Abstract
Mitochondrial function, a key indicator of cell health, can be assessed by monitoring changes in mitochondrial membrane potential (MMP). Cationic fluorescent dyes are commonly used tools to assess MMP. We used a water-soluble mitochondrial membrane potential indicator (m-MPI) to detect changes in MMP in HepG2 cells. A homogenous cell-based MMP assay was optimized and performed in a 1536-well plate format to screen several compound libraries for mitochondrial toxicity by evaluating the effects of chemical compounds on MMP.
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References
Chance B, Williams GR (1956) The respiratory chain and oxidative phosphorylation. Adv Enzymol Relat Subj Biochem 17:65–134
Mitchell P (1961) Coupling of phosphorylation to electron and hydrogen transfer by a chemi-osmotic type of mechanism. Nature 191:144–148
Chen LB (1988) Mitochondrial membrane potential in living cells. Annu Rev Cell Biol 4:155–181
Pieczenik SR, Neustadt J (2007) Mitochondrial dysfunction and molecular pathways of disease. Exp Mol Pathol 83(1):84–92
Lemasters JJ, Qian T, He L, Kim JS, Elmore SP, Cascio WE, Brenner DA (2002) Role of mitochondrial inner membrane permeabilization in necrotic cell death, apoptosis, and autophagy. Antioxid Redox Signal 4:769–781
Sakamuru S, Li X, Attene-Ramos MS, Huang R, Lu J, Shou L, Shen M, Tice RR, Austin CP, Xia M (2012) Application of a homogenous membrane potential assay to assess mitochondrial function. Physiol Genomics 44(9):495–503
Attene-Ramos MS, Huang R, Michael S, Witt KL, Richard A, Tice RR, Simeonov A, Austin CP, Xia M (2015) Profiling of the Tox21 chemical collection for mitochondrial function to identify compounds that acutely decrease mitochondrial membrane potential. Environ Health Perspect 123(1):49–56
Attene-Ramos MS, Huang R, Sakamuru S, Witt KL, Beeson GC, Shou L, Schnellmann RG, Beeson CC, Tice RR, Austin CP, Xia M (2013) Systematic study of mitochondrial toxicity of environmental chemicals using quantitative high throughput screening. Chem Res Toxicol 26(9):1323–1332
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Sakamuru, S., Attene-Ramos, M.S., Xia, M. (2016). Mitochondrial Membrane Potential Assay. In: Zhu, H., Xia, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 1473. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6346-1_2
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DOI: https://doi.org/10.1007/978-1-4939-6346-1_2
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6344-7
Online ISBN: 978-1-4939-6346-1
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