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血管紧张素II 1型受体亚型mRNA在大鼠肾脏中的定位

Localization of angiotensin II type 1 receptor subtype mRNA in rat kidney.

作者信息

Healy D P, Ye M Q, Troyanovskaya M

机构信息

Department of Pharmacology, Mount Sinai School of Medicine, City University of New York, New York 10029.

出版信息

Am J Physiol. 1995 Feb;268(2 Pt 2):F220-6. doi: 10.1152/ajprenal.1995.268.2.F220.

Abstract

The physiological effects of angiotensin II (ANG II) on the kidney are mediated primarily by the ANG II type 1 (AT1) receptor. Two highly similar AT1 receptor subtypes have been identified in the rat by molecular cloning techniques, namely AT1A and AT1B. The intrarenal localization of the AT1A and AT1B receptor subtypes has not been studied by hybridization methods with subtype-specific receptor probes. Using radiolabeled probes from the 3' noncoding region of the AT1A and AT1B cDNAs, we localized AT1 mRNA in rat kidney by in situ hybridization. Specificity of the 3' noncoding region probes was tested by Northern blot and solution hybridization methods. AT1A mRNA levels were highest in the liver, kidney, and adrenal. In contrast, AT1B mRNA levels were highest in the adrenal and pituitary and low in kidney. Autoradiographic localization of 125I-[Sar1,Ile8]ANG II binding indicated that the highest levels of AT1 receptors were found in glomeruli and vascular elements. In situ hybridization with a nonselective AT1 receptor riboprobe indicated that the highest levels of AT1 mRNA were in the outer medullary vasa recta and cortical glomeruli with additional diffuse labeling of the cortex and outer medulla, consistent with labeling of tubular elements. In contrast, in situ hybridization with the AT1 subtype selective probes revealed that AT1A receptor mRNA was primarily localized to the vasa recta and diffusely to the outer stripe of the outer medulla and the renal cortex.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

血管紧张素II(ANG II)对肾脏的生理作用主要由1型血管紧张素II(AT1)受体介导。通过分子克隆技术在大鼠中已鉴定出两种高度相似的AT1受体亚型,即AT1A和AT1B。尚未通过使用亚型特异性受体探针的杂交方法研究AT1A和AT1B受体亚型在肾内的定位。我们使用来自AT1A和AT1B cDNA 3'非编码区的放射性标记探针,通过原位杂交在大鼠肾脏中定位AT1 mRNA。通过Northern印迹和溶液杂交方法测试了3'非编码区探针的特异性。AT1A mRNA水平在肝脏、肾脏和肾上腺中最高。相比之下,AT1B mRNA水平在肾上腺和垂体中最高,而在肾脏中较低。125I-[Sar1,Ile8]ANG II结合的放射自显影定位表明,AT1受体的最高水平存在于肾小球和血管成分中。用非选择性AT1受体核糖探针进行原位杂交表明,AT1 mRNA的最高水平位于外髓质直小血管和皮质肾小球,皮质和外髓质有额外的弥漫性标记,这与肾小管成分的标记一致。相比之下,用AT1亚型选择性探针进行原位杂交显示,AT1A受体mRNA主要定位于直小血管,并弥漫性定位于外髓质的外带和肾皮质。(摘要截短于250字)

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