本文選題：Evi-1　切入點：轉化生長因子-β超家族　出處：《山東師范大學》2011年碩士論文

【摘要】：Evi-1(the ecotropic viral integration site-1)基因作為一個逆轉錄病毒常見的插入位點,首先是在由逆轉錄病毒誘發(fā)的急性髓系白血病小鼠(AKXD)中被發(fā)現(xiàn)的。正常情況下,Evi-1基因在人的造血細胞上不表達,但在逆轉錄病毒誘導的伴染色體3q26異常的人類髓系白血病細胞上過度表達,3q26染色體也是Evi-1定位的染色體。研究發(fā)現(xiàn)Evi-1是轉化生長因子β(TGF-β)超家族信號通路中重要的調節(jié)因子,在TGF-β信號通路中發(fā)揮抑制作用。缺失Evi-1基因的小鼠將在胚胎發(fā)育的前幾周因多種生理缺陷而死亡。多項研究證實Evi-1在小鼠的胚胎組織和成體組織中均有表達。但是Evi-1在卵巢中的研究甚少。TGF-β信號通路在卵泡發(fā)育過程中的作用以及Evi-1在其通路中的抑制作用使得我們推測Evi-1在卵泡的發(fā)育過程中可能發(fā)揮某些功能。因此,本實驗的目的在于定位Evi-1在卵巢中的位置并進一步研究其在卵泡發(fā)育過程中的生物學功能。 本實驗結果顯示: 1. Evi-1蛋白定位于卵母細胞質中,在顆粒細胞,膜細胞,以及卵母細胞核中均未見其表達。 2.早期卵泡發(fā)育組,Evi-1呈現(xiàn)出高表達的狀態(tài)。在出生后1天和4天的小鼠卵巢中,Evi-1的表達量最高。隨著卵泡的發(fā)育,Evi-1在卵母細胞質中的表達量逐漸降低,在出生10天和16天的卵巢中呈現(xiàn)出一個中間級的表達量水平。最后在23天的小鼠卵巢中表現(xiàn)出一個最低量的表達。由此可見,隨著卵泡的發(fā)育,Evi-1的表達表現(xiàn)出一個逐級下降的趨勢。 3.促性腺激素處理組,Evi-1依舊表達與卵母細胞質中,并且隨著卵泡的發(fā)育其表達水平繼續(xù)呈現(xiàn)下降的趨勢。在大的有腔卵泡中,Evi-1的表達已經(jīng)非常低,但是在一些小卵泡中表現(xiàn)出高表達狀態(tài),大小卵泡的對比明顯。在PMSG24小時處理組中,大的有腔卵泡中可以看到其表達,但是在HCG4小時和HCG11小時的處理組中,大的有腔卵泡中Evi-1的表達已經(jīng)非常非常弱了。最后,在黃體中未見Evi-1的表達。 4.發(fā)情周期檢測組,Evi-1主要表達于小卵泡的卵母細胞質中。其中,發(fā)現(xiàn)Evi-1在間情期和發(fā)情期中的表達量要略高于發(fā)情前期和發(fā)情后期,提示Evi-1的表達可能受到激素的調節(jié)。 5.排卵相關基因的檢測,real-time PCR檢測Evi-1 mRNA水平和排卵相關基因(Ptgs2, Tnfaip6, Has2, Cd44, C1qbp)mRNA的水平。在HCG處理4小時后檢測發(fā)現(xiàn),Evi-1 mRNA的表達水平下調而上述排卵相關基因的水平卻上調。 總結:Evi-1在卵泡發(fā)育的早期過程中呈現(xiàn)高表達狀態(tài),隨著卵泡的發(fā)育表達水平逐漸降低。這種特定的表達方式表明Evi-1分子在早期的卵泡發(fā)育過程中可能發(fā)揮一定的作用。
[Abstract]:Evi-1(the ecotropic viral integration site-1 (Evi-1(the ecotropic viral integration site-1) gene, as a common insertion site of retrovirus, was first found in mouse acute myeloid leukemia induced by retrovirus (ARV).Under normal conditions, Evi-1 gene was not expressed in human hematopoietic cells, but overexpression of chromosome 3q26 in retrovirus-induced human myeloid leukemia cells with abnormal chromosome 3q26 was also a Evi-1 mapped chromosome.It is found that Evi-1 is an important regulatory factor in TGF- 尾) superfamily signaling pathway and plays an inhibitory role in TGF- 尾 signaling pathway.Mice that lack the Evi-1 gene will die of multiple physiological defects in the first few weeks of embryonic development.Several studies have confirmed the expression of Evi-1 in mouse embryonic and adult tissues.However, there are few studies on the role of TGF- 尾 signaling pathway in follicular development and the inhibition of Evi-1 on follicular development. It is suggested that Evi-1 may play some roles in follicular development.Therefore, the purpose of this study was to locate the position of Evi-1 in ovary and to further study its biological function during follicular development.The results show that:1.Evi-1 protein was localized in oocyte cytoplasm, but not expressed in granulosa cells, membranous cells and oocyte nuclei.2.The expression of Evi-1 in early follicular development group was high.The expression of Evi-1 was the highest in the ovaries of 1 and 4 days postnatal mice.With the development of follicle, the expression of Evi-1 in oocyte cytoplasm decreased gradually, and the expression level of Evi-1 in ovary at 10 and 16 days after birth showed an intermediate level.At last, a minimum expression was observed in the ovaries of 23 day old mice.It can be seen that the expression of Evi-1 in follicular development showed a decreasing trend.3.In gonadotropin treated group, Evi-1 was still expressed in oocyte cytoplasm, and the expression level of Evi-1 continued to decrease with the development of follicles.The expression of Evi-1 in large antral follicles has been very low, but in some small follicles, the expression of Evi-1 is high, and the contrast of large and small follicles is obvious.The expression of Evi-1 was observed in large antral follicles in PMSG24 hour group, but the expression of Evi-1 in large antral follicles was very, very weak in HCG4 hour and HCG11 hour groups.Finally, no expression of Evi-1 was found in corpus luteum.4.Evi-1 was mainly expressed in oocyte cytoplasm of small follicles in estrous cycle detection group.It was found that the expression of Evi-1 in interoestrus and estrus was slightly higher than that in preestrus and late estrus suggesting that the expression of Evi-1 might be regulated by hormones.5.The level of Evi-1 mRNA and the levels of Ptgs2, Tnfaip6, Has2, CD44, C1qbp)mRNA were detected by real-time PCR.After 4 hours of HCG treatment, it was found that the expression of Evi-1 mRNA was down-regulated and the level of ovulation-associated genes was up-regulated.It was concluded that the high expression of Evi-1 was observed in the early stage of follicular development, and the expression level decreased gradually with the development of follicle.This specific expression suggests that Evi-1 may play a role in early follicular development.
【學位授予單位】：山東師范大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R321

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本文編號：1709554