Recording of Inward Rectifying K+ Currents in Freshly Isolated Basilar Artery Smooth Muscle Cells by Patch Clamp Technique.

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES Jove-Journal of Visualized Experiments Pub Date : 2025-02-07 DOI:10.3791/67414
Pengmei Guo, Weiping Li, Wenqiao An
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Abstract

Cerebrovascular disease is a prevalent condition among the elderly, with its incidence steadily rising. The basilar artery is a critical cerebral vessel that supplies the pons, cerebellum, posterior brain regions, and inner ear. Potassium (K+) channel activity plays a significant role in determining vascular tone by regulating the cell membrane potential. Activation of inward rectifying K+ (Kir) channels, like other K+ channels, leads to cell membrane hyperpolarization and vasodilation. In this study, freshly isolated smooth muscle cells from the basilar artery were used to record Kir currents via the whole-cell patch clamp technique. The effects of 100 µmol/L BaCl2, a Kir channel inhibitor, and 10 µmol/L sodium nitroprusside (SNP), a nitro vasodilator, on Kir channel currents were investigated. The results demonstrated that BaCl2 inhibited Kir channel currents in basilar artery smooth muscle cells, whereas SNP enhanced these currents. This protocol provides a comprehensive guide for preparing freshly isolated arterial smooth muscle cells and recording Kir channel currents using the patch clamp technique, offering a valuable resource for researchers seeking to master this method.

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膜片钳技术记录新近分离的基底动脉平滑肌细胞内向内整流K+电流。
脑血管病是老年人的常见病,发病率稳步上升。基底动脉是一条重要的脑血管,供应脑桥、小脑、脑后区和内耳。钾离子通道活性通过调节细胞膜电位在血管张力中起重要作用。向内整流K+ (Kir)通道的激活,像其他K+通道一样,导致细胞膜超极化和血管舒张。在这项研究中,从基底动脉新鲜分离的平滑肌细胞通过全细胞膜片钳技术记录Kir电流。研究了100µmol/L的Kir通道抑制剂BaCl2和10µmol/L的硝基血管扩张剂硝普钠(SNP)对Kir通道电流的影响。结果表明,BaCl2抑制基底动脉平滑肌细胞的Kir通道电流,而SNP增强了这些电流。该方案为制备新鲜分离的动脉平滑肌细胞和使用膜片钳技术记录Kir通道电流提供了全面的指导,为寻求掌握该方法的研究人员提供了宝贵的资源。
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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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