Determination of myoglobin concentration and oxidative capacity in cryostat sections of human and rat skeletal muscle fibres and rat cardiomyocytes.
Myoglobin plays various roles in oxygen supply to muscle mitochondria. It is difficult, and in some cases impossible, to study the relationship between the myoglobin concentration and the oxidative capacity of individual muscle cells because myoglobin has to be fixed in situ whereas determination of oxidative capacity, for example, succinate dehydrogenase activity, requires unfixed cryostat sections. We have investigated whether a vapour-fixation technique allows the use of serial sections to study the relationship between myoglobin and succinate dehydrogenase activity. The technique is used to study a rat soleus muscle, two human skeletal muscle biopsies and biopsies of two patients with chronic heart failure, and in a control and hypertrophied rat heart. Staining intensities were quantified by microdensitometry. The absorbance values were calibrated using sections cut from gelatine blocks containing known amounts of myoglobin. The results show that it is possible to use serial sections for the determination of the myoglobin concentration and succinate dehydrogenase activity, and indicate that myoglobin can lead to a substantial reduction (18-60%) of the extracellular oxygen tension required to prevent an anoxic core in muscle cells.
van Beek-Harmsen
Brechje J
BJ
Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center, Van der Boechorststraat 7, 1081 BT, Amsterdam, The Netherlands.
Bekedam
Martijn A
MA
Feenstra
H Maria
HM
Visser
Frans C
FC
van der Laarse
Willem J
WJ
D000818:Animals; D006332:Cardiomegaly / Q000378:metabolism / Q000473:pathology; D015687:Cell Hypoxia; D016616:Cryoultramicrotomy; D006333:Heart Failure / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008722:Methods; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000378:metabolism*; D018482:Muscle, Skeletal; D032383:Myocytes, Cardiac / Q000737:chemistry / Q000378:metabolism*; D009211:Myoglobin / Q000032:analysis* / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D013385:Succinate Dehydrogenase / Q000378:metabolism
D009211:Myoglobin; D013385:Succinate Dehydrogenase