本文選題：Rac1 + 原始卵泡形成��； 參考：《中國農業(yè)大學》2016年博士論文

【摘要】：哺乳動物出生前后形成的原始卵泡庫是雌性動物一生中發(fā)育卵泡與成熟卵母細胞的唯一來源,其大小代表了雌性哺乳動物一生的生育能力。原始卵泡形成異常會導致合胞體破裂受阻、多卵子卵泡發(fā)生,從而縮短雌性動物的生育年限,降低卵母細胞的體外成熟率,最終影響生殖健康。原始卵泡的形成需要多種必要基因時空特異性地表達,但這些基因是如何協同調控的,至今仍不清楚。RhoGTPase家族的Racl分子是一個廣泛存在的信號分子開關,具有多種細胞生物學功能。研究表明,Rac1參與了許多生殖相關事件,但在原始卵泡的形成過程中是否具有功能,至今仍未見到報道。相關研究表明Rac1可以通過轉錄因子STAT3調控諸多基因的轉錄。我們推測Racl可能在原始卵泡的形成過程中具有生物學功能,并且有可能調控原始卵泡形成必要基因的轉錄。本研究發(fā)現,在小鼠原始卵泡形成過程中,Racl表達于生殖細胞中,且表達量逐步上升。為了研究Racl在小鼠原始卵泡形成過程中是否具有功能,我們利用小鼠卵巢體外培養(yǎng)體系,通過向培養(yǎng)基中添加Rac分子特異性抑制劑NSC23766和特異性敲降Racl的方法,發(fā)現抑制Racl能夠顯著抑制原始卵泡的形成,卵巢皮質區(qū)出現了大量合胞體；在胎鼠卵巢中過表達Rac1則加速原始卵泡的形成；同時,體內注射Racl抑制劑,未破裂的合胞體可以持續(xù)到青春期形成多卵子卵泡。為了探索Racl是如何調控原始卵泡形成的,我們研究了Rac1與現有的原始卵泡形成必要基因之間的關系,結果表明Racl可以調控生殖細胞中.Jagged1、GDF9、BMP15以及轉錄因子Nobox的轉錄,并且影響前體顆粒細胞中Notch2的翻譯。后續(xù)研究表明,在生殖細胞中,Rac1與STAT3直接互作促進其入核,直接激活原始卵泡形成必要基因Nobox、Jngged1、GDF9及BMP15的轉錄。受Racl調控的GDF9與BMP15可以通過旁分泌的方式作用于前體顆粒細胞,激活前體顆粒細胞中的mTORC1并進而調控Notch2的翻譯。過表達和蛋白純品添加實驗也表明Jagged1、GDF9與BMP15不僅能夠逆轉由NSC23766引起的原始卵泡形成受抑制的表型,而且添加這3種蛋白純品能夠促進原始卵泡的形成。以上結果表明：在原始卵泡的形成過程中,Racl作為關鍵性的調控因子調節(jié)了原始卵泡形成過程中必要基因的表達,從而調控了原始卵泡的形成。
[Abstract]:The primordial follicle bank formed before and after birth in mammals is the only source of follicular and matured oocytes in female animals in their lifetime, and its size represents the fertility of female mammals in their lifetime.Abnormal primordial follicle formation will lead to the rupture of syncytial cells and the formation of multiple oocytes, thus shortening the reproductive life of female animals, reducing the rate of oocyte maturation in vitro, and ultimately affecting reproductive health.The formation of primordial follicles requires a variety of necessary genes to be specifically expressed in time and space, but how these genes are co-regulated remains unclear. The Racl molecule of the RhoGTPase family is a widespread signal molecule switch.It has many cell biological functions.Studies have shown that Rac1 is involved in many reproductive events, but whether it has function in the formation of primordial follicles has not been reported.Related studies have shown that Rac1 can regulate the transcription of many genes through the transcription factor STAT3.We speculate that Racl may have biological function in the process of primordial follicle formation and may regulate the transcription of genes necessary for primordial follicle formation.In this study, it was found that Racl was expressed in germ cells during primordial follicle formation in mice, and the expression level increased gradually.In order to study the function of Racl in the process of mouse primordial follicle formation, we used the mouse ovary culture system in vitro by adding Rac molecular specific inhibitor NSC23766 and specific knockout Racl to the culture medium.It was found that inhibition of Racl could significantly inhibit the formation of primordial follicles, and a large number of syncytial bodies appeared in the ovarian cortex; overexpression of Rac1 in fetal ovaries accelerated the formation of primordial follicles; at the same time, Racl inhibitor was injected in vivo.Unruptured syncytial can continue to form multiple oocyte follicles until puberty.In order to explore how Racl regulates primordial follicle formation, we studied the relationship between Rac1 and the necessary genes for primordial follicle formation. The results showed that Racl could regulate the transcription of. Jagged1, GDF9, BMP15 and Nobox.It also affects the translation of Notch2 in precursor granulosa cells.Further studies have shown that the direct interaction between rac1 and STAT3 in germ cells promotes the nucleation of the primordial follicles and activates the transcription of GDF9 and BMP15, which are essential genes for the formation of primordial follicles.GDF9 and BMP15 regulated by Racl can act on precursor granulosa cells through paracrine way, activate mTORC1 in precursor granulosa cells and then regulate the translation of Notch2.Overexpression and protein addition also showed that Jagged1GDF9 and BMP15 could not only reverse the inhibited phenotype of primordial follicles induced by NSC23766, but also promote the formation of primordial follicles.These results suggest that Racl, as a key regulatory factor, regulates the expression of necessary genes in the process of primordial follicle formation and thus regulates the formation of primordial follicles.
【學位授予單位】：中國農業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：Q492

【相似文獻】

相關期刊論文 前7條

1 吳磊,鄭莉萍,鄭月慧;原始卵泡活化的相關研究進展[J];生殖醫(yī)學雜志;2005年05期

2 高輝;郁琦;徐苓;;原始卵泡的形成與發(fā)育[J];生殖醫(yī)學雜志;2007年06期

3 郭淑華;;相關分子在哺乳動物原始卵泡啟動和生長中的作用[J];濰坊學院學報;2010年06期

4 徐良全;許愛霞;黃健;陳蔚云;鄭月慧;;c-erbB_2在大鼠卵巢中的表達及其在原始卵泡生長啟動中的作用[J];生理學報;2009年05期

5 柳海珍,劉以訓;EGF在新生大鼠原始卵泡生長啟動中的作用[J];中國科學C輯:生命科學;2000年03期

6 孫理蘭;谷朝勇;潘慶杰;沈偉;;原始卵泡形成與早期卵泡發(fā)育的遺傳調控[J];生物技術通訊;2008年01期

7 ;[J];;年期

相關會議論文 前6條

1 雷蕾;周波;楊R，

本文編號：1764507