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Kinetics of Synaptic Vesicle Refilling with Neurotransmitter Glutamate

机译：神经递质谷氨酸补充突触囊泡的动力学

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After releasing neurotransmitter, synaptic vesicles are retrieved by endocytosis and recycled via fast and slow pathways to be reused for synaptic transmission. To maintain the synaptic efficacy, vesicles must be refilled with neurotransmitter during recycling. However, the refilling speed estimated in isolated or reconstructed vesicles is, thus far, too slow to fill up vesicles within the period of recycling. Here, we re-examined the vesicle refilling rate directly at central glutamatergic synapses in slices, using simultaneous presynaptic and postsynaptic whole-cell recording combined with caged glutamate photolysis. After washing out vesicular glutamate, refilling of vesicles with uncaged glutamate caused a recovery of EPSCs with a time constant of 15 s that varied depending upon temperature, age, and cytosolic Cl - concentrations. This time constant is faster than that of the slow recycling pathway (～30 s) after clathrin-mediated endocytosis but is much too slow to fill up vesicles replenished through fast recycling pathways (1 s). After releasing neurotransmitter, synaptic vesicles are retrieved by endocytosis and refilled with neurotransmitter for reuse. Here, Hori and Takahashi determine the refilling time constant of glutamate into vesicles to be 15 s at room temperature at a CNS synapse.
机译：释放神经递质后，突触小泡通过内吞作用被回收，并通过快速和慢速途径再循环以重新用于突触传递。为了维持突触功效，在回收过程中必须用神经递质重新填充囊泡。然而，到目前为止，在分离的或重构的囊泡中估计的补充速度太慢，以致于在回收期间内无法充满囊泡。在这里，我们通过同时进行突触前和突触后全细胞记录结合笼状谷氨酸光解，直接检查了切片中中央谷氨酸能突触的囊泡充盈率。冲洗出囊状谷氨酸后，用未封入的谷氨酸重新填充囊泡，可恢复EPSC，其时间常数为15 s，该常数随温度，年龄和胞质Cl-浓度而变化。该时间常数比网格蛋白介导的内吞作用后的慢循环途径（〜30 s）要快，但太慢而无法填满通过快速循环途径（<1 s）补充的囊泡。释放神经递质后，通过内吞作用恢复突触小泡，并用神经递质重新填充以重新使用。在这里，Hori和Takahashi确定在室温下在CNS突触中谷氨酸向囊泡中补充的时间常数为15 s。

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