oocytes collected from prepubertal sheep ovaries from local slaughter house, Ziaguda (Hyderabad). The overall cleavage rate was 57.5 ± 0.38, 53 ± 0.87 and 8 ± 0.24%, respectively in treatments I (10% ESS + 10 µg/ml FSH + 10 µg/ml LH + µg/ml estradiol + 50 µg/ml gentamicin + TCM 199 B), II (10 µg/ml FSH + 10 µg/ml LH + µg/ml estradiol + 50 µg/ml gentamicin + TCM 199 B) and III (50 µg/ml gentamicin + TCM 199 B), respectively. The cleaved cells attained 2-cell, 4-cell, 8-cell and Morula stage were 38.5 ± 0.28, 27.25 ± 0.28, 24.5 ± 0.23 and 14 ± 0.16%, 31 ± 0.71, 20 ± 0.60, 11 ± 0.51 and 2 ± 0.21%,, 3 ± 0.24, 1 ± 0.20, 0 and 0%, respectively in treatments I, II and III. Among the three treatments, Treatment I achieved significantly (P<0.05) higher percentage of cleaved cells.
Parthenogenetic embryos were produced by using different concentrations of calcium ionophore + 6-dimethyl aminopurine (6-DMAP) in the concentrations of 1 µg/ml + 1 mM, 5 µg/ml + 2 mM and 10 µg/ml + 5 mM in protocol I, II and III. respectively and also a control was maintained using TCM 199 B + synthetic oviduct fluid (SOF). The results of embryonic development showed that there was significant (P<0.05) difference in cleavage rate between the activation treatments. Treatment II resulted in significantly (P<0.05) higher cleavage rate (66 ± 0.37%) followed by in vitro fertilization (IVF) (57.5 ± 0.38%), Treatment III (46.75 ± 0.62%) and Treatment I (38.5 ± 0.41%). No significant difference was observed between Treatment II and IVF pertaining to attainment of 2-cell stage, but differed significantly (P<0.05) when compared to Treatments I (21.75 ± 0.24%) and III (27.5 ± 0.32%). Pertaining to attainment of 4-cell stage, Treatment II resulted in significantly higher rate (30.25 ± 0.32%) followed by in-vitro fertilization (27.25 ±0.28%), Treatment III (23 ± 0.36%) and Treatment I (16.5 ± 0.18%). Pertaining to attainment of 8-cell stage, Treatment II resulted in significantly higher rate (26.75 ± 0.33%) followed by in-vitro fertilization (24.5± 0.23%), Treatment III (13.25± 3.20%) and Treatment I (15 ± 0.19%). Pertaining to attainment of Morula stage, Treatment II resulted in significantly higher rate (16.5 ± 0.23%) followed by in-vitro fertilization (14 ± 0.16%), Treatments III (12 ± 0.20%) and I (9.5± 0.16%).
Based on the results of the present study, it can be concluded that the exposure of in vitro mature oocytes to calcium ionophore at a concentration of 5 µg/ml for 5 min followed by 3 hrs incubation with 2 mM 6-DMAP (Protocol-II) was found to be the best for parthenogenetic embryo production in prepubertal ovine ovaries. Comparison of 2-cell stage between treatment II and IVF was similar. Other stages (4-cell, 8-cell and Morula stage) were significantly (p<0.05) higher in parthenogenetic activation of oocytes.
Pages: 216-219  |  479 Views  56 Downloads


The Pharma Innovation Journal
How to cite this article:
Ramsingh L, KRC Reddy, Gopala Reddy A. Efficacy of Ca ionophore and 6-dimethylaminopurine on activation of prepubertal ovine oocytes. Pharma Innovation 2020;9(9):216-219.

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