Characterization of multiple membrane progestin receptor (mPR) subtypes from the goldfish ovary and their roles in the induction of oocyte maturation

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• 作者：Toshinobu Tokumoto^a ; ^b ; ^c ; ^{sbttoku@ipc.shizuoka.ac.jp} ; Mika Tokumoto^a ; ^b ; Takayuki Oshima^c ; Kumi Shimizuguchi^a ; Tatsuya Fukuda^c ; Etsuko Sugita^a ; Manami Suzuki^a ; Yu-ta Sakae^a ; Yu-ichi Akiyama^a ; Ryo Nakayama^a ; Shimi Rani Roy^a ; Md. Saydur Rahman^b ; Yefei Pang^b ; Jing Dong^b ; Peter Thomas^b
• 关键词：Membrane progestin receptor ; Goldfish ; Oocyte ; Steroid binding ; Oogenesis
• 刊名：General and Comparative Endocrinology
• 出版年：2012
• 期刊代码：169_00166480
• 类别：bio
• 出版时间：15 May, 2012
• 卷：177
• 期：1
• 页码：168-176
• 文件大小：1042 K

摘要

Oocyte maturation (OM) in goldfish is induced by the maturation inducing hormone (MIH) via its membrane receptor. Previously, we described the cloning of the membrane progesterone receptor alpha (mPR伪 or paqr7b) cDNA from a goldfish ovarian cDNA library and obtained experimental evidence that the mPR伪 protein is an intermediary in MIH induction of OM in goldfish. Three mPR subtypes have been identified in fish by cDNA cloning or by in silico analysis of genome sequence databases. In order to investigate the potential roles of the mPR subtypes in oocyte maturation, we cloned additional mPRs from a goldfish ovarian cDNA library. RACE amplification, and screening of the cDNA library identified one 尾 (paqr8) and two 纬 subtypes (paqr5) (hereafter referred to as 纬-1 and 纬-2), respectively. Tissue distribution of mPR subtypes showed differential expression pattern. However, in addition to mPR伪, the 尾, 纬-1 and 纬-2 subtypes were also expressed in follicle-enclosed oocytes. Cell lines expressing the 尾, 纬-1 and 纬-2 genes were established and their steroid binding properties compared. The 尾 subtype exhibited higher binding affinity than the 纬 subtypes for 17,20尾-DHP, the MIH in goldfish. Microinjection of goldfish oocytes with a morpholino antisense oligonucleotide to mPR尾 blocked the induction of oocyte maturational competence, whereas injection of antisense oliogonucleotides to mPR纬-1 and 纬-2 were ineffective. These results suggest that the goldfish mPR尾 protein acts as an intermediary during MIH induction of OM in goldfish, in a manner similar to that described previously for mPR伪.