Analysis of Rab protein function in neurotransmitter receptor trafficking at hippocampal synapses

Abstract

Members of the Rab family of small GTPases are essential regulators of intracellular membrane sorting. Nevertheless, very little is known about the role of these proteins in the membrane trafficking processes that operate at synapses, and specifically, at postsynaptic terminals. These events include the activity-dependent exocytic and endocytic trafficking of AMPA-type glutamate receptors, which underlies long-lasting forms of synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD). This chapter summarizes different experimental methods to address the role of Rab proteins in the trafficking of neurotransmitter receptors at postsynaptic terminals in the hippocampus. These techniques include immunogold electron microscopy to ultrastructurally localize endogenous Rab proteins at synapses, molecular biology methods to express recombinant Rab proteins in hippocampal slice cultures, electrophysiological techniques to evaluate the role of Rab proteins in synaptic transmission, and confocal fluorescence imaging to monitor receptor trafficking at dendrites and spines and its dependence on Rab proteins.