Single Ca2+ channels conducting 20 mM Ba2+ from adult rat dorsal root ganglion cells were characterized using the two-electrode patch-clamp technique configuration. 2. Channels demonstrating specific characteristics of conductance, voltage dependence and dihydropyridine sensitivity were classified as high-threshold or L-type Ca2+ channels. 3. Mean single-channel current in 20 mM Ba2+ did not show inactivation, but inactivation occurred when using Ca2+ as a permeating ion. 4. Stimulus protocols were delivered alternately in the cell-attached and whole-cell electrode, while recording single-channel activity and total Ca2+ current simultaneously. 5. A mean single-channel Ba2+ current from a stimulated patch did not show inactivation. However, stimulation of a physiological whole-cell Ca2+ current induced a marked inactivation of mean single-channel Ba2+ current. 6. Complete Ca2+ current block by the addition of 200 microM Cd2+ in the external solution removed single-channel inactivation in patches stimulated through a whole-cell electrode.
摘要
使用双电极膜片钳技术配置对成年大鼠背根神经节细胞中传导20 mM Ba2+的单个Ca2+通道进行了表征。2. 表现出特定电导、电压依赖性和二氢吡啶敏感性特征的通道被归类为高阈值或L型Ca2+通道。3. 20 mM Ba2+中的平均单通道电流未显示失活,但当使用Ca2+作为通透离子时会发生失活。4. 在细胞贴附式和全细胞膜片电极中交替施加刺激方案,同时记录单通道活性和总Ca2+电流。5. 来自受刺激膜片的平均单通道Ba2+电流未显示失活。然而,生理性全细胞Ca2+电流的刺激诱导了平均单通道Ba2+电流的明显失活。6. 通过在外部溶液中添加200 microM Cd2+完全阻断Ca2+电流,消除了通过全细胞膜片电极刺激的膜片中的单通道失活。