Activity-dependent interactions of NSF and SNAP at living synapses.

Activity-dependent interactions of NSF and SNAP at living synapses.
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DOI:
10.1016/j.mcn.2011.02.002
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发表时间:
2011-05
影响因子:
3.5
通讯作者:
Ordway, Richard W.
Ordway, Richard W.
中科院分区:
医学3区
文献类型:
--
作者:
Yu, Wenhua;Kawasaki, Fumiko;Ordway, Richard W.

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作为神经递质释放装置的核心组分,SNARE、NSF和SNAP介导突触前质膜的特定区域(称为活性区(AZ))内的神经递质填充的突触囊泡的融合。本研究将果蝇的遗传学方法与生物化学和实时成像方法相结合,为NSF和SNAP在神经递质释放中的体内行为和相互作用提供了新的见解。这项工作采用了一个温度敏感(TS)麻痹NSF突变,昏迷,表明NSF功能的破坏导致活动依赖性的重新分配的NSF和SNAP的突触前质膜和蛋白质复合物的积累,含有SNARE,NSF和SNAP的周围活动区(PAZ)地区。昏迷状态下的荧光共振能量转移(FRET)和光漂白后的荧光恢复(FRAP)研究表明,NSF和SNAP在活体突触前末梢内表现出活性依赖性的相互结合以及独特的相互作用和迁移率。这些观察结果扩展了目前的模型描述的空间组织的NSF,SNAP和SNARE蛋白在突触囊泡贩运。
As core components of the neurotransmitter release apparatus, SNAREs, NSF and SNAPs mediate fusion of neurotransmitter-filled synaptic vesicles within specialized regions of the presynaptic plasma membrane known as active zones (AZs). The present study combines genetic approaches in Drosophila with biochemical and live-imaging methods to provide new insights into the in vivo behavior and interactions of NSF and SNAP in neurotransmitter release. This work employs a temperature-sensitive (TS) paralytic NSF mutant, comatose, to show that disruption of NSF function results in activity-dependent redistribution of NSF and SNAP to periactive zone (PAZ) regions of the presynaptic plasma membrane and accumulation of protein complexes containing SNAREs, NSF and SNAP. Fluoresence Resonance Energy Transfer (FRET) and Fluorescence Recovery After Photobleaching (FRAP) studies in comatose revealed that NSF and SNAP exhibit activity-dependent binding to each other within living presynaptic terminals as well as distinctive interactions and mobilities. These observations extend current models describing the spatial organization of NSF, SNAP and SNARE proteins in synaptic vesicle trafficking.
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