Abstract
The anti-arrhythmic effects of long-chain polyunsaturated fatty acids (PUFAs) may be related to their ability to alter calcium handling in cardiac myocytes. We investigated the effect of eicosapentanoic acid (EPA) and docosahexaenoic acid (DHA) on calcium sparks in rat cardiac myocytes and the effects of these PUFAs and the monounsaturated oleic acid on cardiac calcium release channels (RyRs). Visualization of subcellular calcium concentrations in single rat ventricular myocytes showed that intensity of calcium sparks was reduced in the presence of EPA and DHA (15 µM). It was also found that calcium sparks decayed more quickly in the presence of EPA but not DHA. Sarcoplasmic vesicles containing RyRs were prepared from sheep hearts and RyR activity was determined by either [3H]ryanodine binding or by single-channel recording. Bilayers were formed from phosphatidylethanolamine and phosphatidylcholine dissolved in either n-decane or n-tetradecane. EPA inhibited [3H]ryanodine binding to RyRs in SR vesicles with K I = 40 µM. Poly- and mono-unsaturated free fatty acids inhibited RyR activity in lipid bilayers. EPA (cytosolic or luminal) inhibited RyRs with K I =32 µM and Hill coefficient, n 1 = 3.8. Inhibition was independent of the n-alkane solvent and whether RyRs were activated by ATP or Ca2+. DHA and oleic acid also inhibited RyRs, suggesting that free fatty acids generally inhibit RyRs at micromolar concentrations.
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References
G.E. Billman H. Hallaq A. Leaf (1994) ArticleTitlePrevention of ischemia-induced ventricular fibrillation by omega 3 fatty acids. Proc. Natl. Acad. Sci. USA 91 4427–4430 Occurrence Handle1:CAS:528:DyaK2cXktVOqu74%3D Occurrence Handle8183925
G.E. Billman J.X. Kang A. Leaf (1997) ArticleTitlePrevention of ischemia-induced cardiac sudden death by n-3 polyunsaturated fatty acids in dogs. Lipids 32 1161–1168 Occurrence Handle1:CAS:528:DyaK2sXnsFaisb4%3D Occurrence Handle9397401
K.Y. Bogdanov H.A. Spurgeon T.M. Vinogradova E.G. Lakatta (1998) ArticleTitleModulation of the transient outward current in adult rat ventricular myocytes by polyunsaturated fatty acids. Am. J. Physiol. 274 H571–H579 Occurrence Handle1:CAS:528:DyaK1cXht1Cmtrg%3D Occurrence Handle9486261
M.L. Burr A.M. Fehily J.F. Gilbert S. Rogers R.M. Holliday P.M. Sweetnam P.C. Elwood N.M. Deadman (1989) ArticleTitleEffects of changes in fat, fish, and fibre intakes on death and myocardial reinfarction: diet and reinfarction trial. Lancet 8666 757–761 Occurrence Handle10.1016/S0140-6736(89)90828-3
M.C. Capogrossi S.R. Houser A. Bahinski E.G. Lakatta (1987) ArticleTitleSynchronous occurrence of spontaneous localized calcium release from the sarcoplasmic reticulum generates action potentials in rat cardiac ventricular myocytes at normal resting membrane potential. Circ. Res. 61 498–503 Occurrence Handle1:STN:280:BieD3c3mvVw%3D Occurrence Handle3652397
E.A. Capper L.A. Marshall (2001) ArticleTitleMammalian phospholipases A2: Mediators of inflammation, proliferation and apoptosis. Prog. Lipid Res. 40 167–197 Occurrence Handle10.1016/S0163-7827(01)00002-9 Occurrence Handle1:CAS:528:DC%2BD3MXislCktbg%3D Occurrence Handle11275266
W.E. Connor D.S. Lin C. Colvis (1996) ArticleTitleDifferential mobilization of fatty acids from adipose tissue. J. Lipid Res. 37 290–298 Occurrence Handle1:CAS:528:DyaK28XhsFClsrs%3D Occurrence Handle9026527
H.G.L. Coster D.R. Laver (1986) ArticleTitleThe effect of temperature on lipid/n-alkane interactions in lecithin bilayers. Biochim. Biophys. Acta 857 95–104 Occurrence Handle10.1016/0005-2736(86)90102-1 Occurrence Handle1:CAS:528:DyaL28Xit1Wjur4%3D Occurrence Handle3964707
