A novel method for monitoring surface membrane trafficking on hippocampal acute slice preparation.
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.
Thomas-Crusells
Judith
J
Program for Molecular Neurobiology, Institute of Biotechnology, University of Helsinki, Viikki Biocenter, FIN-00014, Helsinki, Finland.
Vieira
Amandio
A
Saarma
Mart
M
Rivera
Claudio
C
D000818:Animals; D030161:Avian Proteins; D019921:Biotinylation / Q000379:methods*; D015153:Blotting, Western; D004594:Electrophysiology / Q000379:methods; D018698:Glutamic Acid / Q000494:pharmacology; D031320:Hippocrateaceae / Q000502:physiology*; D006652:Histological Techniques; D066298:In Vitro Techniques; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008991:Monitoring, Physiologic / Q000379:methods*; D021381:Protein Transport / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000187:drug effects / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism; D013997:Time Factors
D030161:Avian Proteins; D018698:Glutamic Acid; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D018091:Receptors, AMPA; D011990:Receptors, Transferrin; C088981:estrogen-inducible transferrin receptor-like membrane glycoprotein, chicken