Nowycky M C, Fox A P, Tsien R W
Proc Natl Acad Sci U S A. 1985 Apr;82(7):2178-82. doi: 10.1073/pnas.82.7.2178.
A large-conductance calcium channel in chicken dorsal root ganglion neurons was studied with patch-clamp recordings of unitary currents. In addition to the conventional pattern of Ca-channel gating previously described in neurons ("mode 1"), we observed a different form of gating behavior ("mode 2"). Unlike the brief (approximately equal to 1 ms) openings in mode 1, mode 2 openings tend to be longer (greater than 10 ms) and often outlast the test pulse. In mode 2, the probability of channel openness (P) is high at relatively negative potentials where P in mode 1 is low. Mode 2 activity appears much less often than mode 1 activity in the absence of drug. However, the balance is strongly shifted in favor of mode 2 by the dihydropyridine Ca agonist Bay K 8644, an effect that underlies a marked enhancement of Ca-channel activity. This is the first evidence for dihydropyridine control of neuronal Ca-channel function at the single-channel level. Sweeps showing mode 1 or mode 2 gating appeared interspersed with sweeps with no openings, during which the channel was unavailable for opening ("null mode" or "mode 0"). Two approaches showed that switching between all three modes occurred on a time scale of seconds: (i) channels tended to remain in the same mode from one sweep to the next, with pulses at 0.25 Hz; and (ii) steady depolarizations in Bay K 8644 produced clusters of mode 2 openings lasting several seconds. Changes in the rates of switching might be important in neurochemical modulation of Ca channels. Bay K 8644 and other dihydropyridine Ca agonists might be useful experimental tools for manipulating transmitter release, neurite extension, and other neuronal functions dependent on intracellular Ca.
利用膜片钳记录单位电流的方法,对鸡背根神经节神经元中的一种大电导钙通道进行了研究。除了先前在神经元中描述的传统钙通道门控模式(“模式1”)外,我们还观察到了一种不同形式的门控行为(“模式2”)。与模式1中短暂的(约1毫秒)开放不同,模式2的开放往往持续时间更长(超过10毫秒),并且常常超过测试脉冲的持续时间。在模式2中,通道开放概率(P)在相对负电位时较高,而在模式1中该电位下P较低。在无药物情况下,模式2活动出现的频率远低于模式1活动。然而,二氢吡啶类钙激动剂Bay K 8644可使平衡明显偏向模式2,这一效应构成了钙通道活性显著增强的基础。这是二氢吡啶类药物在单通道水平上控制神经元钙通道功能的首个证据。显示模式1或模式2门控的扫描与无开放的扫描交替出现,在此期间通道无法开放(“零模式”或“模式0”)。两种方法表明,在数秒的时间尺度上会发生所有三种模式之间的转换:(i)通道在相邻扫描中往往保持在同一模式,脉冲频率为0.25赫兹;(ii)在Bay K 8644中进行持续去极化会产生持续数秒的模式2开放簇。转换速率的变化在钙通道的神经化学调制中可能很重要。Bay K 8644和其他二氢吡啶类钙激动剂可能是用于操纵递质释放、神经突延伸以及其他依赖细胞内钙的神经元功能的有用实验工具。