Identification of the mRNA for the novel alpha 1D-adrenoceptor and two other alpha 1-adrenoceptors in vascular smooth muscle.
Identification of the mRNA for the novel alpha 1D-adrenoceptor and two other alpha 1-adrenoceptors in vascular smooth muscle.
复制标题
DOI：
--
复制
发表时间：
1994-07
影响因子：
3.6
通讯作者：
M. Piascik;M. S. Smith;E. E. Soltis-E.;D. Perez
中科院分区：
文献类型：
--
作者：
M. Piascik;M. S. Smith;E. E. Soltis-E.;D. Perez
原位杂交组织化学，放射性配体结合，并在体外收缩的研究被用来表征最近发现的α 1D-肾上腺素受体的血管分布。原位杂交与反义探针定位的α 1D-肾上腺素受体的mRNA的大鼠主动脉，肾和肠系膜阻力动脉的中层。如果组织首先用RNase处理或使用有义探针，则未检测到特异性杂交信号。该受体表达为蛋白质的程度用放射性配体结合研究进行评估。一系列用于表征α 1-肾上腺素受体的配体与主动脉和肠系膜血管床匀浆中[3 H]哌唑嗪标记的两个位点相互作用。高亲和力位点具有α 1A-肾上腺素受体的特征。然而，低亲和力位点的配体结合特性不同于α 1 D-肾上腺素受体或任何其他已知的α 1-肾上腺素受体亚型。在主动脉、肾动脉和肠系膜阻力动脉中也检测到α 1B和α 1C肾上腺素受体的mRNA。氯乙基可乐定（CEC）（1和10 μ M）对苯肾上腺素诱导的血管平滑肌收缩有不同的影响。CEC完全抑制主动脉的反应，并引起肠系膜阻力动脉的部分抑制。相同浓度的CEC对肾动脉苯肾上腺素反应的影响不大。数据表明如下。1)新型α 1D肾上腺素受体的mRNA定位于血管平滑肌。2)这种受体的表达的连续鉴定是不可能的;这可能与α 1-肾上腺素受体的其他亚型的共表达有关。3)在大鼠外周血管中检测到α 1C-肾上腺素受体的mRNA。4)α 1A-肾上腺素受体也位于脉管系统中。5)三个和可能四个α 1-肾上腺素受体参与血管平滑肌调节。
In situ hybridization histochemistry, radioligand binding, and in vitro contractile studies were used to characterize the vascular distribution of the recently discovered alpha 1D-adrenoceptor. In situ hybridization with an antisense probe localized the mRNA for the alpha 1D-adrenoceptor to the medial layer of the rat aorta renal and mesenteric resistance arteries. If the tissues were first treated with RNase or a sense probe was used, no specific hybridization signal was detected. The extent to which this receptor was expressed as protein was assessed with radioligand binding studies. A series of ligands used to characterize alpha 1-adrenoceptors interacted with two sites labeled by [3H]prazosin in homogenates from the aorta and mesenteric vascular bed. The high affinity site had the characteristics of an alpha 1A-adrenoceptor. However, the low affinity site had ligand binding characteristics distinct from those of the alpha 1D-adrenoceptor or any other known alpha 1-adrenoceptor subtype. mRNA for the alpha 1B- and alpha 1C-adrenoceptors was also detected in the aorta, renal arteries, and mesenteric resistance arteries. Chloroethylclonidine (CEC) (1 and 10 microM) had differential effects on phenylephrine-induced contractions of vascular smooth muscle. CEC completely inhibited the response in the aorta and caused a partial inhibition in the mesenteric resistance artery. The same concentrations of CEC had little effect on phenylephrine responses in the renal artery. The data suggest the following. 1) mRNA for the novel alpha 1D-adrenoceptor is localized in vascular smooth muscle. 2) Definitive identification of expression of this receptor was not possible; this may be related to coexpression of other subtypes of alpha 1-adrenoceptors. 3) mRNA for the alpha 1C-adrenoceptor was detected in rat peripheral vasculature. 4) The alpha 1A-adrenoceptor is also localized in the vasculature. 5) Three and possibly four alpha 1-adrenoceptors participate in vascular smooth muscle regulation.