Gleason E, Borges S, Wilson M
Division of Biological Sciences, University of California, Davis 95616.
J Neurosci. 1993 Jun;13(6):2359-70. doi: 10.1523/JNEUROSCI.13-06-02359.1993.
We have examined synaptic transmission between isolated pairs of chick GABAergic amacrine cells, maintained in sparse culture and identified by their binding of an amacrine cell-selective antibody. Using the perforated-patch method to whole-cell clamp both cells of a pair, postsynaptic currents were examined for step depolarizations of the "presynaptic" cell. Synaptic transmission, frequently reciprocal, was calcium dependent and reversibly blocked by bicuculline. Post-synaptic currents, excluding those due to ohmic electrical coupling, were elicited only for presynaptic voltage steps positive to about -40 mV and were always very noisy, suggesting that they were summed from relatively small numbers of quanta. Postsynaptic currents continued well after the termination of the 100 msec presynaptic voltage step when the step was to -10 mV, or positive to this value. This result is interpreted to imply that presynaptic calcium concentration remains elevated after the membrane is returned to its holding potential. When presynaptic voltages were kept low or else presynaptic voltage was uncontrolled, spontaneous quantal events mediated by GABAA receptors could often be seen. Quanta rose quickly (less than 4 msec) and decayed with a mean time constant of 19.3 msec. The amplitude distributions of quantal currents were positively skewed, sometimes showing rare quanta of exceptionally large amplitude. Peak conductance per quantum was about 300 pS, corresponding to the simultaneous opening of only 17 GABAA channels and corresponding to a net flux of only 32 x 10(3) Cl- ions per millivolt of driving force. Estimates of the maximum sustained release rate at individual release sites suggest an upper bound of between 19 and 42 quanta per second.
我们研究了分离培养的鸡γ-氨基丁酸能无长突细胞对之间的突触传递,这些细胞维持在稀疏培养状态,并通过它们与无长突细胞选择性抗体的结合来鉴定。使用穿孔膜片钳方法对一对细胞进行全细胞钳制,检测“突触前”细胞阶跃去极化时的突触后电流。突触传递通常是双向的,依赖于钙,可被荷包牡丹碱可逆性阻断。排除欧姆电耦合引起的电流,突触后电流仅在突触前电压阶跃正向至约 -40 mV时才会引发,并且总是非常嘈杂,这表明它们是由相对少量的量子总和而成。当突触前电压阶跃至 -10 mV或更高时,在100毫秒突触前电压阶跃终止后,突触后电流仍持续良好。该结果被解释为意味着在膜回到其钳制电位后,突触前钙浓度仍保持升高。当突触前电压保持较低或突触前电压不受控制时,经常可以看到由GABAA受体介导的自发量子事件。量子上升迅速(小于4毫秒),平均衰减时间常数为19.3毫秒。量子电流的幅度分布呈正偏态,有时会出现极少数幅度异常大的量子。每个量子的峰值电导约为300 pS,仅对应于17个GABAA通道的同时开放,每毫伏驱动力仅对应32×10³个Cl⁻离子的净通量。对单个释放位点最大持续释放速率的估计表明,上限为每秒19至42个量子。