Network distribution of mitochondria and lipid droplets in human muscle fibres
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The objective of the present study was to develop a combination of fluorescent stains that would allow visualisation of the network of mitochondria and lipid droplets (intramyocellular lipids or IMCL) in human skeletal muscle fibres by means of conventional and confocal microscopy. Muscle biopsies were taken from the vastus lateralis of three lean, healthy and physically active male subjects. Frozen muscle sections were stained for mitochondria using antibodies against three mitochondrial proteins; porin, cytochrome c oxidase (COX) and NADH-ubiquinol oxidoreductase and neutral lipids were stained with oil red O. Anti-COX staining produced images with the strongest fluorescence signal and the highest resolution of the mitochondrial network and this stain was successfully combined with the antibody against type I fibre myosin. A highly organised matrix arrangement of mitochondria within the sarcomeres (in pairs at the I-band) was observed in the oxidative type I fibres. The density of mitochondria was the highest in the subsarcolemmal region. Anti-COX staining was combined with oil red O demonstrating that in type I fibres lipid droplets are mainly located in the space between the mitochondria.
KeywordsMitochondria Intramyocellular lipids Fluorescence Confocal imaging Skeletal muscle
The antibody against myosin (human slow fibres, A4.840) used in the study was developed by Dr. Blau and was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.
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