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The present study aimed to investigate whether follicular fluid, gonadotropins (FSH, LH and estradiol-17β), and combination of both types of supplementation (follicular fluid + gonadotropins) in maturation medium influence the transcript abundance of germ cell marker genes ( MATER , ZAR1 , GDF9 and BMP15 ) in goat oocytes. Grade I and II oocytes were collected from the goat ovaries and matured further under in-vitro conditions using four different maturation regimens viz , group B containing basal media (TCM 199 + BSA + serum), group C (basal media + 10% follicular fluid), group D (basal media + FSH + LH + estradiol-17β) and group E (basal media + 10% follicular fluid + FSH + LH + estradiol-17β). Group A consisted of immature oocytes. Expression profile of MATER, ZAR1, GDF9, and BMP15 was analyzed in matured oocytes of different groups as well as immature oocytes using real-time PCR. The relative abundance of MATER gene in immature oocytes (group A) was found to be significantly higher (p ZAR1 was found to be significant different (P GDF9 and BMP15 was significantly higher (P MATER and ZAR1 transcript were down regulated after maturation; however BMP15 and GDF9 transcripts were upregulated after maturation. For better in vitro embryo production (IVEP) out comes goat oocytes may be matured in vitro in TCM-199 supplemented with follicular fluid and gonadotropins.

The objective of this study was to compare the effectiveness of different activation treatments of in vitro matured oocytes and their developmental potency in potassium simplex optimization medium. Ovaries were collected from the local abattoir and transported within 4 h to the laboratory in warm saline (37 °C) containing 100 IU penicillin-G and 100 μg streptomycin sulphate per ml. A total of 1004 cumulus oocyte complexes (COC's) were collected from 454 ovaries. Oocytes were matured in TCM-199 medium containing FSH (5 μg/ml), LH (10 μg/ml), oestradiol-17β (1 μg/ml) supplemented with 10% fetal bovine serum and 3 mg/ml BSA at 38.5 °C and 5% CO2 in an incubator under humidified air for 27 h. After 27 h of IVM, oocytes were denuded, washed and selected 933 in vitro matured oocytes were randomly divided into four groups. Group 1 in vitro matured oocytes (n = 579), were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP for 4 h in KSOM. Group 2 in vitro matured oocytes (n = 145) were exposed to 7% ethanol for 5 min followed by treatment with 10 μg/ml CHX for 4 h in KSOM. Group 3 in vitro matured oocytes (n = 100) were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP and 10 μg/ml CHX for 4 h in KSOM. Group 4 in vitro matured oocytes (n = 109) were cultured for 4 h without any chemical activation treatment in KSOM medium (control). After 4 h of culture in different chemicals, the oocytes were washed five to ten times in the culture medium (KSOM) and cultured in 50 μl drops of KSOM. Development of activated oocytes was observed at every 48 h till day 10 post activation under an inverted phase contrast microscope (200x, Nikon, Japan). The cleavage rate in groups 1, 2, 3 and 4 were 42.83%, 58.62%, 74.0% and 0.00%, respectively and morula production in groups 1, 2 and 3 were 24.59%, 30.58% and 31.08%, respectively. These results indicated that the activation of in vitro matured oocytes by 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP and 10 μg/ml CHX for 4 h in KSOM is most favorable for parthenogenetic caprine embryos production.