title: Brain slice biotinylation: an ex vivo approach to measure region-specific plasma membrane protein trafficking in adult neurons.
abstract: Regulated endocytic trafficking is the central mechanism facilitating a variety of neuromodulatory events, by dynamically controlling receptor, ion channel, and transporter cell surface presentation on a minutes time scale. There is a broad diversity of mechanisms that control endocytic trafficking of individual proteins. Studies investigating the molecular underpinnings of trafficking have primarily relied upon surface biotinylation to quantitatively measure changes in membrane protein surface expression in response to exogenous stimuli and gene manipulation. However, this approach has been mainly limited to cultured cells, which may not faithfully reflect the physiologically relevant mechanisms at play in adult neurons. Moreover, cultured cell approaches may underestimate region-specific differences in trafficking mechanisms. Here, we describe an approach that extends cell surface biotinylation to the acute brain slice preparation. We demonstrate that this method provides a high-fidelity approach to measure rapid changes in membrane protein surface levels in adult neurons. This approach is likely to have broad utility in the field of neuronal endocytic trafficking.
authors.affiliation: Program in Neuroscience, Graduate School of Biomedical Sciences, University of Massachusetts Medical School.
authors.forename: Luke R
authors.initials: LR
authors.lastname: Gabriel
authors.affiliation: Program in Neuroscience, Graduate School of Biomedical Sciences, University of Massachusetts Medical School.
authors.forename: Sijia
authors.initials: S
authors.lastname: Wu
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School; Haley.Melikian@umassmed.edu.
authors.forename: Haley E
authors.initials: HE
authors.lastname: Melikian
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry*; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003342:Corpus Striatum / Q000737:chemistry / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins / Q000032:analysis / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D011493:Protein Kinase C / Q000032:analysis / Q000378:metabolism; D021381:Protein Transport; D009435:Synaptic Transmission / Q000502:physiology
chemical_list: D050483:Dopamine Plasma Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D001710:Biotin; D011493:Protein Kinase C
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