Characterization of Gastric Na+/I−Symporter of the Rat

Tomio Kotani , Yoshikazu Ogata , Ikuo Yamamoto , Yatsuki Aratake , Jun-Ichi Kawano , Tatsuo Suganuma , Sachiya Ohtaki
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引用次数: 50

Abstract

Characterization of gastric Na+/Isymporter (NIS) of the rat was carried out. Sequencing of the open reading frame of gastric NIS mRNA showed only three nucleotide changes when compared with FRTL-5 NIS cDNA, and two of these changes led to amino acid changes. The results of Northern blot analysis showed that abundant NIS mRNA was expressed in the stomach when compared with other organs. Western blot analysis using gastric mucosa and FRTL-5 lysates detected the difference in molecular weight between FRTL-5 and gastric mucosa lysates, suggesting abnormal posttranslational modification of gastric NIS protein. Immunohistochemically, gastric NIS protein was located in the cornification layer of the stratified squamous epithelium of the pars proventricularis and in parietal cells and on the apical border of surface epithelial cells of the pars glandularis. Gastric NIS protein was present in tubulovesicular structures and lysosomes in parietal cells by immunoelectron microscopy. Gastric NIS protein exists to trap Ifrom the gastric lumen, except in parietal cells. Results indicated that a very large amount of gastric NIS mRNA is expressed to be translated, whereas only a small amount of immature gastric NIS protein is detected. This may indicate that immature gastric NIS protein rapidly degrades to peptides.

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大鼠胃Na+/I−同向转运体的表征
对大鼠胃Na+/I−同调体(NIS)进行表征。与FRTL-5 NIS cDNA相比,胃NIS mRNA开放阅读框测序显示只有3个核苷酸变化,其中两个变化导致氨基酸变化。Northern blot分析结果显示,与其他器官相比,胃中表达了丰富的NIS mRNA。利用胃黏膜和FRTL-5裂解物进行Western blot分析,检测到FRTL-5和胃黏膜裂解物的分子量差异,提示胃NIS蛋白翻译后修饰异常。免疫组化结果显示,胃NIS蛋白位于脑室前部分层鳞状上皮的锥化层、腺部表面上皮细胞的顶细胞和顶缘。免疫电镜观察发现胃NIS蛋白存在于胃壁细胞的管泡结构和溶酶体中。除了胃壁细胞外,胃NIS蛋白的存在是为了从胃管中捕获I -。结果表明,大量的胃NIS mRNA被表达翻译,而只有少量未成熟的胃NIS蛋白被检测到。这可能表明未成熟的胃NIS蛋白迅速降解为多肽。
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