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J. Cell Biol.,
Volume 140, Number 5, March 9, 1998 1091-1099
* Neurobiochemisches Labor der Klinik für Neurologie; Confocal laser-scanning and digital fluorescence imaging microscopy were used to quantify the
mitochondrial autofluorescence changes of NAD(P)H
and flavoproteins in unfixed saponin-permeabilized myofibers from mice quadriceps muscle tissue. Addition of mitochondrial substrates, ADP, or cyanide led
to redox state changes of the mitochondrial NAD system. These changes were detected by ratio imaging of
the autofluorescence intensities of fluorescent flavoproteins and NAD(P)H, showing inverse fluorescence behavior. The flavoprotein signal was colocalized
with the potentiometric mitochondria-specific dye dimethylaminostyryl pyridyl methyl iodide (DASPMI),
or with MitoTrackerTM Green FM, a constitutive
marker for mitochondria. Within individual myofibers
we detected topological mitochondrial subsets with distinct flavoprotein autofluorescence levels, equally responding to induced rate changes of the oxidative phosphorylation. The flavoprotein autofluorescence levels
of these subsets differed by a factor of four. This heterogeneity was substantiated by flow-cytometric analysis
of flavoprotein and DASPMI fluorescence changes of
individual mitochondria isolated from mice skeletal
muscle. Our data provide direct evidence that mitochondria in single myofibers are distinct subsets at the
level of an intrinsic fluorescent marker of the mitochondrial NAD-redox system. Under the present experimental conditions these subsets show similar functional responses.
Arbeitsgruppe Molekulare Mustererkennung des
Instituts für Medizinische Neurobiologie; and § Institut für Medizinische Mikrobiologie, Universitätsklinikum der
Otto-von-Guericke-Universität, D-39120 Magdeburg, Germany
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