两种培养液对牦牛孤雌激活胚胎发育及DNMT1基因表达的影响

Effects of two culture media on the development of yak embryos and expression of DNMT1 gene after parthenogenetic activation

为寻求较佳的牦牛胚胎培养液,将成熟的牦牛卵母细胞进行孤雌激活后分别置于培养液SOFaa和G1/G2中进行发育培养,观察培养开始后第24、48、96、120和168小时胚胎的发育情况,并采用实时荧光定量PCR检测DNMT1基因的表达量。结果显示,SOFaa液中2-4细胞胚胎、4-8细胞胚胎、9-16细胞胚胎、桑葚胚和囊胚的发育率分别为87.50%、68.00%、40.00%、34.29%、12.86%;而G1/G2联合培养液中2-4细胞胚胎、4-8细胞胚胎、9-16细胞胚胎、桑葚胚和囊胚的发育率分别为89.33%、81.33%、58.67%、49.33%、17.14%。胚胎中DNMT1基因在G1/G2联合培养液中的表达量显著高于SOFaa液中的表达量。以上结果表明,G1/G2联合培养液可通过调控DNMT1基因的表达而提高胚胎发育率,可作为牦牛胚胎体外培养的较佳培养液。

In order to explore a better condition for culturing yak embryos,yak oocytes in SOFaa and G1/G2 were cultured after parthenogenetic activation.The embryo development rate at hour 24,48,96,120 and 168post-starting of culturing was analyzed,and the expression of DNMT1 gene was detected by the real-time RCR.In result,the development rate of 2-4cell embryo,4-8cell embryo,9-16 cell embryo,morula and blastocyst in the SOFaa medium were 87.50%,68.00%,40.00%,34.29% and 12.86%,respectively,while in the G1/G2 coculture condition the development rate were 89.33%,81.33%,58.67%,49.33%and17.14%,respectively.The expression quantity of DNMT1 gene in the embryos in the G1/G2 coculture medium was higher than that in the SOFaa fluid.The above-mentioned results showed that G1/G2 coculture medium can increase embryonic development rate by regulating the expression of DNMT1 gene,which can be used as a better in vitro medium for yak embryos.