title: Quantitative Microscopic Analysis of Plasma Membrane Receptor Dynamics in Living Plant Cells.
abstract: Plasma membrane-localized receptors are essential for cellular communication and signal transduction. In Arabidopsis thaliana, BRASSINOSTEROID INSENSITIVE1 (BRI1) is one of the receptors that is activated by binding to its ligand, the brassinosteroid (BR) hormone, at the cell surface to regulate diverse plant developmental processes. The availability of BRI1 in the plasma membrane is related to its signaling output and is known to be controlled by the dynamic endomembrane trafficking. Advances in fluorescence labeling and confocal microscopy techniques enabled us to gain a better understanding of plasma membrane receptor dynamics in living cells. Here we describe different quantitative microscopy methods to monitor the relative steady-state levels of the BRI1 protein in the plasma membrane of root epidermal cells and its relative exocytosis and recycling rates. The methods can be applied also to analyze similar dynamics of other plasma membrane-localized receptors.
authors.affiliation: Department of Plant Systems Biology, VIB, 9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Gent, Belgium.
authors.forename: Yu
authors.initials: Y
authors.lastname: Luo
authors.affiliation: VIB-Ghent University, Departments of Plant Systems Biology and Plant Biotechnology and Bioinformatics, 9052, Gent, Belgium. eurus@psb.vib-ugent.be.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Russinova
mesh_terms: D017360:Arabidopsis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D020126:Brefeldin A / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D059828:Plant Cells / Q000187:drug effects* / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D029681:Arabidopsis Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D020126:Brefeldin A; D003513:Cycloheximide; D011494:Protein Kinases; C108270:BRI1 protein, Arabidopsis
keywords: BRI1; Brassinosteroids; Confocal microscopy; Exocytosis; FRAP; Fluorescence; Plasma membrane; Recycling