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

Article DOI: 10.3791/58043-v • 09:33 min • June 26th, 2018
June 26th, 2018 •
1, 1, 1, 1
1Institute for Anatomy and Embryology, University Medical Centre Göttingen

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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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Optical Assay Synaptic Vesicle Recycling Cultured Neurons Overexpressing Presynaptic Proteins Antibodies Specificity Michela Matteoli Effectiveness Protein Cordocti Localized Synapses Primary Hippocampal Cells Coverslips 24-well Plate Reduced-serum Medium Cell-culture Medium Distilled Water Water Bath Laminar Flow Hood Sterile Working Conditions Transfection Mix Calcium Chloride Endotoxin-free DNA Transfection Buffer Best Results
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