Novel method for monitoring surface membrane trafficking on hippocampal acute slice preparation.
Thomas-Crusells J, Vieira A, Saarma M, Rivera C
J Neurosci Methods 2003 May 30;125(1-2):159-66.
Program for Molecular Neurobiology, Institute of Biotechnology, University of Helsinki, Viikki Biocenter, FIN-00014, Helsinki, Finland.
Protein trafficking has attracted considerable attention as a potential regulator of neuronal plasticity. Therefore, it is of interest to study the mechanism involved in protein trafficking in experimental paradigms commonly used in this context. Here, we present a method for cell surface protein biotinylation in the acute hippocampal slice, the most commonly used preparation for electrophysiological recordings. We validated this procedure with two previously characterized cell surface receptors, glutamate receptor subunit A (GluR A) and the transferrin receptor (TfR). We observed a glutamate-dependent increase in the degradation of surface GluR A, whereas the TfR did not show significant degradation in the time window used. In addition, the presented method offers the opportunity to study processes such as internalisation and recycling, and can also be applied to examine the effect of normal and pathological patterns of activity on membrane protein trafficking in commonly used preparations for electrophysiological recordings.
