Summary
The purpose of this study was to examine the effect of three classes of Ca2+ antagonists, diltiazem, verapamil and nifedipine on Na+-Ca2+ exchange mechanism in the sarcolemmal vesicles isolated from canine heart. Na+-Ca2+ exchange and Ca2+ pump (ATP-dependent Ca2+ uptake) activities were assessed using the Millipore filtration technique. sarcolemmal vesicles used in this study are estimated to consist of several subpopulations wherein 23% are inside-out and 55% are right side-out sealed vesicles in orientation. The affect of each Ca2+ antagonist on the Na+-dependent Ca2+ uptake was studied in the total population of sarcolemmal vesicles, in which none of the agents depressed the initial rate of Ca2+ uptake until concentrations of 10 μM were incubated in the incubation medium. However, when sarcolemmal vesicles were preloaded with Ca2+ via ATP-dependent Ca2+ uptake, cellular Ca2+ influx was depressed only by verapamil (28%) at 1 μM in the efflux medium with 8 mM Na+. Furthermore, inhibition of Ca2+ efflux by verapamil was more pronounced in the presence of 16 mM Na+ in the efflux medium. The order of inhibition was; verapamil > diltiazem > nifedipine. These results indicate that same forms of Ca2+-antagonist drugs may affect the Na+-Ca2+ exchange mechanism in the cardiac sarcolemmal vesicles and therefore we suggest this site of action may contribute to their effects on the myocardium.
Similar content being viewed by others
References
Reuter H: Calcium channel modulation by neurotransmitters, enzymes and drugs. Nature 301:569–573, 1983
Glossman H, Ferry DR, Coll A, Rombush M: Molecular pharmacology of the calcium channel: Evidence for subtypes, multiple drug-receptor sites, channel subunits, and development of a radioiodinated 1,4-dihydropyridine calcium channel label, [125I]iodipine.J Cardiovasc Pharmacol 6(suppl 4): S608–5621, 1984
Schwartz A, Taira N: Calcium channel-blocking drugs: a novel intervention for the treatment of cardiac disease. Circ Res 52 (suppl II): I1-I2, 1983
Bolger G, Gengo P, Klockowski R, Luchowski E, Siegel H, Janis RA, Triggle AM: Characterization of binding of the Ca2+-channel antagonist, [3H]nitrendipine, to guinea pig ileal smooth muscle. J Pharmacol Exp Ther 225:291–309, 1983
Glossman H, Ferry DR, Goll A, Strissnig J and Schober M: Calcium channels:basic properties as revealed by radioligand binding tudies. J Cardiovasc Pharmacol 7 (suppl 6):S20-S30, 1985
Carcia ML, Frank King V, Siegel PKS, Reuban JP and Kazorowski GJ: Binding of Ca2+ entry blockers to cardiac sarcolemmal membrane vesicles. J Biol Chem 261:8146–8157, 1986
Bostrom SL, Ljung B, Mardh S, Forsen S and Thulin E: Interaction of the antihypertensive drug felodipine with calmodulin. Nature 292:777–778, 1981
Pang DC, Sperelakis N: Nifedipine, diltiazem, bepridil and verapamil uptake into cardiac and smooth muscles. Eur J Pharmacol 87:199–207, 1983
Cramb G, Dow JW: Uptake of bepridil into isolated ventricular myocytes; Retention by actin. Biochem Pharmacol 32:227–231, 1983
Norman JA, Ansell J, Phillips MA: Dihydropyridine Ca2+ entry blockers selectively inhibit peak I cAMP phosphodiesterase. Eur J Pharmacol 93:107–112, 1983
Mas-Oliver J, Nayler WG: The effect of verapamil on Ca2+-transporting and Ca2+-ATPase activity of isolated cardiac sarcolemmal preparations. Br J Pharmacol 70:617–624, 1980
Reuter H: Exchange of calcium ions in the mammalian myocardium. Mechanism and physiological significance. Circ Res 34:599–605, 1974
Caroni P, Carafoli E: The Ca2+-pumping ATPase of heart sarcolemma. J Biol Chem 256:3263–3270, 1987
Phillipson KD, Nishimoto AY: Stimulation of Na+-Ca2+ exchange in cardiac sarcolemmal vesicles by proteinase pretreatment. Am J Physiol 243:C191-C195, 1982
Phillipson KD, Nishimoto AY: Stimulation of Na+-Ca2+ exchange in cardiac sarcolemmal vesicles by phospholipase D. J Biol Chem 259:16–19, 1984
Phillipson KD: Interaction of charged amphiphils with Na+-Ca2+ exchange in cardiac sarcolemmal vesicles. J Biol Chem 259:13999–14002, 1984
Phillipson KD, Bersohn MM, Nishimoto AY: Effect of pH on Na2+-Ca2+ exchange in canine cardiac sarcolemmal vesicles. Circ Res 50:287–293, 1982
