【摘要】：研究表明,在成熟前NSN構(gòu)型的卵母細胞要轉(zhuǎn)變成SN構(gòu)型,且SN型卵母細胞比NSN型卵母細胞發(fā)育能力強,但其機理尚不清楚。并且GV期卵母細胞核心組蛋白修飾變化的研究結(jié)果與染色質(zhì)構(gòu)型轉(zhuǎn)變和基因活動變化相矛盾。因此,進一步研究GV期卵母細胞染色質(zhì)構(gòu)型轉(zhuǎn)變的調(diào)控機制及其與組蛋白表觀修飾和發(fā)育能力的關(guān)系,對于了解卵母細胞成熟機理、改善成熟質(zhì)量和提高發(fā)育能力具有重要意義。我們以18-19日齡的雌性小鼠為動物模型,摘除卵巢后在體外37℃和39℃分別各保存1h和2h,使染色質(zhì)構(gòu)型與相關(guān)組蛋白修飾和發(fā)育能力進行脫鉤,進一步研究體外保存卵巢對小鼠GV期卵母細胞染色質(zhì)構(gòu)型、卵母細胞發(fā)育能力以及組蛋白表觀修飾(乙�；⒓谆�、磷酸化)的影響。研究結(jié)果表明：(1)體外保存卵巢以后卵母細胞染色質(zhì)凝集程度加重,且異常染色質(zhì)構(gòu)型增多；(2)37℃保存1h后,卵母細胞發(fā)育能力為五組中最高。相同溫度下保存,隨保存時間延長,發(fā)育能力下降；保存相同時間,溫度越高,發(fā)育能力越差；(3)同一溫度下隨保存時間延長,組蛋白(H3K14、H4K12、H4K16)乙�；健⒔M蛋白(H3K4、H3K9)甲基化水平以及組蛋白磷酸化水平(H3S10)逐漸下降,與發(fā)育能力變化趨勢一致；保存相同時間,溫度越高各種組蛋白表觀修飾水平越低,與卵母細胞發(fā)育能力變化趨勢一致；(4)隨保存溫度的升高和保存時間的延長,顆粒細胞凋亡程度加重,FasL表達水平相應(yīng)升高；(5)隨保存溫度的升高和保存時間的延長,卵母細胞Fas表達水平相應(yīng)升高。由上述結(jié)果我們可以知道體外不同條件保存卵巢以后卵母細胞染色質(zhì)凝集現(xiàn)象加重,異常染色質(zhì)構(gòu)型增多；體外不同條件保存卵巢以后卵母細胞發(fā)育能力下降,且發(fā)育能力與組蛋白表觀修飾水平趨勢一致；體外不同條件保存卵巢過程中,在顆粒細胞與卵母細胞之間Fas/FasL信號通路起到了一定的作用,Fas/FasL蛋白少量表達促進卵母細胞發(fā)育,大量表達抑制卵母細胞發(fā)育；另外,顆粒細胞輕度凋亡促進卵母細胞發(fā)育,重度凋亡抑制卵母細胞發(fā)育。
[Abstract]:The results showed that the oocytes with NSN configuration should be transformed into SN configuration before maturation, and SN oocytes had stronger developmental ability than NSN oocytes, but its mechanism was not clear. The results of GV oocyte core histone modification were inconsistent with chromatin configuration and gene activity. Therefore, further study on the regulatory mechanism of chromatin configuration transformation in GV oocytes and its relationship with histone apparent modification and developmental ability is helpful to understand the mechanism of oocyte maturation. It is important to improve maturation quality and development ability. The female mice aged 18-19 days were used as animal models. After ovariectomized, they were preserved at 37 鈩，

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