Monitoring Activity-Dependent Bulk Endocytosis in Primary Neuronal Culture Using Large Fluorescent Dextrans
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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 wordsDextran Endocytosis Vesicle Neuron Presynapse
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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