title: Using total internal reflection fluorescence microscopy to observe ion channel trafficking and assembly.
abstract: Ion channels are integral membrane proteins that allow the flow of ions across membranes down their electrochemical gradients and are a major determinant of cellular excitability. They play an important role in a variety of biological processes as diverse as insulin release from beta cells in the pancreas through to cardiac and smooth muscle contraction. We have used total internal reflection fluorescence (TIRF) microscopy to watch ion channels being transported in vesicles along microtubules within the cytoplasm of the cell. Furthermore, we can directly observe the fusion of these vesicles with the plasma membrane and the release and radial dispersion of single ion channels into the membrane. Finally, automated single-particle tracking of these objects allowed us to determine their diffusional behavior.
authors.affiliation: Barts and The London School of Medicine & Dentistry, Queen Mary, University of London, London, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Schwarzer
authors.forename: Gregory I
authors.initials: GI
authors.lastname: Mashanov
authors.forename: Justin E
authors.initials: JE
authors.lastname: Molloy
authors.forename: Andrew
authors.initials: A
authors.lastname: Tinker
mesh_terms: D005456:Fluorescent Dyes / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051657:KCNQ1 Potassium Channel / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D021381:Protein Transport; D014162:Transfection
chemical_list: D005456:Fluorescent Dyes; D051657:KCNQ1 Potassium Channel