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从心肌细胞中克隆大鼠α1C - 肾上腺素能受体。α1C、α1B和α1D信使核糖核酸存在于心肌细胞中,但不存在于心脏成纤维细胞中。

Cloning of the rat alpha 1C-adrenergic receptor from cardiac myocytes. alpha 1C, alpha 1B, and alpha 1D mRNAs are present in cardiac myocytes but not in cardiac fibroblasts.

作者信息

Stewart A F, Rokosh D G, Bailey B A, Karns L R, Chang K C, Long C S, Kariya K, Simpson P C

机构信息

Division of Cardiology and Research Service, Veterans Affairs Medical Center, San Francisco, CA 94121.

出版信息

Circ Res. 1994 Oct;75(4):796-802. doi: 10.1161/01.res.75.4.796.

Abstract

alpha 1-Adrenergic receptor (AR) activation in cardiac muscle has several different physiological effects that might be mediated through different alpha 1-AR subtypes. Two alpha 1-AR subtypes have been cloned from the rat, the alpha 1B and the alpha 1D; both are present in adult rat heart. A third subtype, the alpha 1C, cloned from the cow and human, was reported to be absent in the rat. However, we recently found alpha 1C mRNA in adult rat heart by using a partial alpha 1C cDNA. Thus, all three cloned alpha 1-AR subtypes are present in the heart, but it is unknown whether each is expressed in cardiac myocytes or in cardiac fibroblasts. In the present study, the full-length rat alpha 1C-AR was cloned from cultured neonatal cardiac myocytes. alpha 1C mRNA transcripts of 3, 9.5, and 11 kb were present in adult rat heart by Northern blot analysis. alpha 1B-, alpha 1C-, and alpha 1D-subtype mRNAs were each present in isolated adult and neonatal cardiac myocytes by RNase protection assay. In addition, cultured neonatal cardiac myocytes expressed the three alpha 1-AR subtype mRNAs. In contrast, none of the alpha 1-AR mRNAs was detected in cultured neonatal cardiac fibroblasts. In addition, alpha 1-ARs were absent in fibroblasts by [3H]prazosin binding and norepinephrine-stimulated [3H]inositol phosphate production. The absence of alpha 1-ARs in cardiac fibroblasts differs from beta-adrenergic and angiotensin II receptors, which are present in both cardiac fibroblasts and cardiac myocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

心肌中的α1 - 肾上腺素能受体(AR)激活具有多种不同的生理效应,这些效应可能通过不同的α1 - AR亚型介导。已从大鼠中克隆出两种α1 - AR亚型,即α1B和α1D;两者都存在于成年大鼠心脏中。据报道,从牛和人类中克隆出的第三种亚型α1C在大鼠中不存在。然而,我们最近通过使用部分α1C cDNA在成年大鼠心脏中发现了α1C mRNA。因此,所有三种克隆的α1 - AR亚型都存在于心脏中,但尚不清楚每种亚型是否在心肌细胞或心脏成纤维细胞中表达。在本研究中,从培养的新生心肌细胞中克隆出全长大鼠α1C - AR。通过Northern印迹分析,在成年大鼠心脏中存在3、9.5和11 kb的α1C mRNA转录本。通过核糖核酸酶保护试验,α1B -、α1C - 和α1D - 亚型mRNA各自存在于分离的成年和新生心肌细胞中。此外,培养的新生心肌细胞表达三种α1 - AR亚型mRNA。相比之下,在培养的新生心脏成纤维细胞中未检测到任何α1 - AR mRNA。此外,通过[3H]哌唑嗪结合和去甲肾上腺素刺激的[3H]肌醇磷酸生成,发现成纤维细胞中不存在α1 - AR。心脏成纤维细胞中不存在α1 - AR与β - 肾上腺素能受体和血管紧张素II受体不同,后两者在心脏成纤维细胞和心肌细胞中均存在。(摘要截短于250字)

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