Ryanodine-induced stimulation of net Ca++ uptake by cardiac sarcoplasmic reticulum vesicles

Larry Jones, H. R. Besch, J. L. Sutko, J. T. Willerson

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Abstract

The effects of ryanodine on net Ca++ accumulation (Ca++ uptake) and on K+,Ca++-ATPase activities of cardiac sarcoplasmic reticulum vesicles were investigated. In the presence of 3 mM oxalate, ryanodine increased both the initial velocity of Ca++ uptake and the maximum amount of Ca++ accumulated 2- to 3-fold. Potassium ions potentiated the effects of ryanodine. In the presence of 100 mM KCl, 10-4 M ryanodine produced near maximal stimulation of uptake, whereas in the absence of KCl, a 10-fold higher concentration of ryanodine was required for appreciable stimulation. Choline chloride could not substitute for this effect of KCl. Ca++-dependent ATPase activities, whether measured with or without KCl, were not altered significantly by 10-4 M ryanodine when the concentration of Ca++ ions was not limiting and when oxalate was present. Thus, the stoichiometry of the net transport of Ca++ ions per molecule of ATP hydrolyzed was increased 3-fold by ryanodine to a value of approximately 1. Ryanodine stimulated net Ca++ accumulation at all Ca++ concentrations tested, but the effect was most marked at an ionized Ca++ concentration between 10-6 M and 10-5 M. The Mg++ concentration (1-6 mM), 3 mM ATP) had little effect on the stimulation of Ca++ uptake by 10-4 M ryanodine, and increasing the oxalate concentration to 6 mM did not alter the magnitude of stimulation of uptake by ryanodine. When oxalate was omitted from the incubation medium, K+,Ca++-ATPase activity was inhibited 50% by 10-4 M ryanodine, but this effect was completely reversed by addition of the Ca++-specific ionophore, A23187, which eliminated intravesicular accumulation of Ca++. The results suggest that ryanodine increases Ca++ uptake by cardiac sarcoplasmic reticulum vesicles by decreasing the rate of Ca++ efflux from the vesicles. No direct effect of the drug was observed on the turnover of the Ca++ pump or K+,Ca++-ATPase activity.

Original languageEnglish
Pages (from-to)48-55
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume209
Issue number1
StatePublished - 1979

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Ryanodine
Sarcoplasmic Reticulum
Oxalates
Adenosine Triphosphate
Ions
Ionophores
Ion Transport
Calcimycin
Choline
Adenosine Triphosphatases
Potassium

ASJC Scopus subject areas

  • Pharmacology

Cite this

Ryanodine-induced stimulation of net Ca++ uptake by cardiac sarcoplasmic reticulum vesicles. / Jones, Larry; Besch, H. R.; Sutko, J. L.; Willerson, J. T.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 209, No. 1, 1979, p. 48-55.

