Quandt F N, MacVicar B A
Neuroscience. 1986 Sep;19(1):29-41. doi: 10.1016/0306-4522(86)90003-5.
The patch clamp technique was used to analyze single channel currents in intact and excised patches of glial cell membranes grown in primary cultures from newborn rat brain. Glial cells were morphologically identified by immunohistochemical staining for glial fibrillary acidic protein. Outward currents due to single channels were observed in recordings from both intact and excised patches obtained from the cell body region. The channel responsible for these currents was preferentially permeable to K+ because the reversal potential for this current was correlated with changes in the potassium equilibrium potential, when experimentally altered. The single channel conductance was 25 pS when measured between -20 and +20 mV in solutions with physiological K+ concentrations (10 degrees C). Channel gating was dependent on both the internal Ca2+ concentration and the membrane potential. Either depolarization of the membrane patch, or the addition of increasing Ca2+ concentrations to the internal surface, increased the probability of channel opening. Tetraethylammonium reversibly blocked the channel whereas 4-aminopyridine had no effect. The characteristics exhibited by this channel indicate that a Ca2+-activated K+ channel is present in the membrane of astrocytes grown in culture. These results, combined with previous evidence for a voltage dependent Ca2+ channel, suggest a dynamic role for glial cells in controlling excitability in the central nervous system. Influx of Ca2+ upon depolarization would increase the membrane permeability to K+ and could increase the "buffering" capacity of glial cells for extracellular K+.
采用膜片钳技术分析新生大鼠脑原代培养的完整和分离的神经胶质细胞膜片上的单通道电流。通过对胶质纤维酸性蛋白进行免疫组织化学染色从形态学上鉴定神经胶质细胞。在从细胞体区域获得的完整和分离的膜片记录中均观察到单通道引起的外向电流。负责这些电流的通道对K+具有优先通透性,因为当实验改变时,该电流的反转电位与钾平衡电位的变化相关。在生理K+浓度(10℃)的溶液中,在-20至+20mV之间测量时,单通道电导为25pS。通道门控取决于内部Ca2+浓度和膜电位。膜片的去极化或向内表面添加增加的Ca2+浓度均会增加通道开放的概率。四乙铵可逆地阻断该通道,而4-氨基吡啶则无作用。该通道表现出的特性表明培养的星形胶质细胞膜中存在Ca2+激活的K+通道。这些结果与先前关于电压依赖性Ca2+通道的证据相结合,表明神经胶质细胞在控制中枢神经系统兴奋性方面具有动态作用。去极化时Ca2+的内流会增加膜对K+的通透性,并可能增加神经胶质细胞对细胞外K+的“缓冲”能力。