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Generation of parthenogenetic goat blastocysts: effects of different activation methods and culture media

Published online by Cambridge University Press:  10 January 2014

Hruda Nanda Malik
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Dinesh Kumar Singhal
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Shrabani Saugandhika
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Amit Dubey
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Ayan Mukherjee
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Raxita Singhal
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Sudarshan Kumar
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Jai Kumar Kaushik
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Ashok Kumar Mohanty
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
Bikash Chandra Das
Affiliation:
Indian Veterinary Research Institute, Veterinary Physiology and Climatology, Izatnagar, Bareilly, India
Sadhan Bag
Affiliation:
Indian Veterinary Research Institute, Veterinary Physiology and Climatology, Izatnagar, Bareilly, India
Subrata Kumar Bhanja
Affiliation:
Central Avian Research Institute, Poultry Housing and Management, Izatnagar, Bareilly, India
Dhruba Malakar*
Affiliation:
Animal Biotechnology Center, National Dairy Research Institute, Karnal, India
*
All correspondence to: Dhruba Malakar. Animal Biotechnology Center, National Dairy Research Institute, Karnal, India. Tel: +91 9416741839. e-mail: [email protected]

Summary

The present study was carried out to investigate the effects of different activation methods and culture media on the in vitro development of parthenogenetic goat blastocysts. Calcium (Ca2+) ionophore, ethanol or a combination of the two, used as activating reagents, and embryo development medium (EDM), modified Charles Rosenkrans (mCR2a) medium and research vitro cleave (RVCL) medium were used to evaluate the developmental competence of goat blastocysts. Quantitative expression of apoptosis, stress and developmental competence-related genes were analysed in different stages of embryos. In RVCL medium, the cleavage rate of Ca2+ ionophore-treated oocytes (79.61 ± 0.86) was significantly (P < 0.05) higher than in ethanol (74.90 ± 1.51) or in the combination of both Ca2+ ionophore and ethanol. In mCR2a or EDM, hatched blastocyst production rate of Ca2+ ionophore-treated oocytes (8.33 ± 1.44) was significantly higher than in ethanol (6.46 ± 0.11) or in the combined treatment (6.70 ± 0.24). In ethanol, the cleavage, blastocyst and hatched blastocyst production rates in RVCL medium (74.90 ± 1.51, 18.30 ± 1.52 and 8.24 ± 0.15, respectively) were significantly higher than in EDM (67.81 ± 3.21, 14.59 ± 0.27 and 5.59 ± 0.42) or mCR2a medium (65.09 ± 1.57, 15.36 ± 0.52 and 6.46 ± 0.11). The expression of BAX, Oct-4 and GlUT1 transcripts increased gradually from 2-cell stage to blastocyst-stage embryos, whereas the transcript levels of Bcl-2 and MnSOD were significantly lower in blastocysts. In addition, different activation methods and culture media had little effect on the pattern of variation and relative abundance of the above genes in different stages of parthenogenetic activated goat embryos. In conclusion, Ca2+ ionophore as the activating agent, and RVCL as the culture medium are better than other tested options for development of parthenogenetic activated goat blastocysts.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2014 

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