牛卵母细胞减数分裂期组蛋白H4K12乙酰化的变化

Acetylization Change of Histone H4K12 in Ovocyte of Cattle during Reduction Division Phase

试验以牛卵母细胞作为材料,以曲古抑菌素A作为去乙酰化酶(HDAC)抑制剂。应用激光共聚焦显微镜技术,通过检测卵母细胞组蛋白H4K12乙酰强度,探讨乙酰基转移酶(HAT)和组蛋白去乙酰基酶(HDAC)在减数分裂Ⅰ、Ⅱ期的活性。TSA的处理浓度为80μmol/L。结果显示,组蛋白H4K12乙酰荧光强度在卵母细胞减数分裂期完全消失;在TSA存在下培养成熟的卵母细胞,在减数分裂期检测到强的组蛋白H4K12乙酰化荧光强度;然后卵母细胞移入不含TSA的培养液中,3h后组蛋白H4K12乙酰荧光强度再次完全消失。结论:HAT在减数分裂Ⅰ、Ⅱ期无活性,HDAC在减数分裂Ⅰ、Ⅱ期有活性。

In the trial,ovocyte of cattle was used as material and trichostatin-A was used as suppressor of de-acetylase.The intensity of Acetylization of histone H4K12 was determined by laser confocal microscopy technique to explore the activity of transacetylase（HAT） and histon de-acylase（HDAC） during reduction division phase of Ⅰand Ⅱ,the treatment concentration of TSA was 80 μmol/L.The result indicated that fluorescence intensity of histone H4K12 couldn＇t be detected during reduction division phase,and it could be detected from matured ovocytes under the culture condition of TSA,but histone H4K12 disappear completely after the ovocytes were transferred into free-TSA culture medium for 3 hour later.The conclusion could be made that there isn＇t activity to HAT,but HDAC was activity during reduction division phase of Ⅰand Ⅱ.