JoVE Journal

Neuroscience

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Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain

5.9K Views

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12:14 min

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September 14th, 2022

DOI :

10.3791/64574-v

September 14th, 2022

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Sofia Massaro Tieze¹^,²^,³, Sreeganga S. Chandra¹^,², D. J. Vidyadhara¹^,²

¹Department of Neurology, Yale University, ²Department of Neuroscience, Yale University, ³Interdepartmental Neuroscience Program, Yale University

Summary

This protocol presents a robust, detailed method to obtain highly pure synaptosomes, synaptic vesicles, and other synaptic fractions from the mouse brain. This method enables the evaluation of synaptic processes, including the biochemical analysis of protein localization and function with compartmental resolution.

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Subcellular Fractionation

Synaptic Components

Synaptosome Structures

Molecular Processes

Brain Disorders

Protein Analysis

Subsynaptic Resolution

Tissue Collection

Experimental Procedure

Dissection Technique

Mid Sagittal Incision

External Acoustic Meatus

Chapters in this video

0:04

Introduction

1:37

Mouse Brain Excision

3:32

Synaptosome Preparation

5:23

Hypotonic Lysis

6:24

Synaptic Vesicle Isolation

7:35

Synaptic Plasma Membrane Isolation

10:02

Results: Electron Microscopy of Synaptosomes and Immunoblot Analysis of Subcellular Fractions

11:00

Conclusion

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