微动脉段标本的全细胞膜片钳技术

山东大学学报(医学版) ›› 2011, Vol. 49 ›› Issue (3): 33-36.

微动脉段标本的全细胞膜片钳技术

李新芝1，司军强1,2，李丽1,2，赵磊1,2，马克涛1,2

石河子大学医学院 1.新疆地方与民族高发病教育部重点实验室电生理研究室；
2.生理学教研室，新疆石河子 832002

收稿日期:2010-07-07 出版日期:2011-03-10 发布日期:2011-03-10
通讯作者: 马克涛 (1979- ), 男, 副教授, 博士，主要从事血管电生理研究。 E-mail: maketao@hotmail.com
作者简介:李新芝(1980- ), 女, 讲师, 硕士，主要从事血管电生理研究。 E-mail: lixinzhi@shzu.edu.cn
基金资助:
国家自然科学基金资助项目(30960417, 81000411, 30900490 );兵团博士基金资助项目(2010GG34, 2010JC16);石河子大学高层次人才项目(RCZX200921);石河子大学自然科学与技术创新项目(ZRKX2009ZD03, ZRKK2009YB24)。

A method of whole-cell patch clamp recording from arteriolar segments

LI Xin-zhi1, SI Jun-qiang1,2, LI li1,2, ZHAO lei1,2, MA Ke-tao1,2

1. Department of Electrophysiology, Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Ministry of Education;
2. Department of Physiology, School of Medicine, Shihezi University, Shihezi 832002, Xinjiang, China

Received:2010-07-07 Online:2011-03-10 Published:2011-03-10

摘要/Abstract

摘要：

目的介绍一种在微动脉段标本上进行全细胞膜片钳记录的技术。方法在离体豚鼠耳蜗螺旋动脉(SMA)、小脑前下动脉(AICA)和肠系膜动脉(MA)分支微动脉(直径小于100μm)的原位平滑肌细胞上，应用全细胞膜片钳技术。结果微动脉段上平滑肌细胞静息膜电位平均值在-25 ~ -37mV之间，细胞膜电容(Cinput)平均值在70 ~250pF之间，均高于消化分离的单个平滑肌细胞。应用缝隙连接阻断剂DPBA(100μmol/L)后，SMA、AICA和MA微动脉段上平滑肌细胞膜电阻(Rinput)分别为（4937±741） MΩ(n=12)、（3703 ±367） MΩ(n=8)和（3336±479） MΩ(n=12)，细胞Cinput分别为（4.5±0.2） pF(n=9)、（7.1±0.7） pF(n=5)和（9.6±0.9） pF(n=7)，与单个平滑肌细胞相似，表明微动脉细胞间存在着丰富的缝隙连接。结论微动脉段标本全细胞膜片钳记录技术适用于微动脉细胞间缝隙连接、神经递质和药物对微动脉作用机制的研究。

关键词: 微动脉；缝隙连接；膜片钳；平滑肌细胞

Abstract:

Objective To introduce a method of whole-cell patch clamp recording from arteriolar segments. Methods Whole-cell patch clamp recordings were performed on smooth muscle cells (SMCs) in situ of isolated segments from guinea pig spiral modiolar arteries (SMA), anterior inferior cerebellar artery(AICA) branches and mesenteric artery(MA) branches (<100μm in diameter). Results The mean resting membrane potential of the three arterioles ranged from -25mV to -37mV, while the mean membrane capacitance (Cinput)of the three arterioles ranged from 70pF to 250pF, and both were much higher than those of the isolated single smooth muscle cell. After application of the gap junction blocker DPBA (100μmol/L), membrane resistance (Rinput) of cells in situ was (4937±741) MΩ(n=12), (3703±367) MΩ(n=8) and (3336±479) MΩ(n=12) in SMA, AICA and MA segments, respectively. The Cinput was (4.5±0.2) pF (n=9), (7.1±0.7) pF (n=5) and (9.6±0.9) pF (n=7), respectively. Both Rinput and Cinput were much closer to those of the isolated single smooth muscle cell, suggesting that gap junctions existed between cells of the arterioles. Conclusion Whole-cell patch clamp recording from cells in situ of arteriolar segments is potentially useful in the study of intercellular gap junctions and the mechanisms underlying the actions of neurotransmitters and drugs on arterioles.

Key words: Arteriole; Gap junction; Patch clamp; Smooth muscle cells

中图分类号:

R331.3

李新芝1，司军强1,2，李丽1,2，赵磊1,2，马克涛1,2. 微动脉段标本的全细胞膜片钳技术[J]. 山东大学学报(医学版), 2011, 49(3): 33-36.

LI Xin-zhi1, SI Jun-qiang1,2, LI li1,2, ZHAO lei1,2, MA Ke-tao1,2. A method of whole-cell patch clamp recording from arteriolar segments[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(3): 33-36.

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