Dudel J, Franke C, Hatt H, Usherwood P N
Physiologisches Institut der Technischen Universität München, F.R.G.
Brain Res. 1989 Mar 6;481(2):215-20. doi: 10.1016/0006-8993(89)90796-8.
Outside-out patches of extrasynaptic membrane were isolated from leg muscles of locusts. L-Glutamate and its agonists were applied to such patches either continuously or in rapidly switched pulses. When the pipette contained a high chloride concentration, 2.5 x 10(-5) M glutamate triggered single-channel currents (gated by H-receptors) with a conductance of 25 pS which were carried by chloride, in addition to cationic channels (gated by D-receptors). For the chloride channels, the distribution of channel open times had components of about 2 and 12 ms. Pulses of higher glutamate concentrations elicited many superimposed channel openings, and the approximately saturating concentration of 10(-3) M glutamate opened 100-200 channels simultaneously. When the pipette contained low chloride, channel conductance was reduced, and the current voltage relation was shifted towards the now negative chloride equilibrium potential. H-Receptor-gated chloride channels were activated by glutamate, ibotenate and aspartate, but not by GABA, quisqualate, kainate, N-methyl-D-aspartate and carbachol. The currents declined in the continued presence of agonist showing a time constant of desensitization greater than 1 s. Recovery from desensitization after removal of the agonist was tested with double pulses and was found to have a time constant of about 300 ms.
从蝗虫腿部肌肉中分离出突触外膜的外翻膜片。将L-谷氨酸及其激动剂连续或快速切换脉冲施加于此类膜片上。当移液管中含有高浓度氯化物时,2.5×10⁻⁵ M的谷氨酸除了触发由D受体门控的阳离子通道外,还触发了由氯化物携带的、电导为25 pS的单通道电流(由H受体门控)。对于氯化物通道,通道开放时间的分布具有约2毫秒和12毫秒的成分。更高浓度谷氨酸的脉冲引发了许多叠加的通道开放,约10⁻³ M的谷氨酸饱和浓度同时打开了100 - 200个通道。当移液管中含有低浓度氯化物时,通道电导降低,电流 - 电压关系向现在为负的氯化物平衡电位移动。H受体门控的氯化物通道被谷氨酸、异博定和天冬氨酸激活,但不被GABA、quisqualate、 kainate、N - 甲基 - D - 天冬氨酸和卡巴胆碱激活。在激动剂持续存在的情况下,电流下降,脱敏时间常数大于1秒。用双脉冲测试了去除激动剂后脱敏的恢复情况,发现其时间常数约为300毫秒。