Goto Yasufumi, Matsuda-Minehata Fuko, Inoue Naoko, Matsui Toshikatsu, Maeda Akihisa, Manabe Noboru
Unit of Anatomy and Cell Biology, Department of Animal Sciences, Kyoto University, Japan.
J Reprod Dev. 2004 Oct;50(5):549-55. doi: 10.1262/jrd.50.549.
To reveal the molecular regulation mechanism of selective follicular atresia in porcine ovaries, we isolated the porcine cDNA encoding cellular FLICE-like inhibitory protein (cFLIP), which inhibits death receptor-mediated apoptosis signal transduction. Two alternative splicing isoforms of cFLIP, porcine cellular FLIP-short form (pcFLIPS, 642 bp and 214-aa) and -long form (pcFLIPL, 1446 bp and 482-aa), were identified from a cDNA library prepared from follicular granulosa cells of pig ovaries. pcFLIPS and pcFLIPL indicated high identities with human and murine cFLIP, and both of them contain two tandem specific amino acid regions (death effector domain: DED) in their N-terminal, suggesting that pcFLIPS and pcFLIPL inhibit the death receptor-mediated apoptosis signal by binding to other pro-apoptotic factors mediated by DED. pcFLIPS contains a short C-terminal region, while pcFLIPL has a caspase-like domain in the C-terminal region. The reverse transcription-polymerase chain reaction analysis revealed that both pcFLIPS and pcFLIPL mRNAs were highly expressed in granulosa cells of healthy follicles, suggesting that these cFLIPs play important roles in the regulation mechanism of apoptosis in ovarian follicular granulosa cells. The present data will contribute to understanding of the physiological roles of cFLIPs in the apoptosis regulation in porcine tissues.
为揭示猪卵巢中选择性卵泡闭锁的分子调控机制，我们分离出了编码细胞FLICE样抑制蛋白（cFLIP）的猪cDNA，该蛋白可抑制死亡受体介导的凋亡信号转导。从猪卵巢卵泡颗粒细胞制备的cDNA文库中鉴定出了cFLIP的两种可变剪接异构体，即猪细胞FLIP短型（pcFLIPS，642 bp和214个氨基酸）和长型（pcFLIPL，1446 bp和482个氨基酸）。pcFLIPS和pcFLIPL与人及小鼠的cFLIP具有高度同源性，且二者在其N端均含有两个串联的特定氨基酸区域（死亡效应结构域：DED），这表明pcFLIPS和pcFLIPL通过与DED介导的其他促凋亡因子结合来抑制死亡受体介导的凋亡信号。pcFLIPS含有一个短的C端区域，而pcFLIPL在C端区域具有一个半胱天冬酶样结构域。逆转录-聚合酶链反应分析显示，pcFLIPS和pcFLIPL的mRNA在健康卵泡的颗粒细胞中均高表达，这表明这些cFLIPs在卵巢卵泡颗粒细胞凋亡调控机制中发挥重要作用。本研究数据将有助于理解cFLIPs在猪组织凋亡调控中的生理作用。
