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大鼠肾内血管紧张素II 1型受体mRNA的定位

Intrarenal localization of angiotensin II type 1 receptor mRNA in the rat.

作者信息

Kakinuma Y, Fogo A, Inagami T, Ichikawa I

机构信息

Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee.

出版信息

Kidney Int. 1993 Jun;43(6):1229-35. doi: 10.1038/ki.1993.174.

Abstract

We examined intrarenal localization of angiotensin II type 1 receptor (AT1) mRNA in kidneys of normal adult male Munich Wistar rats using the methods of reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. For RT-PCR, we used a rat AT1 subtype A (AT1A)-specific oligonucleotide primer pair. To semi-quantitatively assess the expression level of AT1 mRNA among several regions of kidney, AT1 cDNA was coamplified with beta-actin cDNA. When compared to the level in the adrenal gland (expressed as 100%), the level of AT1 mRNA was markedly higher in glomeruli (273 +/- 69%), followed in intensity by the renal papilla (151 +/- 57%), renal cortex (139 +/- 19%), and renal medulla (114 +/- 35%). In situ hybridization studies, using a 479 bp nucleotide fragment from AT1A-coding exon as a probe, also revealed a glomerular preponderant pattern of AT1 mRNA localization. Thus, within the glomerulus, AT1 mRNA localized in mesangial areas, predominantly at the vascular pole. In the vascular components of the juxtaglomerular apparatus (JGA), namely the terminal portion of the afferent arteriole (that is, immunohistochemically renin-positive site) and extraglomerular mesangial cells, the latter showed AT1 mRNA localization in the non-manipulated kidney, while AT1 mRNA was undetectable in the arteriole outside the JGA. The kidneys of rats treated with an angiotensin I converting enzyme inhibitor (ACEI) showed extension of the AT1 mRNA localization on the afferent arteriole toward the interlobular artery.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

我们采用逆转录-聚合酶链反应(RT-PCR)和原位杂交方法,研究了正常成年雄性慕尼黑Wistar大鼠肾脏中血管紧张素II 1型受体(AT1)mRNA的肾内定位。对于RT-PCR,我们使用了大鼠AT1 A亚型(AT1A)特异性寡核苷酸引物对。为了半定量评估肾脏几个区域中AT1 mRNA的表达水平,将AT1 cDNA与β-肌动蛋白cDNA共同扩增。与肾上腺中的水平(表示为100%)相比,肾小球中AT1 mRNA的水平明显更高(273±69%),其次是肾乳头(151±57%)、肾皮质(139±19%)和肾髓质(114±35%)。原位杂交研究使用来自AT1A编码外显子的479 bp核苷酸片段作为探针,也揭示了AT1 mRNA定位的肾小球优势模式。因此,在肾小球内,AT1 mRNA定位于系膜区域,主要在血管极。在肾小球旁器(JGA)的血管成分中,即入球小动脉的末端部分(即免疫组织化学上肾素阳性位点)和球外系膜细胞,在未处理的肾脏中,后者显示有AT1 mRNA定位,而在JGA外的小动脉中未检测到AT1 mRNA。用血管紧张素I转换酶抑制剂(ACEI)处理的大鼠肾脏显示,入球小动脉上的AT1 mRNA定位向小叶间动脉延伸。(摘要截断于250字)

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