M. Croset J.M. Black J.E. Swanson J.E. Kinsella (1989) ArticleTitleEffects of dietary n-3 polyunsaturated fatty acids on phospholipid composition and calcium transport in mouse cardiac sarcoplasmic reticulum. Lipids 24 278–285 Occurrence Handle1:CAS:528:DyaL1MXitlSmt74%3D Occurrence Handle2526915
M.C.G. Daniels D. Fedida C. Lamont H.E. ter Keurs (1991) ArticleTitleRole of the sarcolemma in triggered propagated contractions in rat cardiac trabeculae. Circ. Res. 68 1408–1421 Occurrence Handle1:STN:280:By6B3cfntlc%3D Occurrence Handle2018999
D.A. Ford (2002) ArticleTitleAlterations in myocardial lipid metabolism during myocardial ischemia and reperfusion. Prog. Lipid Res. 41 6–26 Occurrence Handle10.1016/S0163-7827(01)00014-5 Occurrence Handle1:CAS:528:DC%2BD3MXptlGjt7k%3D Occurrence Handle11694267
S. Grimsgaard K.H. Bonaa J-B. Hasen A. Nordoy (1997) ArticleTitleHighly purified eicosapentaenoic acid and docosahexaenoic acid in humans have similar triacylglycerol-lowering effects but divergent effects on serum fatty acids. Am. J. Clin. Nutr. 66 649–659 Occurrence Handle1:CAS:528:DyaK2sXlslKhs7s%3D Occurrence Handle9280188
B. Hille (1977) ArticleTitleLocal anesthetics: hydrophilic and hydrophobic pathways for the drug–receptor reaction. J. Gen. Physiol. 69 497–515 Occurrence Handle1:CAS:528:DyaE2sXhvVejsLc%3D Occurrence Handle300786
C.E. Hock L.D. Beck R.C. Bodine D.K. Reibel (1990) ArticleTitleInfluence of dietary n-3 fatty acids on myocardial ischemia and reperfusion. Am. J. Physiol. 259 H1518–H1526 Occurrence Handle1:CAS:528:DyaK3MXit1yhsg%3D%3D Occurrence Handle2240249
B.N. Honen D.A. Saint (2002) ArticleTitlePolyunsaturated dietary fats change the properties of calcium sparks in adult rat atrial myocytes. J. Nutr. Biochem. 13 322–329 Occurrence Handle10.1016/S0955-2863(02)00175-4 Occurrence Handle1:CAS:528:DC%2BD38XkslKitLk%3D Occurrence Handle12088797
E. Honore J. Barhanin B. Attali F. Lesage M. Lazdunski (1994) ArticleTitleExternal blockade of the major cardiac delayed-rectifier K+ channel (Kv1.5) by polyunsaturated fatty acids. Proc. Natl. Acad. Sci. USA 91 1937–1941 Occurrence Handle1:CAS:528:DyaK2cXitVyhtbw%3D Occurrence Handle8127910
D. Jiang B. Xiao L. Zhang S.R. Chen (2002) ArticleTitleEnhanced basal activity of a cardiac Ca2+ release channel (ryanodine receptor) mutant associated with ventricular tachycardia and sudden death. Circ. Res. 91 218–225 Occurrence Handle10.1161/01.RES.0000028455.36940.5E Occurrence Handle1:CAS:528:DC%2BD38Xmt12jtb4%3D Occurrence Handle12169647
J.X. Kang A. Leaf (1996) ArticleTitleProtective effects of free polyunsaturated fatty acids on arrhythmias induced by lysophosphatidylcholine or palmitoylcarnitine in neonatal rat cardiac myocytes. Eur. J. Pharmacol. 297 97–106 Occurrence Handle10.1016/0014-2999(95)00701-6 Occurrence Handle1:CAS:528:DyaK28Xht1WltL4%3D Occurrence Handle8851173
R.S. Kass W.J. Lederer R.W. Tsien R. Weingart (1978) ArticleTitleRole of calcium ions in transient inward currents and after contractions induced by strophanthidin in cardiac Purkinje fibres. J. Physiol. 281 187–208 Occurrence Handle1:CAS:528:DyaE1cXlvFSrsbk%3D Occurrence Handle702368
D. Kromhout E.B. Bosschieter C. de Lezenne Coulander (1985) ArticleTitleThe inverse relation between fish consumption and 20-year mortality from coronary heart disease. N. Engl. J. Med. 312 1205–1209 Occurrence Handle1:STN:280:BiqC1cfgvVQ%3D Occurrence Handle3990713
