title: Effect of Ca Ionophore On Blastocyst Production Following Intracytoplasmic Sperm Injection in Caprine Oocytes.
abstract: The aim of the present investigation was to study the effect of calcium ionophore activation on blastocyst production following intracytoplasmic sperm injection (ICSI) in in vitro-matured Caprine oocytes. A total of 470 in vitro-matured oocytes were selected and randomly divided in to three groups. Cumulus oocyte complexes (COCs) recovered by slicing the Caprine ovaries were matured in TCM199 supplemented with 10% foetal bovine serum (FBS) + 10% follicular fluid + FSH (5 μg/ml) + LH (10 μg/ml) + estradiol (1 μg/ml) + EGF (10 ng/ml) + BSA (3 mg/ml) for 27 h in humidified atmosphere at 38.5°C with 5% CO2 in CO2 incubator. After 27 h of culture, selected COCs (n = 470) were separated from cumulus cells by treating with 0.1% hyaluronidase enzyme and passing repeatedly through a fine pipette and randomly divided into three groups. In group 1, (n = 168) matured oocytes were injected with injection micropipette without sperm as control. In group 2, (n = 152) capacitated spermatozoa were injected into cytoplasm of in vitro-matured oocytes through injection micropipette. In group 3, (n = 150) capacitated spermatozoa were injected into cytoplasm of in vitro-matured oocytes through injection micropipette and then activated with 5 μm Ca ionophore for 5 min. The oocytes of all groups were then culture in RVCL media for embryo development. The cleavage rate was observed after 48-72 h of injection. The cleavage rate and blastocyst production in group 1, 2 and 3 were 0.00 and 0.00, 18.42 and 3.57 and 61.33% and 16.30%, respectively. The result indicated that mechanical activation failed to induce cleavage in in vitro-matured Caprine oocytes, whereas chemical activation of intracytoplasmic sperm-injected in vitro-matured Caprine oocytes showed significantly higher cleavage rate and blastocyst production as compare to non-activated oocytes.
authors.affiliation: ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
authors.forename: S D
authors.initials: SD
authors.lastname: Kharche
authors.affiliation: ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
authors.forename: J
authors.initials: J
authors.lastname: Pathak
authors.affiliation: ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
authors.forename: S
authors.initials: S
authors.lastname: Agarwal
authors.affiliation: ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
authors.forename: B
authors.initials: B
authors.lastname: Kushwah
authors.affiliation: ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
authors.forename: Aks
authors.initials: A
authors.lastname: Sikarwar
mesh_terms: D000818:Animals; D001755:Blastocyst; D061207:Calcium Ionophores / Q000494:pharmacology*; D005260:Female; D006041:Goats; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D008297:Male; D009865:Oocytes / Q000502:physiology; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*
chemical_list: D061207:Calcium Ionophores