温度对人类MⅡ期卵细胞组蛋白H3K9和H4K12乙酰化状态的影响

The effect of temperature on the acetylation status of histones H3K9 and H4K12 in human MⅡoocytes

目的探讨温度对人类MⅡ期卵细胞组蛋白H3K9和H4K12乙酰化状态的影响。方法收集接受卵胞浆内单精子显微注射治疗后患者废弃的第一次减数分裂中期(MⅠ期)卵细胞体外成熟培养成的第二次减数分裂中期(MⅡ期)卵细胞,在室温下(25℃)孵育0 min、3 min、5 min和15 min后,转移至37℃复温1 h。采用间接免疫荧光染色法对MⅡ期卵细胞组蛋白H3K9和H4K12的乙酰化状态进行检测。结果与室温孵育0 min相比,孵育3 min、5 min和15 min后,检测到组蛋白H4K12的乙酰化信号的MⅡ期卵细胞比率显著增加(P<0.05)。MⅡ期卵细胞组蛋白H3K9的乙酰化信号,在室温孵育0 min,3 min、5 min和15 min后,均检测不到(P>0.05)。结论温度对MⅡ期卵细胞组蛋白H3K9的乙酰化状态无显著影响,但与组蛋白H4K12乙酰化状态密切相关。

Objective To explore the effect of temperature on the acetylation status of histones H3K9 and H4K12 in human MII oocytes.Methods The oocytes from the second meiosis metaphase(M I),which were discarded by patients after intracytoplasmic sperm injection treatment,were collected and cultured in vitro.The oocytes from the second meiosis metaphase(MⅡ)were incubated at room temperature(25℃)for 0 min,3 min,5 min and 15 min,and then transferred to 37 C for rewarming for 1 h.Indirect immunofluorescence staining was used to detect the acetylation status of histones H3K9 and H4K12 in M II oocytes.Results Compared with O min incubation at room temperature,after 3 min,5 min,and 15 min incubation,the proportion of MⅡ stage oocytes detected acetylation signals of histone H4K12 significantly increased(P<0.05).The acetylation signal of histone H3K9 in stage MⅡ oocytes was not detected after incubation at room temperature for 0 min,3 min,5 min,and 15 min(P>0.05).Conclusion Temperature has no significant effect on the acetylation state of histone H3K9 in M II stage oocytes,but is closely related to the acetylation state of histone H4K12.