Cheng Yuan, Maeda Akihisa, Goto Yasufumi, Matsuda Fuko, Miyano Takashi, Inoue Naoko, Sakamaki Kazuhiro, Manabe Noboru
Animal Resource Science Center, The University of Tokyo, Kasama, Japan.
J Reprod Dev. 2008 Dec;54(6):454-9. doi: 10.1262/jrd.20088. Epub 2008 Sep 26.
Follicular selection predominantly depends on granulosa cell apoptosis in porcine ovaries, but the molecular mechanisms regulating the induction of apoptosis in granulosa cells during follicular selection remain incompletely understood. To determine the role of X-linked inhibitor of apoptosis protein (XIAP), which suppresses caspase-3, -7 and -9 activities and acts as an endogenous inhibitor of apoptotic cell death, in the regulation of granulosa cell apoptosis during follicular atresia, we examined the changes in the expression level and localization of XIAP mRNA and protein in granulosa cells during follicular atresia using reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, Western blotting and immunohistochemistry, respectively. High levels of XIAP mRNA and protein were noted in the granulosa cells of healthy follicles, and decreased levels were noted during follicular atresia. In situ hybridization and immunohistochemistry demonstrated that XIAP mRNA and protein were strongly expressed in the granulosa cells of healthy follicles, but negative/trace stainings were noted in those of atretic follicles. The present findings strongly indicate that XIAP is a candidate molecule which acts as an anti-apoptotic/pro-survival factor by inhibiting intracellular apoptosis signaling and is involved in the regulation of apoptosis in porcine granulosa cells.
在猪卵巢中,卵泡选择主要取决于颗粒细胞凋亡,但卵泡选择过程中调控颗粒细胞凋亡诱导的分子机制仍未完全清楚。为了确定X连锁凋亡抑制蛋白(XIAP)在卵泡闭锁过程中颗粒细胞凋亡调控中的作用,XIAP可抑制半胱天冬酶-3、-7和-9的活性,并作为凋亡细胞死亡的内源性抑制剂,我们分别使用逆转录-聚合酶链反应(RT-PCR)、原位杂交、蛋白质印迹和免疫组织化学技术,检测了卵泡闭锁过程中颗粒细胞中XIAP mRNA和蛋白表达水平及定位的变化。在健康卵泡的颗粒细胞中检测到高水平的XIAP mRNA和蛋白,而在卵泡闭锁过程中其水平降低。原位杂交和免疫组织化学显示,XIAP mRNA和蛋白在健康卵泡的颗粒细胞中强烈表达,但在闭锁卵泡的颗粒细胞中呈阴性/微量染色。目前的研究结果强烈表明,XIAP是一种候选分子,它通过抑制细胞内凋亡信号传导作为抗凋亡/促生存因子,并参与猪颗粒细胞凋亡的调控。