title: Measuring membrane protein dynamics in neurons using fluorescence recovery after photobleach.
abstract: The use of genetically encoded fluorescent tags such as green fluorescent protein (GFP) as reporters to monitor processes in living cells has transformed cell biology. One major application for these tools has been to analyze protein dynamics in neurons. In particular, fluorescence recovery after photobleach (FRAP) of surface expressed fluorophore-tagged proteins has been instrumental to addressing outstanding questions about how neurons orchestrate the synaptic delivery of proteins. Here, we provide an overview of the methodology, equipment, and analysis required to perform, analyze, and interpret these experiments.
authors.affiliation: MRC Centre for Synaptic Plasticity, School of Biochemistry, Medical Sciences Building, University of Bristol, Bristol, United Kingdom.
authors.forename: Inmaculada M
authors.initials: IM
authors.lastname: González-González
authors.forename: Frédéric
authors.initials: F
authors.lastname: Jaskolski
authors.forename: Yves
authors.initials: Y
authors.lastname: Goldberg
authors.forename: Michael C
authors.initials: MC
authors.lastname: Ashby
authors.forename: Jeremy M
authors.initials: JM
authors.lastname: Henley
mesh_terms: D000818:Animals; D058948:Cell Tracking / Q000379:methods*; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D049452:Green Fluorescent Proteins