Tour O, Parnas H, Parnas I
Otto Loewi Center for Cellular and Molecular Neurobiology, and Department of Neurobiology, The Hebrew University, Jerusalem, Israel.
Biophys J. 1998 Apr;74(4):1767-78. doi: 10.1016/S0006-3495(98)77887-7.
We have studied the voltage sensitivity of glutamate receptors in outside-out patches taken from crayfish muscles. We found that single-channel conductance, measured directly at the single-channel level, increases as depolarization rises. At holding potentials from -90 mV to approximately 20 mV, the conductance is 109 pS. At holding potentials positive to 20 mV, the conductance is 213 pS. This increase in single-channel conductance was also observed in cell-attached patches. In addition, desensitization, rise time, and the dose-response curve were all affected by depolarization. To further clarify these multifaceted effects, we evaluated the kinetic properties of single-channel activity recorded from cell-attached patches in hyperpolarization (membrane potential around -75 mV) and depolarization (membrane potential approximately 105 mV). We found that the glutamate dissociation rate constant (k_) was affected most significantly by membrane potential; it declined 6.5-fold under depolarization. The rate constant of channel closing (k(c)) was also significantly affected; it declined 1.8-fold. The rate constant of channel opening (k(o)) declined only 1.2-fold. The possible physiological significance of the depolarization-mediated changes in the above rate constants is discussed.
我们研究了从小龙虾肌肉获取的外翻式膜片中谷氨酸受体的电压敏感性。我们发现,在单通道水平直接测量的单通道电导会随着去极化程度的增加而增大。在 -90 mV 至约 20 mV 的钳制电位下,电导为 109 pS。在大于 20 mV 的钳制电位下,电导为 213 pS。在细胞贴附式膜片中也观察到了单通道电导的这种增加。此外,脱敏、上升时间和剂量反应曲线均受去极化影响。为了进一步阐明这些多方面的影响,我们评估了在超极化(膜电位约为 -75 mV)和去极化(膜电位约为 105 mV)条件下从细胞贴附式膜片记录的单通道活动的动力学特性。我们发现,谷氨酸解离速率常数(k_)受膜电位影响最为显著;在去极化条件下它下降了 6.5 倍。通道关闭速率常数(k(c))也受到显著影响;它下降了 1.8 倍。通道开放速率常数(k(o))仅下降了 1.2 倍。本文讨论了上述速率常数中去极化介导变化可能的生理意义。