Optical single-channel recording:: imaging Ca2+ flux through individual N-type voltage-gated channels expressed in Xenopus oocytes

被引:34
作者
Demuro, A [1 ]
Parker, I [1 ]
机构
[1] Univ Calif Irvine, Dept Neurobiol & Behav, Irvine, CA 92697 USA
关键词
Xenopus oocyte; single-channel recording; N-type Ca2+ channel; confocal microscopy; SKELETAL-MUSCLE FIBERS; INOSITOL TRISPHOSPHATE; CARDIAC MYOCYTES; HIGH-RESOLUTION; CALCIUM SPARKS; HEART-MUSCLE; PATCH-CLAMP; MEMBRANE; RELEASE; CELLS;
D O I
10.1016/S0143-4160(03)00154-4
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Functional studies of single membrane ion channels were made possible by the introduction of the patch-clamp technique, which allows single-channel currents to be measured with unprecedented resolution. Nevertheless, patch clamping has some limitations: including the need for physical access of the patch pipette, possible disruption of local cellular architecture, inability to monitor multiple channels, and lack of spatial information. Here, we demonstrate the use of confocal fluorescence microscopy as a non-invasive technique to optically monitor the gating of individual Ca2+ channels. Near-membrane fluorescence signals track the gating of N-type Ca2+ channels with a kinetic resolution of about 10 ms, provide a simultaneous and independent readout from several channels, and allow their locations to be mapped with sub-micrometer spatial resolution. Optical single-channel recording should be applicable to diverse voltage- and ligand-gated Ca2+-permeable channels, and has the potential for high-throughput functional analysis of single channels. (C) 2003 Elsevier Ltd. All rights reserved.
引用
收藏
页码:499 / 509
页数:11
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