Inoue Naoko, Manabe Noboru, Matsui Toshikatsu, Maeda Akihisa, Nakagawa Shinsuke, Wada Satoko, Miyamoto Hajime
Unit of Anatomy and Cell Biology, Department of Animal Sciences, Kyoto University, Kyoto 806-8502, Japan.
J Reprod Dev. 2003 Aug;49(4):313-21. doi: 10.1262/jrd.49.313.
To reveal the molecular mechanism of selective follicular atresia in porcine ovaries, we investigated the changes in the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptor (DR4) proteins and TRAIL mRNA in granulosa cells during follicular atresia. Immunohistochemical, Western immunoblotting and reverse transcription-polymerase chain reaction analyses (RT-PCR) revealed that significant increases in TRAIL protein and mRNA levels but not DR4 protein were changed during atresia. The RT-PCR product was confirmed to be porcine TRAIL by the cDNA sequence determination. An in vitro apoptosis inducing assay using cultured granulosa cells prepared from healthy follicles showed that TRAIL could activate caspase-3 and induce apoptotic cell death in the cells. The present findings confirm that TRAIL induces apoptosis in granulosa cells during atresia in porcine ovaries.
为揭示猪卵巢中卵泡选择性闭锁的分子机制，我们研究了卵泡闭锁过程中颗粒细胞中肿瘤坏死因子相关凋亡诱导配体（TRAIL）及其受体（DR4）蛋白表达以及TRAIL mRNA的变化。免疫组织化学、蛋白质免疫印迹和逆转录聚合酶链反应分析（RT-PCR）显示，闭锁过程中TRAIL蛋白和mRNA水平显著增加，但DR4蛋白未发生变化。通过cDNA序列测定证实RT-PCR产物为猪TRAIL。使用从健康卵泡制备的培养颗粒细胞进行的体外凋亡诱导试验表明，TRAIL可激活caspase-3并诱导细胞发生凋亡性死亡。本研究结果证实，TRAIL在猪卵巢闭锁过程中可诱导颗粒细胞凋亡。
