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Clcn3-/-鼠心肌细胞容积敏感性氯通道的电生理学研究(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2007年第1期
页码:
24-27
栏目:
基础研究
出版日期:
2007-01-01

文章信息/Info

Title:
Electrophysiology of volume sensitive chloride channel in Clcn3-/- mouse cardiomyocytes
作者:
龚卫琴1; 李源1; 王晓明1; 张珊红1; 臧益民2
第四军医大学：1.西京医院老年病科，2.基础部生理学教研室，陕西 西安 710032
Author(s):
GONG Weiqin1;  LI Yuan1;  WANG Xiaoming1;  ZHANG Shanhong1;  ZANG Yimin2
1.Department of Geriatrics, Xijing Hospital, 2.Department of Physiology, Fourth Military Medical University, Xi′an 710032, Shaanxi, China
关键词:
ClC3; 基因敲除; 心肌细胞; 容积敏感性氯电流
Keywords:
ClC3;  gene knockout;  cardiomyocytes;  volume sensitive chloride current
分类号:
R329.25
DOI:
-
文献标识码:
A
摘要:
目的 研究ClC3基因敲除（Clcn3-/-）鼠心肌细胞容积敏感性氯通道的电生理学特点。方法 制备Clcn3-/-鼠模型，采用膜片钳全细胞记录技术观察小鼠心肌细胞暴露于低渗溶液激活的容积敏感性氯电流及其蛋白激酶C（PKC）敏感性。结果 Clcn3-/-鼠心肌细胞在等渗状态下的背景电流很小，低渗刺激后的2.3±1.0 min膜电导改变不明显，随后增大的电流呈外向整流性、电压和时间依赖性部分失活，电流-电压曲线上的反转电位接近氯平衡电位的理论值，反转电位随细胞外氯浓度(［Cl-］o)降低向去极化电位方向偏移。氯通道阻断剂DIDS和NPPB可逆性抑制该通道的外向性膜电导，抑制强度分别为88.8%±2.6%和76.3%±2.1%。PKC激活剂PMA明显增大该电流，+100 mV和-100 mV增强幅度分别为110%±7%和110%±6%，而PKC抑制剂chelerythrine预处理能完全消除PMA的电流放大作用。结论 Clcn3-/-鼠心肌细胞存在典型的容积敏感性氯通道，PKC激活参与上调该通道的功能性表达。
Abstract:
AIM To study the electrophysiologic characteristics of volume sensitive chloride channel in Clcn3-/- mouse cardiomyocytes. METHODS Mice with homozygous disruption of Clcn3 gene (Clcn3-/-) were produced by techniques of gene targeting. The cardiomyocytes were isolated from adult Clcn3-/- mice and exposed to hypotonic solution. Whole cell patch clamp recording technique was used. RESULTS Small basal current was recorded under isotonic conditions. No significant alteration of the membrane conductance was observed during initial several minutes (2.3 ±1.0) min when the cells were exposed to hypotonic solution. The swelling-activated current exhibited moderate outward rectification and time-dependent inactivation at large positive potentials. The reversal potential of the current was close to the calculated equilibrium potential for ［Cl-］o. A rightward shift of the reversal potential was also observed upon reduction of ［Cl-］o. The chloride channel blockers DIDS and NPPB inhibited reversibly the outward conductance of the channel (by 88.8%±2.6% and 76.3%±2.1% suppression at +100 mV). PKC activator PMA significantly increased the swelling-activated current (by 110%±7% and 110%±6% amplitude respectively at +100 mV and -100 mV). Chelerythrine, a PKC inhibitor, eliminated completely the PMA effect on the chloride current. CONCLUSION The ClC3-deficient cardiomyocytes express the volume sensitive chloride channel. PKC activation upregulates the functional expression of the chloride channel.

参考文献/References

［1］Duan D, Winter C, Cowley S, et al. Molecular identification of a volume-regulated chloride channel［J］. Nature, 1997, 390(6658):417-421.

［2］Friedrich T, Breiderhoff T, Jentsch TJ. Mutational analysis demonstrates that ClC4 and ClC5 directly mediate plasma membrane currents［J］. J Biol Chem, 1999, 274(2):896-902.

［3］Weylandt KH, Valverde MA, Nobles M, et al. Human ClC3 is not the swellingactivated chloride channel involved in cell volume regulation［J］. J Biol Chem, 2001, 276(20):17461-17467.

［4］Stobrawa SM, Breiderhoff T, Takamori S, et al. Disruption of ClC3, a chloride channel expressed on synaptic vesicles, leads to a loss of the hippocampus［J］. Neuron, 2001, 29(1):185-196.

［5］Arreola J, Begenisich T, Nehrke K, et al. Secretion and cell volume regulation by salivary acinar cells from mice lacking expression of the Clcn3 Cl- channel gene［J］. J Physiol, 2002, 545(pt 1):207-216.

［6］Wang GX, Hatton WJ, Wang GL, et al. Functional effects of novel anti-ClC3 antibodies on native volumesensitive osmolyte and anion channels in cardiac and smooth muscle cells［J］. Am J Physiol Heart Circ Physiol, 2003, 285(4):H1453-H1463.

［7］Yoshikawa M, Uchida S, Ezaki J, et al. ClC3 deficiency leads to phenotypes similar to human neuronal ceroid lipofuscinosis［J］. Genes Cells, 2002, 7(6):597-605.

［8］Gong WQ, Xu HT,Shimizu T, et al. ClC3independent, PKC-dependent activity of volumesensitive Cl- channel in mouse ventricular cardiomyocytes［J］. Cell Physiol Biochem, 2004, 14(4-6):213-224.

［9］Okada Y, Oiki S, Hazama A, et al. Criteria for the molecular identification of the volumesensitive outwardly rectifying Cl- channel［J］. J Gen Physiol, 1998, 112(3):365-367.

［10］Hermoso M, Satterwhite CM, Andrade YN, et al. ClC3 is a fundamental molecular component of volumesensitive outwardly rectifying Cl- channels and volume regulation in HeLa cells and Xenopus laevis oocytes［J］. J Biol Chem, 2002, 277(42):40066-40074.

备注/Memo

备注/Memo:
收稿日期：2006-06-08.通讯作者: 臧益民, 教授, 主要从事心血管生理学的研究Email:zangym@fmmu.edu.cn 作者简介: 龚卫琴, 副主任医师Email:gongwq@fmmu.edu.cn
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