McCloskey Karen D
Centre for Biophotonics, University of Strathclyde, Glasgow, Scotland, UK.
BJU Int. 2006 Jun;97(6):1338-43. doi: 10.1111/j.1464-410X.2006.06156.x.
To determine whether there are inward currents in interstitial cells (IC) isolated from the guinea-pig detrusor and if so, to characterise them using the patch-clamp technique and pharmacological agents.
Using the whole-cell patch-clamp technique, inward currents were studied in IC enzymatically isolated from the detrusor of the guinea-pig bladder. Currents were evoked by stepping positively from a holding potential of - 80 mV.
Outward K+ currents were blocked by Cs+ internal solution to reveal inward currents, which activated at voltages more positive than - 50 mV, peaked at 0 mV, reversed near + 50 mV and were half-maximally activated at - 27 mV. The inward currents showed voltage-dependent inactivation and were half-maximally inactivated at - 36 mV. Fitting the activation and inactivation data with a Boltzmann function revealed a window current between - 40 mV and + 20 mV. The decay of the current evoked at 0 mV could be fitted with a single exponential with a mean time-constant of 88 ms. Replacing external Ca2+ with Ba2+ significantly increased this to 344 ms. The current amplitude was augmented by Ba2+, and by Bay K 8644. Inward currents were significantly reduced by 1 microm nifedipine, across the voltage range, but the blockade was more effective on the current evoked at 0 mV than that evoked by a step to - 20 mV, perhaps indicating voltage-dependence of the action of nifedipine or another component of inward current. Increasing the concentration of the drug to 10 microm caused no further significant reduction either at 0 mV or at -20 mV. However, in the presence of 1 microm nifedipine the latter current was significantly reduced by 100 microm Ni2+. Both currents were significantly reduced in Ca2+-free solution.
IC from the guinea-pig detrusor possess inward currents with typical characteristics of L-type Ca2+ current. They also have a component of inward Ca2+ current, which was resistant to nifedipine, but sensitive to Ni2+. Further work is needed to characterise the latter conductance.
确定从豚鼠逼尿肌分离出的间质细胞(IC)中是否存在内向电流,若存在,则使用膜片钳技术和药理学试剂对其进行特性描述。
采用全细胞膜片钳技术,研究从豚鼠膀胱逼尿肌酶解分离出的IC中的内向电流。电流通过从-80 mV的钳制电位正向阶跃来诱发。
外向K⁺电流被Cs⁺内液阻断以揭示内向电流,该内向电流在比-50 mV更正的电压下激活,在0 mV时达到峰值,在+50 mV附近反转,且在-27 mV时半最大激活。内向电流表现出电压依赖性失活,在-36 mV时半最大失活。用玻尔兹曼函数拟合激活和失活数据显示在-40 mV至+20 mV之间存在窗电流。在0 mV诱发的电流衰减可用单指数拟合,平均时间常数为88 ms。用Ba²⁺替代细胞外Ca²⁺可使其显著增加至344 ms。电流幅度被Ba²⁺和Bay K 8644增大。内向电流在整个电压范围内被1 μmol硝苯地平显著降低,但该阻断对在0 mV诱发的电流比对阶跃至-20 mV诱发的电流更有效,这可能表明硝苯地平作用或内向电流的另一成分具有电压依赖性。将药物浓度增加到10 μmol在0 mV或-20 mV时均未引起进一步的显著降低。然而,在存在1 μmol硝苯地平的情况下,后者的电流被100 μmol Ni²⁺显著降低。在无Ca²⁺溶液中两种电流均显著降低。
豚鼠逼尿肌的IC具有具有L型Ca²⁺电流典型特征的内向电流。它们还具有一部分内向Ca²⁺电流,该电流对硝苯地平耐药,但对Ni²⁺敏感。需要进一步的工作来描述后者的电导特性。