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  • Protocol for Quantitative Analysis of Synaptic Vesicle Clustering in Axons of Cultured Neurons.

Protocol for Quantitative Analysis of Synaptic Vesicle Clustering in Axons of Cultured Neurons.

STAR protocols (2020-10-29)
Ji Won Um, Kyung Ah Han, Se-Young Choi, Jaewon Ko
摘要

Clustering of synaptic vesicles along the neuronal axons is a critical mechanism underpinning proper synaptic transmission. Here, we provide a detailed protocol for analyzing the distribution of synaptic vesicles in presynaptic boutons of cultured neurons. The protocol covers preparation of cultured neurons, expression of synaptic vesicle-enriched proteins, and quantification procedures. Utilizing neurons from postnatal transgenic mice, this method can be applied to investigate the roles of synaptic genes in regulating vesicle dynamics at synaptic sites. For complete details on the use and execution of this protocol, please refer to Han et al. (2020a).

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Triton X-100, for molecular biology
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蔗糖, ≥99.5% (GC), BioXtra
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氯化镁, anhydrous, ≥98%
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硼酸, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
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聚-D-赖氨酸 氢溴酸盐, mol wt 70,000-150,000
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牛血清白蛋白 来源于牛血清, heat shock fraction, pH 7, ≥98%