Oba Y, Hirai T, Yoshiura Y, Kobayashi T, Nagahama Y
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, 444-8585, Japan.
Biochem Biophys Res Commun. 2000 Sep 16;276(1):258-63. doi: 10.1006/bbrc.2000.3410.
Two thyrotropin receptor cDNAs (sTSH-Ra and sTSH-Rb) were cloned from thyroid tissue of the amago salmon, Oncorhynchus rhodurus. sTSH-Ra and sTSH-Rb showed the highest degrees of sequence homology to mammalian TSH receptors. Functional characterization in COS-7 cells transiently transfected with sTSH-Ra or sTSH-Rb showed the largest increase in cAMP when exposed to bovine TSH. RT-PCR analysis demonstrated that sTSH-Ra and sTSH-Rb were expressed in the basibranchial region, but not in the ovary, testis, liver, kidney or brain. In situ hybridization revealed that sTSH-Ra and sTSH-Rb were exclusively expressed in thyroid follicular epithelial cells of amago salmon undergoing smoltification. These results indicated that the cloned cDNAs encode functional TSH receptor proteins. This is the first report of isolation of TSH receptor molecules from nonmammalian vertebrates.
从红大麻哈鱼(Oncorhynchus rhodurus)的甲状腺组织中克隆出了两个促甲状腺激素受体cDNA(sTSH-Ra和sTSH-Rb)。sTSH-Ra和sTSH-Rb与哺乳动物促甲状腺激素受体的序列同源性最高。用sTSH-Ra或sTSH-Rb瞬时转染COS-7细胞后的功能特性表明,暴露于牛促甲状腺激素时,cAMP增加幅度最大。RT-PCR分析表明,sTSH-Ra和sTSH-Rb在鳃基部区域表达,但在卵巢、睾丸、肝脏、肾脏或大脑中不表达。原位杂交显示,sTSH-Ra和sTSH-Rb仅在正在进行降海洄游的红大麻哈鱼的甲状腺滤泡上皮细胞中表达。这些结果表明,克隆出的cDNA编码功能性促甲状腺激素受体蛋白。这是首次从非哺乳类脊椎动物中分离出促甲状腺激素受体分子的报告。