Bean B P
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115.
J Neurosci. 1990 Jan;10(1):1-10. doi: 10.1523/JNEUROSCI.10-01-00001.1990.
The concentration dependence and kinetics of ionic currents activated by extracellular adenosine 5'-triphosphate (ATP) were studied in voltage-clamped dorsal root ganglion neurons from rats and bullfrogs. About 40% of neurons of both species responded to ATP with an increase in membrane conductance. The ATP-activated currents were similar in the 2 species, except that currents in rat neurons desensitized faster. In bullfrog neurons, the conductance was half-maximally activated by about 3 microM ATP; at low concentrations, the conductance increased 3- to 7-fold for a doubling in [ATP], suggesting that several ATP molecules must bind in order to activate the current. A steeper concentration-response relationship than expected from 1:1 binding was also seen in rat neurons. The current activated quickly upon application of ATP and decayed quickly when ATP was removed. Activation kinetics were faster at higher [ATP], with time constants decreasing from about 200 msec at 0.3 microM ATP to about 10 msec at 100 microM ATP. Deactivation kinetics (tau approximately 100-200 msec) were independent of the ATP concentration. The rapid activation and deactivation make it seem likely that the ATP-activated current is mediated by direct ligand binding rather than by a second-messenger system. The experimental observations can be mimicked by a simple model in which ATP must bind to 3 identical, noninteracting sites in order to activate a channel. The potency and kinetics of ATP action were voltage-dependent, with hyperpolarization slowing deactivation and increasing ATP's potency. Deactivation kinetics were also sensitive to the concentration of external Ca, becoming faster in higher Ca.
在对大鼠和牛蛙背根神经节神经元进行电压钳制的实验中,研究了细胞外5'-三磷酸腺苷(ATP)激活的离子电流的浓度依赖性和动力学。两种动物约40%的神经元对ATP产生反应,膜电导增加。两种动物中ATP激活的电流相似,只是大鼠神经元中的电流脱敏更快。在牛蛙神经元中,约3 microM的ATP可使电导达到最大激活的一半;在低浓度时,[ATP]加倍,电导增加3至7倍,这表明必须结合几个ATP分子才能激活电流。在大鼠神经元中也观察到比1:1结合预期更陡峭的浓度-反应关系。施加ATP后电流迅速激活,去除ATP后迅速衰减。在较高的[ATP]下激活动力学更快,时间常数从0.3 microM ATP时的约200毫秒降至100 microM ATP时的约10毫秒。失活动力学(τ约为100 - 200毫秒)与ATP浓度无关。快速激活和失活表明ATP激活的电流可能是由直接配体结合介导的,而不是通过第二信使系统。一个简单的模型可以模拟实验观察结果,即ATP必须结合到3个相同的、不相互作用的位点才能激活通道。ATP作用的效能和动力学是电压依赖性的,超极化会减缓失活并增加ATP的效能。失活动力学也对外界Ca的浓度敏感,在较高Ca浓度下会变得更快。