Biomaterial Database

Simultaneous measurement of individual mitochondrial membrane potential and electrophoretic mobility by capillary electrophoresis. 2014

Gregory G Wolken, and Edgar A Arriaga

Department of Chemistry, University of Minnesota , 207 Pleasant Street SE, Minneapolis, Minnesota 55455, United States.

Mitochondrial membrane potential varies, depending on energy demand, subcellular location, and morphology and is commonly used as an indicator of mitochondrial functional status. Electrophoretic mobility is a heterogeneous surface property reflective of mitochondrial surface composition and morphology, which could be used as a basis for separation of mitochondrial subpopulations. Since these properties are heterogeneous, methods for their characterization in individual mitochondria are needed to better design and understand electrophoretic separations of subpopulations of mitochondria. Here we report on the first method for simultaneous determination of individual mitochondrial membrane potential and electrophoretic mobility by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). Mitochondria were isolated from cultured cells, mouse muscle, or liver, and then polarized, labeled with JC-1 (a ratiometric fluorescent probe, which indicates changes in membrane potential), and separated with CE-LIF. Red/green fluorescence intensity ratios from individual mitochondria were used as an indicator of mitochondrial membrane potential. Reproducible distributions of individual mitochondrial membrane potential and electrophoretic mobility were observed. Analysis of polarized and depolarized regions of interest defined using red/green ratios and runs of depolarized controls allowed for the determination of membrane potential and comparison of electrophoretic mobility distributions in preparations containing depolarized mitochondria. Through comparison of these regions of interest, we observed dependence of electrophoretic mobility on membrane potential, with polarized regions of interest displaying decreased electrophoretic mobility. This method could be applied to investigate mitochondrial heterogeneity in aging or disease models where membrane potential is an important factor.

UI	MeSH Term	Description	Entries
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D051379	Mice	The common name for the genus Mus.	Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D053078	Membrane Potential, Mitochondrial	The voltage difference, normally maintained at approximately -180mV, across the INNER MITOCHONDRIAL MEMBRANE, by a net movement of positive charge across the membrane. It is a major component of the PROTON MOTIVE FORCE in MITOCHONDRIA used to drive the synthesis of ATP.	Delta Psi M,DeltaPsi M,DeltapsiM,Mitochondrial Membrane Potential,Mitochondrial Transmembrane Potential,M, DeltaPsi,Membrane Potentials, Mitochondrial,Mitochondrial Membrane Potentials,Mitochondrial Transmembrane Potentials,Transmembrane Potential, Mitochondrial,Transmembrane Potentials, Mitochondrial
D019075	Electrophoresis, Capillary	A highly-sensitive (in the picomolar range, which is 10,000-fold more sensitive than conventional electrophoresis) and efficient technique that allows separation of PROTEINS; NUCLEIC ACIDS; and CARBOHYDRATES. (Segen, Dictionary of Modern Medicine, 1992)	Capillary Zone Electrophoresis,Capillary Electrophoreses,Capillary Electrophoresis,Capillary Zone Electrophoreses,Electrophoreses, Capillary,Electrophoreses, Capillary Zone,Electrophoresis, Capillary Zone,Zone Electrophoreses, Capillary,Zone Electrophoresis, Capillary
D024202	Electrophoretic Mobility Shift Assay	An electrophoretic technique for assaying the binding of one compound to another. Typically one compound is labeled to follow its mobility during electrophoresis. If the labeled compound is bound by the other compound, then the mobility of the labeled compound through the electrophoretic medium will be retarded.	Gelshift Analysis,Mobility Shift Assay,Band Shift Mobility Assay,Bandshift Mobility Assay,EMSA Electrophoretic Technique,Gel Retardation Assay,Gel Shift Analysis,Supershift Mobility Assay,Analyses, Gel Shift,Analysis, Gel Shift,Assay, Bandshift Mobility,Assay, Gel Retardation,Assay, Mobility Shift,Assay, Supershift Mobility,Assays, Bandshift Mobility,Assays, Gel Retardation,Assays, Mobility Shift,Assays, Supershift Mobility,Bandshift Mobility Assays,EMSA Electrophoretic Techniques,Electrophoretic Technique, EMSA,Electrophoretic Techniques, EMSA,Gel Retardation Assays,Gel Shift Analyses,Mobility Assay, Bandshift,Mobility Assay, Supershift,Mobility Assays, Bandshift,Mobility Assays, Supershift,Mobility Shift Assays,Supershift Mobility Assays,Technique, EMSA Electrophoretic,Techniques, EMSA Electrophoretic