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

Bölümler

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

Özet

Otomatik Çeviri

Otomatik Çeviri

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.

Etiketler

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

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