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Monitoring Activity-Dependent Bulk Endocytosis in Primary Neuronal Culture Using Large Fluorescent Dextrans

  • Michael A. Cousin
  • Karen J. SmillieEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2233)

Abstract

The efficient recycling of synaptic vesicles (SVs) during neuronal activity is central for sustaining brain function. During intense neuronal activity, the dominant mechanism of SV retrieval is activity-dependent bulk endocytosis (ADBE). Here, we describe a method to monitor ADBE in isolation from other SV endocytosis modes, via the uptake of large fluorescent fluid-phase markers in primary neuronal culture. Furthermore, we outline how to monitor ADBE using this approach across a field of neurons or in individual neurons.

Key words

Dextran Endocytosis Vesicle Neuron Presynapse 

Notes

Acknowledgments

This work was supported by grants awarded by Cure Huntington’s Disease Initiative (A-11210 and A-14021; to KS and MC) and the Wellcome Trust (204954/Z/16/Z to MC).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Centre for Discovery Brain Sciences, Hugh Robson Building, George SquareUniversity of EdinburghEdinburghUK
  2. 2.Muir Maxwell Epilepsy Centre, Hugh Robson Building, George SquareUniversity of EdinburghEdinburghUK
  3. 3.Simons Initiative for the Developing Brain, Hugh Robson Building, George SquareUniversity of EdinburghEdinburghUK

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