Measuring mitochondrial transmembrane potential by TMRE staining

Crowley, Lisa C., Christensen, Melinda E. and Waterhouse, Nigel J. (2016) Measuring mitochondrial transmembrane potential by TMRE staining. Cold Spring Harbor Protocols, 2016 12: 1092-1096. doi:10.1101/pdb.prot087361

Author Crowley, Lisa C.
Christensen, Melinda E.
Waterhouse, Nigel J.
Title Measuring mitochondrial transmembrane potential by TMRE staining
Journal name Cold Spring Harbor Protocols   Check publisher's open access policy
ISSN 1559-6095
Publication date 2016-12-01
Year available 2016
Sub-type Article (original research)
DOI 10.1101/pdb.prot087361
Open Access Status Not yet assessed
Volume 2016
Issue 12
Start page 1092
End page 1096
Total pages 5
Place of publication Cold Spring Harbor, NY, United States
Publisher Cold Spring Harbor Laboratory Press
Language eng
Abstract Adenosine triphosphate (ATP) is the main source of energy for metabolism. Mitochondria provide the majority of this ATP by a process known as oxidative phosphorylation. This process involves active transfer of positively charged protons across the mitochondrial inner membrane resulting in a net internal negative charge, known as the mitochondrial transmembrane potential (ΔΨm). The proton gradient is then used by ATP synthase to produce ATP by fusing adenosine diphosphate and free phosphate. The net negative charge across a healthy mitochondrion is maintained at approximately −180 mV, which can be detected by staining cells with positively charged dyes such as tetramethylrhodamine ethyl ester (TMRE). TMRE emits a red fluorescence that can be detected by flow cytometry or fluorescence microscopy and the level of TMRE fluorescence in stained cells can be used to determine whether mitochondria in a cell have high or low ΔΨm. Cytochrome c is essential for producing ΔΨm because it promotes the pumping the protons into the mitochondrial intermembrane space as it shuttles electrons from Complex III to Complex IV along the electron transport chain. Cytochrome c is released from the mitochondrial intermembrane space into the cytosol during apoptosis. This impairs its ability to shuttle electrons between Complex III and Complex IV and results in rapid dissipation of ΔΨm. Loss of ΔΨm is therefore closely associated with cytochrome c release during apoptosis and is often used as a surrogate marker for cytochrome c release in cells.
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Document type: Journal Article
Sub-type: Article (original research)
Collections: Mater Research Institute-UQ (MRI-UQ)
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