GSBS Student Publications

Title

Brain slice biotinylation: an ex vivo approach to measure region-specific plasma membrane protein trafficking in adult neurons

Student Author(s)

Luke R. Gabriel; Sijia Wu

GSBS Program

Neuroscience

UMMS Affiliation

Graduate School of Biomedical Sciences, Program in Neuroscience; Department of Psychiatry

Date

4-3-2014

Document Type

Article

Medical Subject Headings

Animals; Biotin; Brain; Brain Chemistry; Cell Membrane; Corpus Striatum; Dopamine Plasma Membrane Transport Proteins; Mice; Nerve Tissue Proteins; Neurons; Protein Kinase C; Protein Transport; Synaptic Transmission

Disciplines

Neuroscience and Neurobiology

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.

Rights and Permissions

Citation: J Vis Exp. 2014 Apr 3;(86). doi: 10.3791/51240. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Journal Title

Journal of visualized experiments : JoVE

PubMed ID

24747337