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Measurement of transmembrane potential and current in cardiac muscle: a new voltage clamp method.

机译:心肌中跨膜电位和电流的测量:一种新的电压钳方法。

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摘要

1. A single sucrose gap voltage clamp technique was developed to correct for artifacts of 'leakage' corrent and extracellular resistance making possible improved measurement of membrane current and membrane potential in cardiac muscle. 2. A fourth compartment termed 'guard gap' was added to the sucrose gap. The guard gap is maintained at the same potential as the Reinger pool, so that no extracellular leakage current can flow into the Ringer pool. Comparison of experimental results with the predictions of an idealized cable model indicates that the guard gap is effective in trapping leakage current. 3. The slow charging of membrane capacitance due to extracellular series resistance was accelerated by applying a 'pre-pulse' of the command potential past the final voltage clamp value. 4. A second technique, termed 'chopped current pulse clamp', was used to compensate for the extracellular resistance throughout the voltage clamp step. The applied current was turned on and off at a frequency of 0-5-2 kHz. The membrane potential sampled during the zero current phase was fed back through the clamp loop. 5. With either of these compensation techniques, the voltage and current traces settle to effectively constant values within 2-4 msec after initiation of a hyperpolarizing voltage clamp step from rest. 6. The membrane conductance measured by the prepulse and chopped current-pulse technique are equal and confirm a higher conductance at rest than during the plateau of the action potential. 7. The 'instantaneous' current-voltage relation of the membrane is linear during the plateau of the frog ventricular action potential.
机译:1.开发了一种单一的蔗糖间隙电压钳技术,以纠正“泄漏”现象和细胞外电阻的假象,从而有可能改善心肌膜电流和膜电位的测量。 2.在蔗糖间隙中加入了称为“保护间隙”的第四个区室。保护间隙与Reinger池保持在相同的电位,因此没有细胞外漏电流可以流入Ringer池。将实验结果与理想电缆模型的预测结果进行比较表明,保护间隙有效地捕获了泄漏电流。 3.通过施加命令电位的“预脉冲”超过最终电压钳位值,可以加速由于细胞外串联电阻导致的膜电容缓慢充电。 4.第二种技术,称为“斩波电流脉冲钳位”,用于补偿整个电压钳位步骤中的细胞外电阻。施加的电流以0-5-2 kHz的频率打开和关闭。在零电流阶段采样的膜电位通过钳位环反馈。 5.使用这两种补偿技术中的任何一种,在从静止状态开始执行超极化电压钳位步骤后,电压和电流曲线便会在2-4毫秒内稳定为有效的恒定值。 6.通过预脉冲和斩波电流脉冲技术测得的膜电导是相等的,并确认静止时的电导高于动作电位的平稳期。 7.在青蛙心室动作电位的稳定期,膜的“瞬时”电流-电压关系是线性的。

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