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THE LOCATION OF ADENINE NUCLEOTIDE IN THE STRIATED MUSCLE OF THE TOAD.

The location of tritiated adenine nucleotide was studied by autoradiography in toad muscle which had been fixed in an acetic acid/ethanol mixture, with lead acetate present to act as the nucleotide precipitant. The muscles were embedded in Araldite. The autoradiographs were examined in both the light microscope and the electron microscope. In confirmation of earlier work, it was found that at least 50 per cent of the adenine nucleotide is concentrated in the I band, near the A-I boundary, and evidence is now available to suggest that this fraction is located in the interfibrillar spaces rather than in the substance of the fibrils. Electron micrographs of unstained sections failed to show any structural features at the site in question, though the triads were made visible by the lead. However, when sections were stained with an alkaline solution containing lead, or with uranyl acetate, "vesicles" were revealed at the appropriate site and it is thought that these may be elements of a transverse network (reticulum) of tubules containing the adenine nucleotide. The location of phosphocreatine could not be investigated because this substance was lost from the muscle during the preparative procedure.

Adenine↗

EFFECTS OF EXTERNAL POTASSIUM AND STROPHANTHIDIN ON SODIUM FLUXES IN FROG STRIATED MUSCLE.

Unidirectional Na fluxes in isolated fibers from the frog's semitendinosus muscle were measured in the presence of strophanthidin and increased external potassium ion concentrations. Strophanthidin at a concentration of 10(-5)M inhibited about 80 per cent of the resting Na efflux without having any detectable effect on the resting Na influx. From this it is concluded that the major portion of the resting Na efflux is caused by active transport processes. External potassium concentrations from 2.5 to 7.5 mM had little effect on resting Na efflux. Above 7.5 mM and up to 15 mM external K, the Na efflux was markedly stimulated; with 15 mM K the Na influx was 250 to 300 per cent greater than normal. On the other hand, Na influx was unchanged with 15 mM K. The stimulated Na efflux with the higher concentrations was not appreciably reduced when choline or Li was substituted for external Na, but was completely inhibited by 10(-5)M strophanthidin. From these findings it is concluded that the active transport of Na is stimulated by the higher concentrations of K. It is postulated that this effect on the Na "pump" is produced as a result of the depolarization of the muscle membranes and is related to the increased metabolism and heat production found under conditions of high external K.

Animals↗

THE TIME COURSE OF THE LOSS AND RECOVERY OF CONTRACTURE ABILITY IN FROG STRIATED MUSCLE FOLLOWING EXPOSURE TO CA-FREE SOLUTIONS.

Using area under the contracture curve to quantitate contractures, the diffusion coefficient of calcium ions within the frog toe muscle during washout in a calcium-free solution and subsequent recovery after reintroduction of calcium to the bathing solution was calculated to be about 2 x 10(-6) cm(2)/sec. The diffusion coefficient measured during washout was found to be independent of temperature or initial calcium ion concentration. During recovery it was found to decrease if the temperature was lowered. This was likely due to the repolarization occurring after the depolarizing effect of the calcium-free solution. The relation between contracture area and [Ca](o) was found to be useful over a wider range than that between maximum tension and [Ca](o). The normalized contracture areas were larger at lower calcium concentrations if the contractures were produced with cold potassium solutions or if NO(3) replaced Cl in the bathing solutions. Decreasing the potassium concentration of the contracture solution to 50 mM from 115 mM did not change the relation between [Ca](o) and the normalized area. If the K concentration of the bathing solution was increased, the areas were decreased at lower concentrations of Ca.

Animals↗