Cytochemical studies of adenosine triphosphatases in skeletal muscle fibers.
Mitochondrial ATPase and myosin ATPase have been localized in the muscle fibers of the rat diaphragm. The principal fiber type possesses a structure favorable for making this cytochemical separation with the light microscope. This small red fiber has numerous large, nearly spherical, mitochondria (ca. 1.5 micro) which are aggregated beneath the sarcolemma. In the interior of the fiber, smaller paired filamentous mitochondria (ca. 0.2 micro diameter) are aligned with the I band. Distribution of mitochondria was determined by sudanophilia, succinic dehydrogenase activity, and by direct examination with the electron microscope. ATPase activity at pH 7.2 is located in the large peripheral mitochondria and in the smaller mitochondria associated with the I band. The alignment of the small mitochondria results in a discrete cross-striated appearance in fibers stained for this enzymic activity. This mitochondrial ATPase does not cleave adenosine diphosphate or adenosine monophosphate; it is not sulfhydryl dependent and, in fact, is enhanced by the mercurial, p-hydroxymercuribenzoate. It requires magnesium ion and is stimulated by dinitrophenol. It is inhibited after formol-calcium fixation, but the residual activity is demonstrable by lengthening the incubation time. At pH 9.4 the ATPase is myofibrillar in origin and is located in the A bands. This myosin ATPase activity is sulfhydryl-dependent. Mercurial at this high pH has an interesting dual effect: it suppresses myosin ATPase but evokes mitochondrial ATPase activity. A third type of ATPase activity can be demonstrated, especially in the large white fibers. This activity occurs at pH 7.2 in the presence of cysteine. Its position is manifested cytochemically as a fine reticular pattern which surrounds individual myofibrils. The distribution suggests that it may originate in the sarcoplasmic reticulum.
D000251:Adenosine Triphosphatases; D000818:Animals; D002118:Calcium; D006651:Histocytochemistry; D006902:Hydroxymercuribenzoates; D008928:Mitochondria; D018485:Muscle Fibers, Skeletal; D009132:Muscles; D009210:Myofibrils; D009218:Myosins; D051381:Rats; D012519:Sarcoplasmic Reticulum
D006902:Hydroxymercuribenzoates; C030601:4-hydroxymercuribenzoate; D000251:Adenosine Triphosphatases; D009218:Myosins; D002118:Calcium
ADENOSINE TRIPHOSPHATASE; MUSCLES