Research output: Contribution to journalArticle

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abstract = "The effects of ryanodine on net Ca++ accumulation (Ca++ uptake) and on K+,Ca++-ATPase activities of cardiac sarcoplasmic reticulum vesicles were investigated. In the presence of 3 mM oxalate, ryanodine increased both the initial velocity of Ca++ uptake and the maximum amount of Ca++ accumulated 2- to 3-fold. Potassium ions potentiated the effects of ryanodine. In the presence of 100 mM KCl, 10-4 M ryanodine produced near maximal stimulation of uptake, whereas in the absence of KCl, a 10-fold higher concentration of ryanodine was required for appreciable stimulation. Choline chloride could not substitute for this effect of KCl. Ca++-dependent ATPase activities, whether measured with or without KCl, were not altered significantly by 10-4 M ryanodine when the concentration of Ca++ ions was not limiting and when oxalate was present. Thus, the stoichiometry of the net transport of Ca++ ions per molecule of ATP hydrolyzed was increased 3-fold by ryanodine to a value of approximately 1. Ryanodine stimulated net Ca++ accumulation at all Ca++ concentrations tested, but the effect was most marked at an ionized Ca++ concentration between 10-6 M and 10-5 M. The Mg++ concentration (1-6 mM), 3 mM ATP) had little effect on the stimulation of Ca++ uptake by 10-4 M ryanodine, and increasing the oxalate concentration to 6 mM did not alter the magnitude of stimulation of uptake by ryanodine. When oxalate was omitted from the incubation medium, K+,Ca++-ATPase activity was inhibited 50{\%} by 10-4 M ryanodine, but this effect was completely reversed by addition of the Ca++-specific ionophore, A23187, which eliminated intravesicular accumulation of Ca++. The results suggest that ryanodine increases Ca++ uptake by cardiac sarcoplasmic reticulum vesicles by decreasing the rate of Ca++ efflux from the vesicles. No direct effect of the drug was observed on the turnover of the Ca++ pump or K+,Ca++-ATPase activity.",
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N2 - The effects of ryanodine on net Ca++ accumulation (Ca++ uptake) and on K+,Ca++-ATPase activities of cardiac sarcoplasmic reticulum vesicles were investigated. In the presence of 3 mM oxalate, ryanodine increased both the initial velocity of Ca++ uptake and the maximum amount of Ca++ accumulated 2- to 3-fold. Potassium ions potentiated the effects of ryanodine. In the presence of 100 mM KCl, 10-4 M ryanodine produced near maximal stimulation of uptake, whereas in the absence of KCl, a 10-fold higher concentration of ryanodine was required for appreciable stimulation. Choline chloride could not substitute for this effect of KCl. Ca++-dependent ATPase activities, whether measured with or without KCl, were not altered significantly by 10-4 M ryanodine when the concentration of Ca++ ions was not limiting and when oxalate was present. Thus, the stoichiometry of the net transport of Ca++ ions per molecule of ATP hydrolyzed was increased 3-fold by ryanodine to a value of approximately 1. Ryanodine stimulated net Ca++ accumulation at all Ca++ concentrations tested, but the effect was most marked at an ionized Ca++ concentration between 10-6 M and 10-5 M. The Mg++ concentration (1-6 mM), 3 mM ATP) had little effect on the stimulation of Ca++ uptake by 10-4 M ryanodine, and increasing the oxalate concentration to 6 mM did not alter the magnitude of stimulation of uptake by ryanodine. When oxalate was omitted from the incubation medium, K+,Ca++-ATPase activity was inhibited 50% by 10-4 M ryanodine, but this effect was completely reversed by addition of the Ca++-specific ionophore, A23187, which eliminated intravesicular accumulation of Ca++. The results suggest that ryanodine increases Ca++ uptake by cardiac sarcoplasmic reticulum vesicles by decreasing the rate of Ca++ efflux from the vesicles. No direct effect of the drug was observed on the turnover of the Ca++ pump or K+,Ca++-ATPase activity.

AB - The effects of ryanodine on net Ca++ accumulation (Ca++ uptake) and on K+,Ca++-ATPase activities of cardiac sarcoplasmic reticulum vesicles were investigated. In the presence of 3 mM oxalate, ryanodine increased both the initial velocity of Ca++ uptake and the maximum amount of Ca++ accumulated 2- to 3-fold. Potassium ions potentiated the effects of ryanodine. In the presence of 100 mM KCl, 10-4 M ryanodine produced near maximal stimulation of uptake, whereas in the absence of KCl, a 10-fold higher concentration of ryanodine was required for appreciable stimulation. Choline chloride could not substitute for this effect of KCl. Ca++-dependent ATPase activities, whether measured with or without KCl, were not altered significantly by 10-4 M ryanodine when the concentration of Ca++ ions was not limiting and when oxalate was present. Thus, the stoichiometry of the net transport of Ca++ ions per molecule of ATP hydrolyzed was increased 3-fold by ryanodine to a value of approximately 1. Ryanodine stimulated net Ca++ accumulation at all Ca++ concentrations tested, but the effect was most marked at an ionized Ca++ concentration between 10-6 M and 10-5 M. The Mg++ concentration (1-6 mM), 3 mM ATP) had little effect on the stimulation of Ca++ uptake by 10-4 M ryanodine, and increasing the oxalate concentration to 6 mM did not alter the magnitude of stimulation of uptake by ryanodine. When oxalate was omitted from the incubation medium, K+,Ca++-ATPase activity was inhibited 50% by 10-4 M ryanodine, but this effect was completely reversed by addition of the Ca++-specific ionophore, A23187, which eliminated intravesicular accumulation of Ca++. The results suggest that ryanodine increases Ca++ uptake by cardiac sarcoplasmic reticulum vesicles by decreasing the rate of Ca++ efflux from the vesicles. No direct effect of the drug was observed on the turnover of the Ca++ pump or K+,Ca++-ATPase activity.

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