發(fā)布時間：2019-07-10 15:16

【摘要】：目的:探討自噬相關蛋白微管相關蛋白1輕鏈3(microtubule associated protein 1 light chain 3,LC3)在小鼠卵泡顆粒細胞中的表達及定位。方法:昆明小鼠腹腔注射孕馬血清促性腺激素(pregnant mare serum gonadotropin,PMSG)前及注射后的第1、2、3、4、5天,采用免疫組織化學染色方法檢測自噬相關蛋白LC3和凋亡相關蛋白cleaved caspase-3在卵泡中表達及共定位情況;促卵泡激素(follicle stimulating hormone,FSH)處理顆粒細胞后,Western blot檢測LC3和cleaved caspase-3在顆粒細胞中的水平,免疫熒光方法對LC3在顆粒細胞中的亞細胞定位進行研究。結果:在小鼠卵泡發(fā)育的各個階段,LC3蛋白主要在顆粒細胞中表達;免疫熒光顯示在閉鎖卵泡的顆粒細胞中cleaved caspase-3和LC3的蛋白水平較高;在PMSG腹腔注射后第1、2天,顆粒細胞cleaved caspase-3和LC3-II的蛋白水平減少(P0.05);在PMSG腹腔注射后第3、4、5天,顆粒細胞的cleaved caspase-3和LC3-II蛋白水平依次增加;LC3-II和cleaved caspase-3的蛋白水平具有相關性(r~2=0.8299,P0.05)。FSH處理后,LC3-II定位于顆粒細胞胞漿并呈點狀分布。結論:LC3主要在卵泡顆粒細胞中表達,其表達具有細胞特異性和區(qū)域特異性,提示自噬在卵泡發(fā)育過程中主要在顆粒細胞中發(fā)生。卵巢顆粒細胞自噬和凋亡具有相關性,均呈促性腺激素依賴性。
文內(nèi)圖片：
圖片說明：小鼠卵巢不同發(fā)育階段的卵泡中LC3定位表達情況
[Abstract]:Aim: to investigate the expression and localization of autophagy associated protein microtubule associated protein 1 light chain 3 (microtubule associated protein 1 light chain 3 in mouse follicular granulosa cells. Methods: the expression and co-localization of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 in follicles were detected by immunohistochemical staining before and on the 1st, 2nd, 3rd and 4th day after intraabdominal injection of serum gonadotropin cleaved caspase-3 in Kunming mice. The levels of LC3 and cleaved caspase-3 in granulosa cells were detected by, Western blot after treatment with follicular hormone (follicle stimulating hormone,FSH). The subcellular localization of LC3 in granulosa cells was studied by immunofluorescence. Results: at all stages of follicular development in mice, LC3 protein was mainly expressed in granulosa cells, immunofluorescence showed that the protein levels of cleaved caspase-3 and LC3 in granulosa cells of atretic follicles were higher, the protein levels of cleaved caspase-3 and LC3-II in granulosa cells decreased on the 1st and 2nd day after intraabdominal injection of PMSG (P 0.05), and the levels of cleaved caspase-3 and LC3-II protein in granulosa cells increased in turn on the 3rd and 4th day after intraabdominal injection of PMSG. There was a correlation between the protein level of LC3-II and cleaved caspase-3 (r 鈮，

本文編號：2512683