Estrogen receptor gene 1 expression in caprine and its effect on fertility

The present study was undertaken to analyze the expression pattern of estrogen receptor 1 gene [ESR1] in Barbari bucks [fertile and non-fertile] identified on the basis of seminal quality traits and fertility trials. RNA was extracted from the spleen by Trizol method. The expression pattern of ESR1 gene was analyzed using real time polymerase chain reaction [RT-PCR]. The expression pattern of ESR1 gene was analyzed by RT-PCR [Roche LC-480]. Relative quantification by RT-PCR indicated that the ESR1 gene expression showed more fold in fertile bucks as compared to non-fertile

comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured caprine oocytes

The aim of the study was to compare the parthenogenetic activation and in vitro fertilization [IVF] of in vitro matured caprine oocytes. A total of 881 cumulus-oocyte complexes [COC's] were collected from 243 ovaries. Oocytes were matured in TCM-199 medium containing eCG [20 IU/ml], hCG [20 IUmicro g/ml], oestradiol-17beta [1 micro g/ml], BSA embryo tested [3 mg/ml] supplemented with 10% fetal bovine serum at 38.5[degree]C and 5% CO[2] in an incubator under humidified air for 27 h. Based on cumulus expansion, the maturation rate was 86.86%. Morphological matured oocytes [n=749] were selected, denuded and randomly divided into two groups. Group 1 [n=223] in vitro matured oocytes activated with 5 micro m calcium ionophore for 5 min and cultured in mCR[2]aa medium containing 5 mM DMAP for 4 h. After 4 h of DMAP treatment, the presumptive zygotes were washed and cultured in the embryo culture medium. Group 2 [n=526] in vitro matured oocytes processed for IVF in mTALP using fresh semen of a fertile pure bred adult Sirohi buck and in vitro culture in mCR[2]aa medium. Development of putative zygotes was observed every 24 h till day 9 post activation or fertilization under inverted phase contrast microscope. The cleavage rate, morula and blastocyst percentage in groups 1 and 2 were 67.36%, 23.07% and 9.23%, and 30.99%, 19.63% and 9.82%, respectively. The results indicated that the cleavage rate was comparatively higher following parthenogenetic activation with ionomycin/6-DMAP than IVF

Effect of different activators on development of activated in vitro matured caprine oocytes

This study was designed to compare the effectiveness of different activation treatments for activation of in vitro matured oocytes and their developmental potency in mCR[2]aa medium so as to obtain maximum number of embryos. A total of 1090 cumulus oocyte complexes [COC's] were collected from 480 ovaries. In vitro matured oocytes were randomly divided into four groups. Group 1 in vitro matured oocytes [n=226] were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP for 4 h in mCR[2]aa medium. Group 2 in vitro matured oocytes [n=294] were exposed to 7% ethanol for 5 min followed by treatment with 10 micro g/ml CHX for 4 h in mCR[2]aa medium. Group 3 in vitro matured oocytes [n=325] were exposed to 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP and 10 micro g/ml CHX for 4 h in mCR[2]aa medium. Group 4 in vitro matured oocytes [n=108] were cultured for 4 h without any chemical treatment in mCR[2]aa medium [control]. The cleavage rate in groups 1, 2, 3 and 4 was 54.42%, 44.55%, 51.69% and 0.00%, respectively. The percentage of morula and blastocyst production in group 1, group 2 and group 3 was 26.01%, 29.77% and 29.76% and 2.43%, 1.52% and 1.78%, respectively. These results suggest that the activation of in vitro matured oocytes by 7% ethanol for 5 min followed by treatment with 2.0 mM DMAP for 4 h in mCR[2]aa is most favorable for parthenogenetic caprine embryos production