Covarrubias M, Steinbach J H
Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110.
Pflugers Arch. 1990 Jun;416(4):385-92. doi: 10.1007/BF00370744.
Excision of membrane patches from clonal BC3H1 cells to the inside-out configuration reduces the mean open time and burst duration of skeletal muscle nicotinic acetylcholine receptors. The channel conductance is not altered. Our results indicate that the rates for several steps in receptor activation are affected. The altered receptors apparently form a homogeneous population. Patterns of activity similar to that seen after excision are seen infrequently in recordings from cell-attached patches, suggesting that receptors with this type of altered function are present in intact membranes. The extent and rate of the reduction in mean open time after excision are similar at acetylcholine concentration over a 1000-fold range, indicating that the alteration does not depend on the state of ligation of the receptor. The extent and rate also are similar at room temperature and at 11 degrees C. The changes seen indicate that careful comparisons to intact preparations are necessary when studying the gating of channels in excised patches or following reconstitution into artificial membranes.
从克隆的BC3H1细胞中切除膜片并转变为内面向外模式,会降低骨骼肌烟碱型乙酰胆碱受体的平均开放时间和爆发持续时间。通道电导未发生改变。我们的结果表明,受体激活过程中几个步骤的速率受到了影响。这些改变的受体显然构成了一个同质群体。在细胞贴附式膜片的记录中,很少见到与切除后相似的活动模式,这表明具有这种功能改变的受体存在于完整的膜中。在1000倍范围内的乙酰胆碱浓度下,切除后平均开放时间减少的程度和速率相似,这表明这种改变不依赖于受体的结合状态。在室温及11摄氏度时,其程度和速率也相似。所观察到的变化表明,在研究切除的膜片中通道的门控或重构到人工膜后的通道门控时,有必要与完整制剂进行仔细比较。