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来自兔三尖瓣的单个心脏细胞的电生理学

Electrophysiology of single heart cells from the rabbit tricuspid valve.

作者信息

Anumonwo J M, Delmar M, Jalife J

机构信息

Department of Pharmacology, SUNY/Health Science Center, Syracuse 13210.

出版信息

J Physiol. 1990 Jun;425:145-67. doi: 10.1113/jphysiol.1990.sp018097.

Abstract
  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)
摘要
  1. 采用膜片钳技术(全细胞模式)研究了从兔三尖瓣分离的单个心肌细胞的电生理学特性。通过胶原酶处理并运用Langendorff逆行灌注法实现细胞分散。

  2. 分离后,细胞在恢复(Kraftbrühe)溶液中孵育时,具有清晰的横纹,大多呈纺锤形或棒状(少于10%),长度从35微米到超过150微米不等,直径为3至10微米。

  3. 暴露于Tyrode溶液后,耐钙细胞大多呈圆形,表面光滑,边界清晰。这些细胞的平均直径为15±5微米(标准差,n = 9)。较小比例(约30%)的细胞保留了原来的细长形状。

  4. 膜片钳记录显示,约30%的圆形细胞存在自发活动,细长形细胞中较少见。圆形细胞的最大舒张电位(MDPs)平均为-82±6 mV,起始电位为-56±3 mV(n = 9),平均最大除极速度(Vmax)值为6.3±0.6 V/s(n = 4)。在静息细胞中,平均静息电位为69±12 mV(n = 43)。

  5. 电压钳斜坡显示出具有负斜率区域的稳态I-V关系。细长形细胞的平均输入电阻值为25±9 MΩ(n = 16),圆形细胞为883±481 MΩ(n = 8)。

  6. 超极化5秒脉冲(钳制电位=-50 mV)偶尔会显示出一种缓慢的、时间依赖性内向电流,其峰值随钳制电位的增加而逐渐增大。这种缓慢激活的电流对2 mM铯敏感,表明其与所谓的“起搏电流”(iF)相似。在交替的电压钳和电流钳实验中,阻断iF并没有停止起搏活动,但起搏周期长度增加了四倍。

  7. 在一些细胞中,从-40或-50 mV的钳制电位进行5秒超极化步骤会产生大的、内向且电压依赖性的电流激增,随时间迅速衰减,类似于内向整流电流iK1的失活。该电流在电压比-100 mV更负时非常显著,其衰减过程最适合用两个时间常数来拟合,一个快一个慢。例如,在-150 mV时,时间常数分别为61和634毫秒。内向电流被1 mM钡阻断。(摘要截断于400字)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7804/1189842/41b3d0a87c0c/jphysiol00465-0159-a.jpg

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