2015The Indian Journal of Animal SciencesOpen access

Effect of protein phosphorylation inhibitor on production of parthenogenetic caprine embryos

S.D. Kharche, A. K. Goel, Swati Jindal, Puja Goel, Justin Kouamo, Sonia Sarswat

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Abstract

The objective of the present study was to compare the effect of different concentrations of 6-Dimethylaminopurine (6-DMAP) treatment in chemical activated oocytes to compare the developmental potency of parthenogenetic caprine embryos. Morphologicaly matured oocytes were denuded and activated with 5 μm calcium ionophore (ionomycin) for 5 min. Activated oocytes were randomly divided into 5 groups: Group 1 oocytes were cultured in modified Charles Rosenkrans medium (mCR2aa), without DMAP treatment, group 2 cultured in mCR2aa with 2.5 mM DMAP, group 3 cultured in mCR2aa with 5 mM DMAP, group 4 cultured in mCR2aa with 10 mM DMAP, group 5 cultured in mCR2aa with 20 mM DMAP for 4 h. The presumptive zygotes of 5 groups were washed and cultured in the embryo development medium. Development of parthenotes was observed at every 48 h till day 12 post insemination. The percentage of cleavage, morula, blastocyst and hatched blastocyst production in groups 1, 2, 3, 4 and 5 were 61.76, 6.67, 0.00 and 0.00; 59.79, 27.43, 10.62 and 0.00; 72.43, 41.62, 10.66 and 1.52; 64.61, 50.44, 7.83 and 0.00; 63.19, 19.42, 3.88 and 0.00% respectively. Results indicated that 5 mM DMAP treatment for chemically activated oocyte is the best treatment for development of parthenogenetic caprine embryos. In conclusion, it can be stated that for the production of caprine parthenogenetic embryos in vitro, concentration of protein phosphorylation inhibitor plays an important role in the activation treatment. Optimized activation protocols could enhance the production of viable parthenogenetic embryos.

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The objective of the present study was to compare the effect of different concentrations of 6-Dimethylaminopurine (6-DMAP) treatment in chemical activated oocytes to compare the developmental potency of parthenogenetic caprine embryos. Morphologicaly matured oocytes were denuded and activated with 5 μm calcium ionophore (ionomycin) for 5 min. Activated oocytes were randomly divided into 5 groups: Group 1 oocytes were cultured in modified Charles Rosenkrans medium (mCR2aa), without DMAP treatment, group 2 cultured in mCR2aa with 2.5 mM DMAP, group 3 cultured in mCR2aa with 5 mM DMAP, group 4 cultured in mCR2aa with 10 mM DMAP, group 5 cultured in mCR2aa with 20 mM DMAP for 4 h. The presumptive zygotes of 5 groups were washed and cultured in the embryo development medium. Development of parthenotes was observed at every 48 h till day 12 post insemination. The percentage of cleavage, morula, blastocyst and hatched blastocyst production in groups 1, 2, 3, 4 and 5 were 61.76, 6.67, 0.00 and 0.00; 59.79, 27.43, 10.62 and 0.00; 72.43, 41.62, 10.66 and 1.52; 64.61, 50.44, 7.83 and 0.00; 63.19, 19.42, 3.88 and 0.00% respectively. Results indicated that 5 mM DMAP treatment for chemically activated oocyte is the best treatment for development of parthenogenetic caprine embryos. In conclusion, it can be stated that for the production of caprine parthenogenetic embryos in vitro, concentration of protein phosphorylation inhibitor plays an important role in the activation treatment. Optimized activation protocols could enhance the production of viable parthenogenetic embryos.

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Available abstract

The objective of the present study was to compare the effect of different concentrations of 6-Dimethylaminopurine (6-DMAP) treatment in chemical activated oocytes to compare the developmental potency of parthenogenetic caprine embryos. Morphologicaly matured oocytes were denuded and activated with 5 μm calcium ionophore (ionomycin) for 5 min. Activated oocytes were randomly divided into 5 groups: Group 1 oocytes were cultured in modified Charles Rosenkrans medium (mCR2aa), without DMAP treatment, group 2 cultured in mCR2aa with 2.5 mM DMAP, group 3 cultured in mCR2aa with 5 mM DMAP, group 4 cultured in mCR2aa with 10 mM DMAP, group 5 cultured in mCR2aa with 20 mM DMAP for 4 h. The presumptive zygotes of 5 groups were washed and cultured in the embryo development medium. Development of parthenotes was observed at every 48 h till day 12 post insemination. The percentage of cleavage, morula, blastocyst and hatched blastocyst production in groups 1, 2, 3, 4 and 5 were 61.76, 6.67, 0.00 and 0.00; 59.79, 27.43, 10.62 and 0.00; 72.43, 41.62, 10.66 and 1.52; 64.61, 50.44, 7.83 and 0.00; 63.19, 19.42, 3.88 and 0.00% respectively. Results indicated that 5 mM DMAP treatment for chemically activated oocyte is the best treatment for development of parthenogenetic caprine embryos. In conclusion, it can be stated that for the production of caprine parthenogenetic embryos in vitro, concentration of protein phosphorylation inhibitor plays an important role in the activation treatment. Optimized activation protocols could enhance the production of viable parthenogenetic embryos.

Key concepts: Blastocyst, Embryo, Andrology, Ionomycin, Zygote, Parthenogenesis, Embryogenesis, Oocyte

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