Terao Yasuko, Kumano Satoshi, Takatsu Yoshihiro, Hattori Masahiko, Nishimura Atsushi, Ohtaki Tetsuya, Shintani Yasushi
Pharmacology Research Laboratories I, Pharmaceutical Research Division, Takeda Chemical Industries, Ltd., 17-85 Jusohonmachi 2-chome, Yodogawa, Osaka 532-8686, Japan.
Biochim Biophys Acta. 2004 May 25;1678(2-3):102-10. doi: 10.1016/j.bbaexp.2004.02.005.
Metastin is encoded by a putative human metastasis suppressor gene KiSS-1, and is the cognate ligand of a G-protein-coupled receptor designated OT7T175. To study the physiological function(s) of metastin, we cloned rat and mouse KiSS-1 cDNAs both encoding 130-amino acid KiSS-1 proteins. Sequence analysis suggested that processing of the rat and mouse KiSS-1 proteins produces 52-amino-acid peptides, each with an amidated carboxyl terminal and with a single possible disulfide bond, corresponding to rat and mouse metastins. The carboxyl-terminal sequence of metastin, known to be essential for functional receptor interaction, was found to be highly conserved among humans and rodents. Real-time PCR analysis indicated that rat KiSS-1 mRNA showed the highest expression level in the cecum and colon. Since KiSS-1 mRNA and metastin are known to be abundant in human placenta, we further studied the localization of KiSS-1 and OT7T175 mRNAs in rat placenta by in situ hybridization. KiSS-1 and OT7T175 mRNAs were specifically detected in trophoblast giant cells at embryonic day 12.5, and the transcripts in the cells gradually decreased during placental maturation. These results suggest that metastin/OT7T175 signaling may participate in implantation of the mammalian embryo, placenta formation, and maintenance of pregnancy.
转移抑制素由一个假定的人类转移抑制基因KiSS-1编码,是一种名为OT7T175的G蛋白偶联受体的同源配体。为了研究转移抑制素的生理功能,我们克隆了大鼠和小鼠的KiSS-1 cDNA,二者均编码130个氨基酸的KiSS-1蛋白。序列分析表明,大鼠和小鼠的KiSS-1蛋白加工后产生52个氨基酸的肽段,每个肽段的羧基末端酰胺化,且只有一个可能的二硫键,这与大鼠和小鼠的转移抑制素相对应。转移抑制素的羧基末端序列已知对于功能性受体相互作用至关重要,在人类和啮齿动物中高度保守。实时PCR分析表明,大鼠KiSS-1 mRNA在盲肠和结肠中表达水平最高。由于已知KiSS-1 mRNA和转移抑制素在人胎盘中含量丰富,我们通过原位杂交进一步研究了大鼠胎盘内KiSS-1和OT7T175 mRNA的定位。在胚胎第12.5天,在滋养层巨细胞中特异性检测到KiSS-1和OT7T175 mRNA,随着胎盘成熟,这些细胞中的转录本逐渐减少。这些结果表明,转移抑制素/OT7T175信号通路可能参与哺乳动物胚胎的着床、胎盘形成和妊娠维持。