H. Kusuoka J.K. Porterfield H.F. Weisman M.L. Weisfeldt E. Marban (1987) ArticleTitlePathophysiology and pathogenesis of stunned myocardium: depressed Ca2+-activation as a consequence of reperfusion-induced cellular calcium overload in ferret hearts. J. Clin. Invest. 79 950–961 Occurrence Handle1:CAS:528:DyaL2sXitFWrtLw%3D Occurrence Handle3818956
D.R. Laver L.D. Roden G.P. Ahern K.R. Eager P.R. Junankar A.F. Dulhunty (1995) ArticleTitleCytoplasmic Ca2+ inhibits the ryanodine receptor from cardiac muscle. J. Membrane Biol. 147 7–22 Occurrence Handle1:CAS:528:DyaK2MXotVOiu7Y%3D
D.R. Laver K.R. Eager L. Taoube G.D. Lamb (2000) ArticleTitleEffects of cytoplasmic and luminal pH on Ca2+ release channels from rabbit skeletal muscle. Biophys. J 78 1835–1851 Occurrence Handle1:CAS:528:DC%2BD3cXisVWgt70%3D Occurrence Handle10733964
A. Leaf J.X. Kang Y.F. Xiao G.E. Billman R.A. Voskuyl (1999) ArticleTitleExperimental studies on antiarrhythmic and antiseizure effects of polyunsaturated fatty acids in excitable tissues. J. Nutr. Biochem. 10 440–448 Occurrence Handle10.1016/S0955-2863(99)00034-0 Occurrence Handle1:CAS:528:DyaK1MXls12ksrs%3D
W.R. Leifert E.J. McMurchie D.A. Saint (1999) ArticleTitleInhibition of cardiac sodium currents in adult rat myocytes by n-3 polyunsaturated fatty acids. J. Physiol. 520 671–679 Occurrence Handle1:CAS:528:DyaK1MXns1emurY%3D Occurrence Handle10545135
W.R. Leifert C.L. Dorian A. Jahangiri E.J. McMurchie (2001) ArticleTitleDietary fish oil prevents asynchronous contractility and alters Ca2+ handling in adult rat cardiomyocytes. J. Nutr. Biochem. 12 365–376 Occurrence Handle10.1016/S0955-2863(01)00151-6 Occurrence Handle1:CAS:528:DC%2BD3MXksVGrs7w%3D Occurrence Handle11516641
P. Lipp S. Mechmann L. Pott (1987) ArticleTitleEffects of calcium release from sarcoplasmic reticulum on membrane currents in guinea pig atrial cardioballs. Pfluegers Arch. 410 121–131 Occurrence Handle1:STN:280:BieD28jlvVY%3D
P. Lipp L. Pott G. Callewaert E. Carmeliet (1990) ArticleTitleSimultaneous recording of indo-1 fluorescence and Na+/Ca2+ exchange current reveals two components of Ca2+ release from sarcoplasmic reticulum of cardiac atrial myocytes. FEBS Letters 275 181–184 Occurrence Handle10.1016/0014-5793(90)81467-3 Occurrence Handle1:CAS:528:DyaK3MXnvFSqug%3D%3D Occurrence Handle2261988
P.L. McLennan M.Y. Abeywardena J.S. Charnock (1988) ArticleTitleDietary fish oil prevents ventricular fibrillation following coronary artery occlusion and reperfusion. Am. Heart J. 116 709–717 Occurrence Handle1:STN:280:BieA38vgs1Q%3D Occurrence Handle3414486
S.S. Nair J.W. Leitch J. Falconer M.L. Garg (1997) ArticleTitlePrevention of cardiac arrhythmia by dietary (n-3) polyunsaturated fatty acids and their mechanism of action. J. Nutr. 127 383–393 Occurrence Handle1:CAS:528:DyaK2sXhsleisL0%3D Occurrence Handle9082021
N. Negretti M.R. Perez D. Walker S.C. O’Neill (2000) ArticleTitleInhibition of sarcoplasmic reticulum function by polyunsaturated fatty acids in intact, isolated myocytes from rat ventricular muscle. J. Physiol. 523 367–375 Occurrence Handle1:CAS:528:DC%2BD3cXit1yjtLs%3D Occurrence Handle10699081
S.C. O’Neill M.R. Perez K.E. Hammond E.A. Sheader N. Negretti (2002) ArticleTitleDirect and indirect modulation of rat cardiac sarcoplasmic reticulum function by n-3 polyunsaturated fatty acids. J. Physiol. 538 179–184 Occurrence Handle10.1113/jphysiol.2001.013015 Occurrence Handle1:CAS:528:DC%2BD38XislGmsbY%3D Occurrence Handle11773326
E.R. O’Neill M.M. Sakowska D.R. Laver (2003) ArticleTitleRegulation of the calcium release channel from skeletal muscle by suramin and the disulfonated stilbene derivatives DIDS, DBDS & DNDS. Biophys. J. 74 1–16
