犏牛雄性不育的DNA甲基化

DNA methylation in male sterility of cattle-yak

采用Western blot法检测犏牛、牦牛睾丸组织中DNA甲基转移酶3a(Dnmt3a)蛋白的相对表达量,用甲基化定量试剂盒(荧光法,Epigentek公司)检测其全基因组DNA甲基化的水平,分析犏牛与牦牛在这2个表达量上的差异。结果显示:犏牛、牦牛睾丸组织中Dnmt3a蛋白的相对表达量分别为(0.853±0.015)和(0.651±0.016),犏牛的显著高于牦牛(P<0.05);犏牛、牦牛睾丸组织的全基因组DNA甲基化水平分别为(0.950±0.013)%和(0.808±0.016)%,犏牛的显著高于牦牛(P<0.05)。可见,犏牛睾丸组织中Dnmt3a蛋白表达水平与其全基因组DNA甲基化水平均显著高于牦牛,这可能是犏牛雄性不育的DNA甲基化机制之一,也将为犏牛雄性不育的表观遗传学研究奠定基础。

The objective of this study was to explore the possible mechanisms on DNA methylation in male sterility of cattle-yak. The expression levels of Dnmt3a protein were detected by Western blot in cattle-yak and yak testicular tissue and the levels of genome-wide DNA methylation were detected by fluorescence method using MethylFlashTM Methylated DNA Quantification Kit （Epi- gentek company） in cattle-yak and yak testicular tissue. The results showed that the relative quan- titative expression of Dnmt3a protein in cattle-yak and yak testicular tissue was （0. 853±0. 015）, （0. 651±0. 016） separately,that of cattle-yak was significantly higher than yak（P〈0.05）. The levels of genome-wide DNA methylation in cattle-yak and yak testicular tissue were （0. 950 ± 0. 013）%, （0. 808±0. 016）% separately,that in cattle-yak was significantly higher than yak （P〈 0.05）. In conclusion,the expression levels of Dnmt3a protein and genome-wide DNA methylation in cattle-yak testicular tissue were significantly higher than those in yak. It may be one of mecha- nisms on DNA methylation in male sterility of cattle-yak,which will lay the foundation for studies of epigenetics in male sterility of cattle-yak.