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A High-content Assay for Monitoring AMPA Receptor Trafficking

A High-content Assay for Monitoring AMPA Receptor Trafficking

Published on: January 28, 2019

Positioning of AMPA Receptor-Containing Endosomes Regulates Synapse Architecture

Marta Esteves da Silva¹, Max Adrian¹, Philipp Schätzle¹

¹Cell Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.

|November 14, 2015

Abstract:

Lateral diffusion in the membrane and endosomal trafficking both contribute to the addition and removal of AMPA receptors (AMPARs) at postsynaptic sites. However, the spatial coordination between these mechanisms has remained unclear, because little is known about the dynamics of AMPAR-containing endosomes. In addition, how the positioning of AMPAR-containing endosomes affects synapse organization and functioning has never been directly explored. Here, we used live-cell imaging in hippocampal neuron cultures to show that intracellular AMPARs are transported in Rab11-positive recycling endosomes, which frequently enter dendritic spines and depend on the microtubule and actin cytoskeleton. By using chemically induced dimerization systems to recruit kinesin (KIF1C) or myosin (MyosinV/VI) motors to Rab11-positive recycling endosomes, we controlled their trafficking and found that induced removal of recycling endosomes from spines decreases surface AMPAR expression and PSD-95 clusters at synapses. Our data suggest a mechanistic link between endosome positioning and postsynaptic structure and composition.

Keywords:

AMPA receptors KIF1C actin cytoskeleton dendritic spine intracellular transport kinesin microtubule myosin myosinV myosinVI synaptic plasticity

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