Mistry Abinash Chandra, Chen Guangping, Kato Akira, Nag Kakon, Sands Jeff M, Hirose Shigehisa
Dept. of Biological Sciences, Tokyo Institute of Technology, 4259-B-19 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Am J Physiol Renal Physiol. 2005 Mar;288(3):F455-65. doi: 10.1152/ajprenal.00296.2004. Epub 2004 Sep 21.
A new type of urea transporter was identified by a database search and shown to be highly expressed in the renal proximal tubule cells of teleosts; proximal tubule-type urea transporters have not been describe previously. We first identified urea transporter-like sequences in the fugu genome and in an EST database of rainbow trout. Based on these pieces of sequence information, we obtained a full-length cDNA for the eel ortholog, consisting of 378 amino acid residues, and named it eUT-C. Although its sequence similarity to the known urea transporters is low (approximately 35%), its heterologous expression in Xenopus laevis oocytes indicated that it is a facilitative urea transporter sensitive to phloretin. Its activity is not dependent on Na+. Northern blot analysis showed that expression of eUT-C is highly restricted to the kidney, with weak expression in the stomach. In both tissues, eUT-C mRNA was strongly induced when eels were transferred from freshwater to seawater. Immunohistochemistry and in situ hybridization histochemistry revealed proximal tubule cell localization of eUT-C. Taking into account that 1) urea is mainly secreted from the gill where another type of urea transporter (eUT) has been identified and 2) fish excrete a very small volume of urine in seawater, we propose that eUT-C cloned here is a key component working in combination with the gill transporter to achieve an efficient urea excretory system in fish, namely, eUT-C reabsorbs urea from glomerular filtrate and sends it to the gill, through the circulation, for excretion.
通过数据库搜索鉴定出一种新型尿素转运蛋白,结果表明其在硬骨鱼的肾近端小管细胞中高度表达;此前尚未描述过近端小管型尿素转运蛋白。我们首先在河豚基因组和虹鳟鱼的一个EST数据库中鉴定出尿素转运蛋白样序列。基于这些序列信息,我们获得了鳗鱼直系同源物的全长cDNA,其由378个氨基酸残基组成,并将其命名为eUT-C。尽管其与已知尿素转运蛋白的序列相似性较低(约35%),但其在非洲爪蟾卵母细胞中的异源表达表明它是一种对根皮素敏感的易化型尿素转运蛋白。其活性不依赖于Na+。Northern印迹分析表明,eUT-C的表达高度局限于肾脏,在胃中表达较弱。在这两种组织中,当鳗鱼从淡水转移到海水时,eUT-C mRNA均被强烈诱导。免疫组织化学和原位杂交组织化学显示eUT-C定位于近端小管细胞。考虑到1)尿素主要从已鉴定出另一种尿素转运蛋白(eUT)的鳃分泌,以及2)鱼类在海水中排出的尿量非常少,我们提出这里克隆的eUT-C是与鳃转运蛋白协同工作以实现鱼类高效尿素排泄系统的关键成分,即eUT-C从肾小球滤液中重吸收尿素,并通过循环将其输送到鳃进行排泄。