首页> 美国卫生研究院文献>The Journal of Physiology >Electrophysiology of single heart cells from the rabbit tricuspid valve.
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Electrophysiology of single heart cells from the rabbit tricuspid valve.

机译:来自兔三尖瓣的单个心脏细胞的电生理学。

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摘要

1. The electrophysiology of single myocytes isolated from the rabbit tricuspid valve was studied using the patch-clamp method (whole-cell configuration). Cell dispersion was achieved by collagenase treatment, using the Langendorff retrograde perfusion procedure. 2. After isolation, and while incubating in the recovery (Kraftbrühe) solution, cells had clear striations and were mostly spindle-shaped, or rod-like (less than 10%), with length varying from 35 microns to over 150 microns, and diameter from 3 to 10 microns. 3. Upon exposure to Tyrode solution, the calcium-tolerant cells were mostly rounded with smooth surfaces and well-defined borders. The mean diameter of these cells was 15 +/- 5 microns (S.D., n = 9). A smaller percentage (about 30%) retained the original elongated shape. 4. Patch pipette recordings showed the presence of spontaneous activity in about 30% of round cells, and less frequently in elongated cells. Maximum diastolic potentials (MDPs) in the round cells averaged -82 +/- 6 mV, with a take-off potential of -56 +/- 3 mV (n = 9), and an average maximum upstroke velocity (Vmax) value of 6.3 +/- 0.6 V/s (n = 4). In quiescent cells, the mean resting potential was 69 +/- 12 mV (n = 43). 5. Voltage clamp ramps revealed a steady-state I-V relation with a negative slope region. The mean input resistance value was 25 +/- 9 M omega (n = 16) for the elongated, and 883 +/- 481 M omega (n = 8) for the round cells. 6. Hyperpolarizing 5 s pulses (holding potential = -50 mV) occasionally revealed a slow, time-dependent inward current whose peak increased progressively as a function of clamp potential. The slowly activating current was sensitive to caesium 2 mM), indicating its similarity to the so-called 'pacemaker current' (iF). In alternate voltage- and current-clamp experiments, blocking of iF did not stop pacemaker activity, but there was up to a fourfold increase in pacemaker cycle length. 7. In some cells, 5 s hyperpolarizing steps from a holding potential of -40 or -50 mV produced large, inwardly directed and voltage-dependent current surges that decayed rapidly with time, similar to the inactivation described for the inward rectifier current, iK1. The current was very prominent at voltages more negative than -100 mV, and its decay process was best fitted by two time constants, one fast and one slow. For example, at -150 mV the time constants were 61 and 634 ms. The inward current was blocked by barium (1 mM).(ABSTRACT TRUNCATED AT 400 WORDS)
机译:1.使用膜片钳方法(全细胞配置)研究了从兔三尖瓣分离出的单个心肌细胞的电生理。使用Langendorff逆行灌注程序通过胶原酶处理实现细胞分散。 2.分离后,并在回收液(Kraftbrühe)中孵育时,细胞具有清晰的条纹,大部分呈纺锤形或棒状(小于10%),长度从35微米到150微米以上不等,并且直径从3到10微米。 3.暴露于Tyrode溶液中后,耐钙细胞大多呈圆形,表面光滑且边界清晰。这些细胞的平均直径为15 +/- 5微米(S.D.,n = 9)。较小的百分比(约30%)保留了原始的细长形状。 4.贴片移液器记录显示约30%的圆形细胞中存在自发活性,而细长细胞中较少。圆形细胞的最大舒张电位(MDP)平均为-82 +/- 6 mV,起飞电位为-56 +/- 3 mV(n = 9),平均最大上冲程速度(Vmax)值为6.3 +/- 0.6 V / s(n = 4)。在静态细胞中,平均静息电位为69 +/- 12 mV(n = 43)。 5.电压钳位斜坡显示出具有负斜率区域的稳态I-V关系。对于细长型电池,平均输入电阻值为25 +/- 9 MΩ(n = 16),对于圆形电池,平均输入电阻值为883 +/- 481 MΩ(n = 8)。 6.超极化5 s脉冲(保持电势= -50 mV)偶尔会显示出缓慢的,随时间变化的内向电流,其峰值随钳位电势而逐渐增加。缓慢激活的电流对2 mM的铯敏感,表明它与所谓的“起搏器电流”(iF)相似。在交替的电压和电流钳实验中,iF的阻滞并没有停止起搏器的活动,但是起搏器的周期长度增加了四倍。 7.在某些电池中,从-40或-50 mV的保持电势开始的5 s超极化步骤产生了大的,向内定向的和电压相关的电流浪涌,该浪涌随时间迅速衰减,类似于针对向内整流器电流iK1所述的失活。电流在负电压超过-100 mV时非常突出,并且其衰减过程最好由两个时间常数(一个快和一个慢)拟合。例如,在-150 mV时,时间常数分别为61和634 ms。内向电流被钡(1 mM)阻挡。(抽象截断为400字)

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