title: Analysis of plant mitochondrial function using fluorescent protein sensors.
abstract: Mitochondrial physiology sets the basis for function of the organelle and vice versa. While a limited range of in vivo parameters, such as oxygen consumption, has been classically accessible for measurement, a growing collection of fluorescent protein sensors can now give insights into the physiology of plant mitochondria. Nevertheless, the meaningful application of these sensors in mitochondria is technically challenging and requires rigorous experimental standards. Here we exemplify the application of three genetically encoded sensors to monitor glutathione redox potential, pH, and calcium in the matrix of mitochondria in intact plants. We describe current methods for quantitative imaging and analysis in living root tips by confocal microscopy and discuss methodological limitations.
authors.affiliation: Plant Energy Biology Lab, INRES - Chemical Signalling, University of Bonn, Bonn, 53113, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Wagner
authors.forename: Thomas
authors.initials: T
authors.lastname: Nietzel
authors.forename: Isabel
authors.initials: I
authors.lastname: Aller
authors.forename: Alex
authors.initials: A
authors.lastname: Costa
authors.forename: Mark D
authors.initials: MD
authors.lastname: Fricker
authors.forename: Andreas J
authors.initials: AJ
authors.lastname: Meyer
authors.forename: Markus
authors.initials: M
authors.lastname: Schwarzländer
mesh_terms: D017360:Arabidopsis / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D061848:Optical Imaging / Q000379:methods*; D010084:Oxidation-Reduction; D018517:Plant Roots / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D005978:Glutathione; D002118:Calcium