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用电压敏感染料测量新生大鼠心脏细胞培养束中的微观传导。

Microscopic conduction in cultured strands of neonatal rat heart cells measured with voltage-sensitive dyes.

作者信息

Fast V G, Kléber A G

机构信息

Department of Physiology, University of Bern Switzerland.

出版信息

Circ Res. 1993 Nov;73(5):914-25. doi: 10.1161/01.res.73.5.914.

DOI:10.1161/01.res.73.5.914
PMID:8403261
Abstract

Microscopic discontinuities in electrical activation were assessed in synthetic strands of neonatal rat myocytes cultured on a growth-directing matrix. An optical method using voltage-sensitive dye (RH-237) and a photodiode technique was used for recordings of membrane potential changes with subcellular resolution. Spatial resolution of the method (diameter of measurement area, 5.5 microns; interdiode distance, 30 microns) allowed for simultaneous measurements of cytoplasmic conduction time within a single cell and junctional conduction time across the cell border. In one-dimensional cell chains, where cells were juxtaposed by end-to-end connections but devoid of lateral connections, propagation of the excitation wave was strongly nonuniform: cytoplasmic conduction time was 38 +/- 30 (mean +/- SD) microseconds (n = 37), whereas junctional conduction time was 118 +/- 40 microseconds (n = 27, P < .0001). A mean delay introduced by a single junction was 80 microseconds, or 51% of conduction time. In two-dimensional strands consisting of several cells in width, which exhibited lateral as well as end-to-end connections, inhomogeneity of conduction was smaller: the cytoplasmic and junctional conduction times were 57 +/- 30 (n = 46) and 89 +/- 40 (n = 48) microseconds, respectively (P < .0001); mean junctional conduction delay was 32 microseconds (22% of conduction time). Mathematical modeling suggested that the averaging effect of lateral connections is caused by lateral convergence of local excitatory current beyond and lateral divergence before end-to-end connections. Our results demonstrate that the current flow through lateral cell-to-cell connections smooth the excitation wave front during longitudinal conduction in myocardial tissue.

摘要

在生长导向基质上培养的新生大鼠心肌细胞合成链中评估电激活的微观不连续性。使用电压敏感染料(RH - 237)的光学方法和光电二极管技术记录具有亚细胞分辨率的膜电位变化。该方法的空间分辨率(测量区域直径为5.5微米;二极管间距为30微米)允许同时测量单个细胞内的细胞质传导时间和跨细胞边界的连接传导时间。在一维细胞链中,细胞通过端对端连接并列但没有横向连接,兴奋波的传播非常不均匀:细胞质传导时间为38±30(平均值±标准差)微秒(n = 37),而连接传导时间为118±40微秒(n = 27,P <.0001)。单个连接引入的平均延迟为80微秒,占传导时间的51%。在由几个细胞宽度组成的二维链中,其表现出横向以及端对端连接,传导的不均匀性较小:细胞质和连接传导时间分别为57±30(n = 46)和89±40(n = 48)微秒(P <.0001);平均连接传导延迟为32微秒(传导时间的22%)。数学建模表明,横向连接的平均效应是由局部兴奋性电流在端对端连接之外的横向汇聚和端对端连接之前的横向发散引起的。我们的结果表明,在心肌组织纵向传导过程中,通过横向细胞间连接的电流流动使兴奋波前平滑。

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