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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/7815

Title: Measurement of cytosolic and mitochondrial pH in living cells during reversible metabolic inhibition
Authors: Balut, Corina
VAN DE VEN, Martin
Despa, S
Issue Date: 2008
Citation: KIDNEY INTERNATIONAL, 73(2). p. 226-232
Abstract: Renal ischemia and subsequent reperfusion lead to changes in the regulation of hydrogen ions across the mitochondrial membrane. This study was designed to monitor pH changes in the cytosol and mitochondria of Madin-Darby Canine Kidney cells exposed to metabolic inhibition and subsequent recovery. A classical one-photon confocal imaging approach using the pH-sensitive fluorophore carboxy SNARF-1 was used to define specific loading, calibration, and correction procedures to obtain reliable cytosolic and mitochondrial pH values in living cells. Metabolic inhibition resulted in both cytosolic and mitochondrial acidification, with a more pronounced decrease of mitochondrial pH as compared to the cytosolic pH. Shortly after removing the metabolic inhibition, cytosolic pH did not recover, whereas mitochondrial pH slowly increased. Our method is applicable to other cell types provided that the mitochondria can be loaded with SNARF-1 and that the cells possess a mitochondria-free region to measure SNARF-1 in the cytosol.
Notes: Hasselt Univ, Biomed Res Inst, Cell Physiol Grp, B-3590 Diepenbeek, Belgium. Transnatl Univ Limburg, Sch Life Sci, B-3590 Diepenbeek, Belgium. Int Ctr Biodynam, Dept Biophys, Bucharest, Romania. Loyola Univ Chicago, Dept Physiol, Maywood, IL USA. Hasselt Univ, Biomed Res Inst, Histol Grp, B-3590 Diepenbeek, Belgium.Smets, I, Hasselt Univ, Biomed Res Inst, Cell Physiol Grp, Agoralaan 1,Bldg D, B-3590 Diepenbeek, Belgium.ilse.smets@uhasselt.be
URI: http://hdl.handle.net/1942/7815
DOI: 10.1038/sj.ki.5002632
ISI #: 000252115200014
ISSN: 0085-2538
Category: A1
Type: Journal Contribution
Validation: ecoom, 2009
Appears in Collections: Research publications

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