首页> 美国卫生研究院文献>The Journal of Clinical Investigation >Rat renal arcade segment expresses vasopressin-regulated water channel and vasopressin V2 receptor.
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Rat renal arcade segment expresses vasopressin-regulated water channel and vasopressin V2 receptor.

机译:大鼠肾弧节段表达血管加压素调节的水通道和血管加压素V2受体。

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摘要

The arcades are long, branched renal tubules which connect deep and mid-cortical nephrons to cortical collecting ducts in the renal cortex. Because they are inaccessible by standard physiological techniques, their functions are poorly understood. In this paper, we demonstrate that the arcades are a site of expression of two proteins, aquaporin-2 (the vasopressin-regulated water channel) and the V2 vasopressin receptor, that are important to regulated water transport in the kidney. Using a peptide-derived polyclonal antibody to aquaporin-2, quantitative ELISA in microdissected segments showed that aquaporin-2 is highly expressed in arcades and that the expression is increased in response to restriction of fluid intake. Immunocytochemistry revealed abundant aquaporin-2 labeling of structures in the cortical labyrinth in a pattern similar to that of the Na(+)-Ca2+ exchanger and kallikrein, marker proteins expressed in arcades but not in cortical collecting ducts. RT-PCR experiments demonstrated substantial aquaporin-2 and V2 receptor mRNA in microdissected arcades. In situ hybridization, using 35S-labeled antisense cRNA probes for the V2 receptor demonstrated strong labeling of both arcades and cortical collecting ducts. Thus, these results indicate that the arcades contain the specific proteins associated with vasopressin-regulated water transport, and may be a heretofore unrecognized site of free water absorption.
机译:拱廊是长而分支的肾小管,将深层和中层皮质肾单位连接到肾皮质中的皮质收集管。由于标准生理技术无法接近它们,因此对其功能了解甚少。在本文中,我们证明了拱廊是两个蛋白质的表达位点,aquaporin-2(加压素调节的水通道)和V2加压素受体,对调节肾脏中的水运输很重要。使用肽衍生的抗aquaporin-2的多克隆抗体,在显微切割的片段中进行定量ELISA显示,acadeporin-2在拱廊中高度表达,并且表达由于受液体摄入的限制而增加。免疫细胞化学揭示了皮质迷宫中结构丰富的aquaporin-2标记,其模式类似于Na(+)-Ca2 +交换子和激肽释放酶,激肽释放酶是在拱廊中表达的,但不在皮质收集管中表达。 RT-PCR实验表明在显微解剖的拱廊中大量存在aquaporin-2和V2受体mRNA。在原位杂交中,使用35S标记的V2受体反义cRNA探针证明了拱廊和皮质收集管的强力标记。因此,这些结果表明,所述拱廊包含与加压素调节的水运输相关的特定蛋白质,并且可能是迄今为止未认识到的自由吸水部位。

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