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人T细胞中钾离子通道特性的比较:穿孔膜片钳与全细胞记录技术

A comparison of K+ channel characteristics in human T cells: perforated-patch versus whole-cell recording techniques.

作者信息

Oleson D R, DeFelice L J, Donahoe R M

机构信息

Department of Psychiatry, Emory University, Atlanta, Georgia.

出版信息

J Membr Biol. 1993 Mar;132(3):229-41. doi: 10.1007/BF00235740.

Abstract

Standard whole-cell records using the patch-clamp technique are obtained after rupturing the cell membrane just below the patch pipette. Inherent problems, such as the disruption of cellular architecture and the displacement of cytosol, are unavoidable. In the present report, a whole-cell recording technique which makes use of a monovalent cation ionophore, nystatin, was applied to lymphocytes. Nystatin-perforated patches allow electrical access to the cell interior while virtually blocking the diffusion of cellular constituents into the electrode. By comparing standard whole-cell and perforated-patch techniques we observed marked differences in: activation, inactivation, and deactivation kinetics; steady-state inactivation; and the conductance-voltage relationship of K+ currents in activated human T cells.

摘要

使用膜片钳技术获得标准全细胞记录是在膜片吸管下方的细胞膜破裂后进行的。诸如细胞结构破坏和细胞质移位等内在问题是不可避免的。在本报告中,一种利用单价阳离子离子载体制霉菌素的全细胞记录技术被应用于淋巴细胞。制霉菌素穿孔膜片允许电信号进入细胞内部,同时几乎阻止细胞成分扩散到电极中。通过比较标准全细胞和穿孔膜片技术,我们观察到在激活、失活和去激活动力学;稳态失活;以及活化的人T细胞中K+电流的电导-电压关系方面存在显著差异。

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