title: Generation of parthenogenetic goat blastocysts: effects of different activation methods and culture media.
abstract: 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.
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Hruda Nanda
authors.initials: HN
authors.lastname: Malik
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Dinesh Kumar
authors.initials: DK
authors.lastname: Singhal
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Shrabani
authors.initials: S
authors.lastname: Saugandhika
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Amit
authors.initials: A
authors.lastname: Dubey
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Ayan
authors.initials: A
authors.lastname: Mukherjee
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Raxita
authors.initials: R
authors.lastname: Singhal
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Sudarshan
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Jai Kumar
authors.initials: JK
authors.lastname: Kaushik
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Ashok Kumar
authors.initials: AK
authors.lastname: Mohanty
authors.affiliation: Indian Veterinary Research Institute,Veterinary Physiology and Climatology,Izatnagar,Bareilly,India.
authors.forename: Bikash Chandra
authors.initials: BC
authors.lastname: Das
authors.affiliation: Indian Veterinary Research Institute,Veterinary Physiology and Climatology,Izatnagar,Bareilly,India.
authors.forename: Sadhan
authors.initials: S
authors.lastname: Bag
authors.affiliation: Central Avian Research Institute,Poultry Housing and Management,Izatnagar,Bareilly,India.
authors.forename: Subrata Kumar
authors.initials: SK
authors.lastname: Bhanja
authors.affiliation: Animal Biotechnology Center,National Dairy Research Institute,Karnal,India.
authors.forename: Dhruba
authors.initials: D
authors.lastname: Malakar
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000502:physiology; D061207:Calcium Ionophores / Q000494:pharmacology; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006041:Goats; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D010312:Parthenogenesis / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D061207:Calcium Ionophores; D003470:Culture Media; D000431:Ethanol
keywords: Parthenogenetic activation