E.M. Pound J.X. Kang A. Leaf (2001) ArticleTitlePartitioning of polyunsaturated fatty acids, which prevent cardiac arrhythmias, into phospholipid cell membranes. J. Lipid Res. 42 346–351 Occurrence Handle1:CAS:528:DC%2BD3MXisFSrsr8%3D Occurrence Handle11254745
D.A. Saint Y.K. Ju P.W. Gage (1992) ArticleTitleA persistent sodium current in rat ventricular myocytes. J. Physiol. 453 219–231 Occurrence Handle1:STN:280:ByyC3c%2FmsVI%3D Occurrence Handle1334512
F. Striggow B.E. Ehrlich (1997) ArticleTitleRegulation of intracellular calcium release channel function by arachidonic acid and leukotriene B4. Biochim. Biophys. Res. Comm. 237 413–418 Occurrence Handle10.1006/bbrc.1997.7152 Occurrence Handle1:CAS:528:DyaK2sXls1ert74%3D
M. Strokin M. Sergeeva G. Reiser (2003) ArticleTitleDocosahexaenoic acid and arachidonic acid release in rat brain astrocytes is mediated by two separate isoforms of phospholipase A2 and is differently regulated by cyclic AMP and Ca2+. Br. J. Pharmacol. 139 1014–1022 Occurrence Handle10.1038/sj.bjp.0705326 Occurrence Handle1:CAS:528:DC%2BD3sXlvVCisrw%3D Occurrence Handle12839876
G.E. Taffet T.T. Pham D.L. Bick M.L. Entman H.J. Pownall R.J. Bick (1993) ArticleTitleThe calcium uptake of the rat heart sarcoplasmic reticulum is altered by dietary lipid. J. Membrane Biol. 131 35–42 Occurrence Handle1:CAS:528:DyaK3sXht1emsL0%3D
A. Uehara M. Yasukochi I. Imanaga (1996) ArticleTitleModulation of ryanodine binding to the cardiac Ca2+ release channel by arachidonic acid. J. Mol. Cell. Cardiol. 28 43–51 Occurrence Handle10.1006/jmcc.1996.0005 Occurrence Handle1:CAS:528:DyaK28XkvVWqtA%3D%3D Occurrence Handle8745213
W.G. Wier M.B. Cannell J.R. Berlin E. Marban W.J. Lederer (1987) ArticleTitleCellular and subcellular heterogeneity of [Ca2+]i in single heart cells revealed by fura-2. Science 235 325–328 Occurrence Handle1:CAS:528:DyaL2sXot1SjsQ%3D%3D Occurrence Handle3798114
Y.F. Xiao J.X. Kang J.P. Morgan A. Leaf (1995) ArticleTitleBlocking effects of polyunsaturated fatty acids on Na+ channels of neonatal rat ventricular myocytes. Proc. Natl. Acad. Sci. USA 92 11000–11004 Occurrence Handle1:CAS:528:DyaK2MXps1alsrg%3D Occurrence Handle7479925
Y.F. Xiao A.M. Gomez J.P. Morgan W.J. Lederer A. Leaf (1997a) ArticleTitleSuppression of voltage-gated L-type Ca2+ currents by polyunsaturated fatty acids in adult and neonatal rat ventricular myocytes. Proc. Nat. Acad. Sci. USA 94 4182–4187 Occurrence Handle1:CAS:528:DyaK2sXis1amu74%3D
R.P. Xiao H.H. Valdivia K. Bogdanov C. Valdivia E.G. Lakatta H. Cheng (1997b) ArticleTitleThe immunophilin FK506-binding protein modulates Ca2+ release channel closure in rat heart. J. Physiol. 500 343–354 Occurrence Handle1:CAS:528:DyaK2sXjtVWru7s%3D
Acknowledgements
We thank Dr. Manohar Garg for helpful discussions and Melanie Laver for her assistance with the DHA experiments. Also our thanks to Dr. Angela Dulhunty, Suzy Pace and Joan Stivala for supplying SR Vesicles. This work was supported in part by grants from the National Health and Medical Research Council of Australia (to DAS) and the Australian Research Council (to DRL).
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Honen, B., Saint, D. & Laver, D. Suppression of Calcium Sparks in Rat Ventricular Myocytes and Direct Inhibition of Sheep Cardiac RyR Channels by EPA, DHA and Oleic Acid . J. Membrane Biol. 196, 95–103 (2003). https://doi.org/10.1007/s00232-003-0628-9
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DOI: https://doi.org/10.1007/s00232-003-0628-9