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An Optical Assay for Synaptic Vesicle Recycling in Cultured Neurons Overexpressing Presynaptic Proteins
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Neurociência
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JoVE Journal Neurociência
An Optical Assay for Synaptic Vesicle Recycling in Cultured Neurons Overexpressing Presynaptic Proteins

An Optical Assay for Synaptic Vesicle Recycling in Cultured Neurons Overexpressing Presynaptic Proteins

DOI:
10.3791/58043-v

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09:33 min

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June 26, 2018

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, , ,
1Institute for Anatomy and Embryology,University Medical Centre Göttingen

Capítulos

  • 00:04Título
  • 00:31Transfection
  • 03:52Stimulation and Syt1 Uptake
  • 05:39Immunocytochemistry
  • 07:17Results: GFP-Rogdi at Functional Synapses
  • 08:42Conclusion

Summary

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We describe an optical assay for synaptic vesicle (SV) recycling in cultured neurons. Combining this protocol with double transfection to express a presynaptic marker and protein of interest allows us to locate presynaptic sites, their synaptic vesicle recycling capacity, and determine the role of the protein of interest.

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Keywords: Optical AssaySynaptic Vesicle RecyclingCultured NeuronsPresynaptic ProteinsAntibody SpecificityPrimary Hippocampal CellsTransfectionCalcium ChlorideEndotoxin-free DNATransfection BufferReduced-serum MediumCell-culture MediumIncubationWashingActive Synapses

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