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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >Extraction of membrane cholesterol disrupts caveolae and impairs serotonergic (5-HT2A) and histaminergic (H1) responses in bovine airway smooth muscle: role of Rho-kinase.
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Extraction of membrane cholesterol disrupts caveolae and impairs serotonergic (5-HT2A) and histaminergic (H1) responses in bovine airway smooth muscle: role of Rho-kinase.

机译:膜胆固醇的提取破坏了牛气管并损害了牛气道平滑肌中的血清素能(5-HT2A)和组胺能(H1)反应:Rho激酶的作用。

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摘要

Some receptors and signaling molecules, such as Rho-kinase (ROCK), localize in caveolae. We asked whether the function of histamine receptors (H(1)) and 5-hydroxytryptamine (serotonin) receptors (5-HT(2A)) in bovine tracheal smooth muscle are modified after caveolae disruption and if so, whether the altered ROCK activity plays a role in this modification. Methyl-beta-cyclodextrin (MbetaCD), used to deplete membrane cholesterol, was shown to disrupt caveolae and diminish sustained contractions to histamine (approximately 80%), 5-HT (100%), alpha-methyl-5-HT (100%), and KCl (approximately 30%). Cholesterol-loaded MbetaCD (CL-MbetaCD) restored the responses to KCl and partially restored the responses to agonists. ROCK inhibition by Y-27632 diminished contractions to histamine (approximately 85%) and 5-HT (approximately 59%). 5-HT or histamine stimulation augmented ROCK activity. These increases were reduced by MbetaCD and partially reestablished by CL-MbetaCD. The increase in intracellular Ca(2+) that was induced by both agonists was reduced by MbetaCD. The presence of caveolin-1 (Cav-1), H1, 5-HT(2A), and ROCK1 was corroborated by immunoblotting of membrane fractions from sucrose gradients and by confocal microscopy. H(1) receptors coimmunoprecipitated with Cav-1 in caveolar and noncaveolar membrane fractions, whereas 5-HT(2A) receptors appeared to be restricted to noncaveolar membrane fractions. We conclude that caveolar and cholesterol integrity are indispensable for the proper functionality of the H(1) and 5-HT(2A) receptors through their Rho/ROCK signaling.
机译:一些受体和信号分子,例如Rho激酶(ROCK),位于小窝中。我们询问牛气管平滑肌破裂后是否修饰了牛气管平滑肌中的组胺受体(H(1))和5-羟色胺(5-羟色胺)(5-HT(2A))的功能,如果是,是否改变了ROCK活性在此修改中的作用。甲基-β-环糊精(MbetaCD)用于消耗膜胆固醇,已显示可破坏小孔并减少持续收缩至组胺(约80%),5-HT(100%),α-甲基-5-HT(100%) )和氯化钾(约30%)。负载胆固醇的MbetaCD(CL-MbetaCD)恢复了对KCl的反应,部分恢复了对激动剂的反应。 Y-27632对ROCK的抑制作用减少了对组胺(约85%)和5-HT(约59%)的收缩。 5-HT或组胺刺激增强了ROCK活性。这些增加被MbetaCD减少,并由CL-MbetaCD部分重新建立。 MbetaCD减少了由两种激动剂诱导的细胞内Ca(2+)的增加。小孔蛋白1(Cav-1),H1、5-HT(2A)和ROCK1的存在通过蔗糖梯度中的膜部分的免疫印迹和共聚焦显微镜得到了证实。 H(1)受体与Cav-1在小窝和非小窝膜部分中共免疫沉淀,而5-HT(2A)受体似乎仅限于非小窝膜部分。我们得出结论,对于H(1)和5-HT(2A)受体通过其Rho / ROCK信号传导的适当功能,海绵体和胆固醇的完整性是必不可少的。

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