The presence of a hyperpolarization-activated pacemaker (I(f)-like current was tested in dedifferentiated adult rat ventricular myocytes up to 12 days in primary culture with the whole-cell patch clamp technique. 2. An I(f)i-like current was found and characterized on freshly isolated and cultured ventricular cells. Both activation and density of the current varied in relation to the stage of dedifferentiation. The current was activated from -92.0 +/- 2.5 and -63 +/- 1.0 mV at the beginning (4-day-cultured cells) and end of the dedifferentiation process (12 days), respectively. Its density measured at 170 mV progressively increased from -2.34 +/- 0.36 to -6.12 +/- 0.64 pA pF-1 between the two farthest stages of cellular remodeling. In freshly isolated cells the current was activated at -108.0 +/- 1.5 mV and its current density measured at -170 mV was -1.97 +/- 0.56 pA pF-1. 3. The current was blocked by extracellular CsCl (3mM) in a voltage-dependent manner. Modification of reversal potentials obtained at various values of [K+]o ( 5.4 or 25 mM) and [Na+]o (140 or 30 mM) suggests that the current was carried by both K+ and Na+ ions. 4. It is concluded that the hyperpolarization-activated inward current, recorded in freshly isolated and in cultured ventricular cells has characteristics similar to those of I(f). In adult rat ventricular cells it is activated in a non-physiological potential range, but can be elicited in a more physiological range when the cells are remodelled through a dedifferentiated way. It is suggested that such a current could be implicated in ventricular arrhythmias developed in pathological events.
摘要
采用全细胞膜片钳技术,在原代培养长达12天的去分化成年大鼠心室肌细胞中检测超极化激活起搏器(类I(f)电流)的存在情况。2. 在新鲜分离和培养的心室细胞中发现并表征了类I(f)电流。该电流的激活和密度随去分化阶段而变化。在去分化过程开始时(培养4天的细胞)和结束时(12天),电流分别从-92.0±2.5 mV和-63±1.0 mV激活。在细胞重塑的两个最远阶段之间,在170 mV处测量的其密度从-2.34±0.36逐渐增加到-6.12±0.64 pA pF-1。在新鲜分离的细胞中,电流在-108.0±1.5 mV激活,在-170 mV处测量的电流密度为-1.97±0.56 pA pF-1。3. 该电流被细胞外CsCl(3 mM)以电压依赖性方式阻断。在不同的[K+]o(5.4或25 mM)和[Na+]o(140或30 mM)值下获得的反转电位的改变表明,该电流由K+和Na+离子共同携带。4. 得出结论,在新鲜分离和培养的心室细胞中记录到的超极化激活内向电流具有与I(f)相似的特征。在成年大鼠心室细胞中,它在非生理电位范围内激活,但当细胞通过去分化方式重塑时,可以在更生理的范围内引发。提示这种电流可能与病理事件中发生的室性心律失常有关。