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Reactive oxygen species are important intracellular signaling molecules, but their mode of action is complex: In low concentrations they regulate key aspects of cellular function and behavior, while at high concentrations they can cause “oxidative stress”, which damages organelles, membranes and DNA. To analyze how redox signaling unfolds in single cells and organelles in real-time, an innovative optical microscopy technique has been developed jointly by the teams of LMU Professor Martin Kerschensteiner and TUM Professor Thomas Misgeld, both investigators of the Munich Cluster for Systems Neurology (SyNergy).
Kerschensteiner and Misgeld used redox-sensitive variants of the Green Fluorescent Protein (GFP) as visualization tools.
"By combining these with other biosensors and vital dyes, we were able to establish an approach that permits us to simultaneously monitor redox signals together with mitochondrial calcium currents, as well as changes in the electrical potential and the proton (pH) gradient across the mitochondrial membrane," says Thomas Misgeld.
Michael O. Breckwoldt, Franz Pfister, Peter M. Bradley, Petar Marinković, Philip R. Williams, Monika S. Brill, Barbara Plomer, Anja Schmalz, Daret K. St. Clair, Ronald Naumann, Oliver Griesbeck, Markus Schwarzländer, Leanne Godinho, Florence M. Bareyre, Tobias P. Dick, Martin Kerschensteiner and Thomas Misgeld, Multi-parametric optical analysis of mitochondrial redox signals during neuronal physiology and pathology, Nature Medicine (2014). DOI: 10.1038/nm.3520
Caption: The micrograph shows a peripheral nerve, with the neuromuscular endplates stained in red. The nerve-cell mitochondria were imaged with a fluorescent redox sensor (green in the cytoplasm, yellow at the endplates). Credit: M. Kerschensteiner and T. Misgeld