LC3蛋白在小鼠卵泡颗粒细胞中的表达及定位研究
发布时间：2019-07-10 15:16
【摘要】：目的:探讨自噬相关蛋白微管相关蛋白1轻链3(microtubule associated protein 1 light chain 3,LC3)在小鼠卵泡颗粒细胞中的表达及定位。方法:昆明小鼠腹腔注射孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)前及注射后的第1、2、3、4、5天,采用免疫组织化学染色方法检测自噬相关蛋白LC3和凋亡相关蛋白cleaved caspase-3在卵泡中表达及共定位情况;促卵泡激素(follicle stimulating hormone,FSH)处理颗粒细胞后,Western blot检测LC3和cleaved caspase-3在颗粒细胞中的水平,免疫荧光方法对LC3在颗粒细胞中的亚细胞定位进行研究。结果:在小鼠卵泡发育的各个阶段,LC3蛋白主要在颗粒细胞中表达;免疫荧光显示在闭锁卵泡的颗粒细胞中cleaved caspase-3和LC3的蛋白水平较高;在PMSG腹腔注射后第1、2天,颗粒细胞cleaved caspase-3和LC3-II的蛋白水平减少(P0.05);在PMSG腹腔注射后第3、4、5天,颗粒细胞的cleaved caspase-3和LC3-II蛋白水平依次增加;LC3-II和cleaved caspase-3的蛋白水平具有相关性(r~2=0.8299,P0.05)。FSH处理后,LC3-II定位于颗粒细胞胞浆并呈点状分布。结论:LC3主要在卵泡颗粒细胞中表达,其表达具有细胞特异性和区域特异性,提示自噬在卵泡发育过程中主要在颗粒细胞中发生。卵巢颗粒细胞自噬和凋亡具有相关性,均呈促性腺激素依赖性。
文内图片：
图片说明：小鼠卵巢不同发育阶段的卵泡中LC3定位表达情况
[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，
本文编号：2512683
文内图片：
图片说明：小鼠卵巢不同发育阶段的卵泡中LC3定位表达情况
[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，
本文编号：2512683
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