Caveolae Disassembly using Methyl-β-cyclodextrin Causes the Abolition of Coupling of the Caveolae and the Sarcoplasmic Reticulum in the Rat Femoral Artery

Ashraf Albrakati
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Abstract

Background: Caveolae are essential in regulating signal transduction mechanisms of ion channels in vascular tissue, including BKCa channels (maxi-K). The current study investigated the localization of maxi-K channels within caveolae. Methods: Sixteen rats were divided into two groups: A control group and a treated group, where arteries in the treated group were incubated with methyl-β-cyclodextrin (MβCD) to disassemble caveolae from artery tissue. Immunohistochemistry (IHC), immunocytochemistry (ICC), transmission electron microscopy (TEM) and Western blot techniques were used in this study. Result: IHC of intact arteries showed colocalization of maxi-K channels with caveolin-1 in smooth muscle and endothelial cells and colocalized of maxi-K channels with caveolin-3 in smooth muscle cells only. These findings were also corroborated with ICC in a single smooth muscle cell. TEM revealed caveolae covering most plasma membranes of smooth muscle and endothelial cells and showed that caveolae sit close to the sarcoplasmic reticulum only in smooth muscle cells. TEM showed incubating arteries with MβCD led to the disassembly of caveolae from artery tissue. This study concluded that maxi-K channels localize to caveolae and that caveolae abolishment by MβCD led to the abolition of the coupling of caveolae and the sarcoplasmic reticulum.
使用甲基-β-环糊精分解腔隙导致大鼠股动脉中腔隙和肉质网耦合的消失
背景:洞穴在调节血管组织中离子通道(包括 BKCa 通道(maxi-K))的信号转导机制方面至关重要。本研究调查了 maxi-K 通道在洞穴内的定位情况。研究方法16 只大鼠分为两组:对照组和处理组,其中处理组的动脉用甲基-β-环糊精(MβCD)孵育,以分解动脉组织中的空穴。本研究采用了免疫组织化学(IHC)、免疫细胞化学(ICC)、透射电子显微镜(TEM)和 Western 印迹技术。结果完整动脉的 IHC 显示,在平滑肌细胞和内皮细胞中,maxi-K 通道与洞穴素-1 共定位;仅在平滑肌细胞中,maxi-K 通道与洞穴素-3 共定位。这些发现也在单个平滑肌细胞的 ICC 中得到了证实。TEM 显示洞穴小体覆盖了平滑肌和内皮细胞的大部分质膜,并显示只有平滑肌细胞中的洞穴小体靠近肌质网。TEM显示,用MβCD孵育动脉会导致动脉组织中的洞穴小体解体。这项研究认为,maxi-K 通道定位在洞穴体上,MβCD 取消洞穴体导致洞穴体与肌质网的耦合消失。
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