Reeves JP, Bailey CA, Hale CC: Redox modification of sodium-calcium exchange activity in cardiac sarcolemmal vesicles. J Biol Chem 261:4948–4955, 1986
Van Amsterdam FTM and Zaagsma J: Modulation of ATP dependent Ca2+ extrusion and Na+/Ca2+ exchange acrose rat cardiac sarcolemma by calcium antagonists. Eur J Pharmacol 123:441–449, 1986
Takeo S, Elimban V, Dhalla NS: Modification of cardiac sarcolemmal Na+-Ca2+ exchange by diltiazem and verapamil. Can J Cardiol 1:131–138, 1985
David-Dufilho M, Aude DeVynck M, Kazda S, Meyer M: Stimulation by nifedipine of calcium transport by cardiac sarcolemmal vesicles from spontaneously hypertensive rats. Eur J Pharmacol 97:121–127, 1984
Pitts BJR: Stoichiometry of sodium-calcium exchange in cardia sarcolemmal vesicles. J Biol Chem 254:6232–6235, 1979
Lowry OH, Resenbrough NJ, Farr AL, Randall RJ: Protein measurement with folin phenol reagent. J Biol Chem 193:265–275, 1951
Taussky HH, Shorr E: A microcolorimetric method for the determination of inorganic phosphorus. J Biol Chem 202:675–685, 1953
Lamers JM, Stinis JT, Kort WJ, Hullsman WG: Biochemical studies on the sarcolemmal function in the hypertrophied rabbit heart. J Mol Cell Cardiol 10:235–248, 1978
Masters BSS, William Jr CH, Kamin H: The preparation and properties of microsomal TPNH-cytochrome C reductase from pig liver. In: Estabrook RW, Pullmann ME (eds) Method in Enzymology. New York Academic Press 1967: 10 pp 565–573
Wharton DC, Tzagoloff A: Cytochrome oxidase from beef heart mitochrondria. In: Estabrook RE, Pullmann ME (eds) Method in Enzymology. New York Academic Press 1967: 10 pp 245–250
Makino N, Dhurubarajan R, Elimban V, Beamish RE, Dhalla NS: Alteration of sarcolemmal Na+-Ca2+ exchange in catecholamine-induced cardiomyopathy. Can J Cardiol 1:225–232, 1985
Makino N, Jasmin G, Beamish RE, Dhalla NS: Sarcolemmal Na+-Ca2+ exchange during the development of genetically determined cardiomyopathy. Biochem Biophys Res Commun 133:491–497, 1985
Panagia V, Makino N, Gangly P, Dhalla NS: Inhibition of Na+-Ca2+ exchange in heart sarcolemmal vesicles by phosphatidylethanolamine N-methylation. Eur J Biochem 166:597–603, 1987
Makino N, Dhalla KS, Elimban V, Dhalla NS: Sarcolemmal Ca2+ transport in streptozotocin-induced diabetic cardiomyopathy. Am J Physiol 253:E202-E207, 1987
Lucian S: Reconstitution of the sodium-calcium exchanger from cardiac sarcolemmal vesicles. Biochem Biophy Acta 772:127–134, 1984
Caroni P, Carafoli E: The regulation of the Na+-Ca2+ exchange of heart sarcolemma. Eur J Biochem 132:451–460, 1983
Bers DM, Allen LH, Kim Y: Calcium binding to cardiac sarcolemmal vesicles: potential role as a modifier of contraction. Am J Physiol 251:C861-C871, 1986
Phillipson KD: Symmetry properties of the Na+-Ca2+ exchange mechanisms in cardiac sarcolemmal vesicles. Biochem Biophy Acta 821:367–376, 1985
Lamers JM, Gysouw KJ, Verdouw PD: Slow calcium channel blockers and calmodulin. Biochem Pharmacol 34:3837–3843, 1985
Erdreich A, Spanier R, Rahamimoff H: The inhibition of Na+-dependent Ca2+ uptake by verapamil in synaptic plasma membrane vesicles. Eur J Pharmacol 90:193–202, 1983
Vaghy PL, Johnson JD, Matlib MA, Wang T, Schwartz A: Selective inhibition of Na+-induced Ca2+ release from heart mitochondria by diltiazem and certain other Ca2+-antagonist drugs. J Biol Chem 257:6000–6002, 1982
Younes A, Constantin M: Effects of bepridil on mitochondrial ATPase reactions. Biochem Pharmacol 29:1135–1139, 1980
Colvin RA, Pearson N, Messineo FC, Katz AM: Effect of Ca2+-channel blockers on Ca2+ transport and Ca2+ ATPase in skeletal and cardiac sarcoplasmic reticulum vesicles. J Cardiovasc Pharmacol 4:935–941, 1982
Itoh H, Ishikawa T, Hidaka H: Effects on calmodulin of bepridil, an antianginal agents. J Pharmacol Exp Ther 230:737–741, 1984
Agre PD, Virshup D, Bennet V: Bepridil and cetiedil, vasodilator which inhibit Ca2+-dependent calmodulin interactions with erythrocyte membranes. J Clin Invest 74:812–820, 1984
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hata, T., Makino, N., Nakanishi, H. et al. Modulation of Na+-Ca2+ exchange in cardiac sarcolemmal vesicles by Ca2+ antagonists. Mol Cell Biochem 84, 65–76 (1988). https://doi.org/10.1007/BF00235194
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00235194