Abstract
Neurohumoral stimulation of Gq-coupled receptors has been proposed as a central mechanism in the pathogenesis of diabetic heart disease. The resulting contractile dysfunction is closely related to abnormal intracellular Ca2+ handling with functional defects of the sarcoplasmic reticulum (SR). The present study was therefore designed to determine the role of Gq-protein signaling via Gα11 and Gαq in diabetes for the induction of functional and structural changes in the Ca2+ release complex of the SR. An experimental type 1-diabetes was induced in wild type, Gα11 knockout, and Gα11/q-knockout mice by injection of streptozotocin. Cardiac morphology and function was assessed in vivo by echocardiography. SR Ca2+ leak was tested in vitro based on a 45Ca2+ assay and protein densities as well as gene expression of ryanodine receptor (RyR2), FKBP12.6, sorcin, and annexin A7 were analyzed by immunoblot and RT-PCR. In wild type animals 8 weeks of diabetes resulted in cardiac hypertrophy and SR Ca2+ leak was increased. In addition, diabetic wild type animals showed reduced protein levels of FKBP12.6 and annexin A7. In Gα11- and Gα11/q-knockout animals, however, SR Ca2+ release and cardiac phenotype remained unchanged upon induction of diabetes. Densities of the proteins that we presently analyzed were also unaltered in Gα11-knockout mice. Gα11/q-knockout animals even showed increased expression of sorcin and annexin A7. Thus, based on the present study we suggest a signaling pathway via the Gq-proteins, Gα11 and Gαq, that could link increased neurohumoral stimulation in diabetes with defective RyR2 channel function by regulating protein expression of FKBP12.6, annexin A7, and sorcin.
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References
Riggs TW, Transue D (1990) Doppler echocardiographic evaluation of left ventricular diastolic function in adolescents with diabetes mellitus. Am J Cardiol 65:899–902
Lababidi ZA, Goldstein DE (1983) High prevalence of echocardiographic abnormalities in diabetic youths. Diabetes Care 6:18–22
Belke DD, Dillmann WH (2004) Altered cardiac calcium handling in diabetes. Curr Hypertens Rep 6:424–429
Reuter H, Gronke S, Adam C, Ribati M, Brabender J, Zobel C, Frank KF, Wippermann J, Schwinger RH, Brixius K, Muller-Ehmsen J (2008) Sarcoplasmic Ca(2+) release is prolonged in nonfailing myocardium of diabetic patients. Mol Cell Biochem 308:141–149
Teshima Y, Takahashi N, Saikawa T, Hara M, Yasunaga S, Hidaka S, Sakata T (2000) Diminished expression of sarcoplasmic reticulum Ca(2+)-ATPase and ryanodine sensitive Ca(2+)Channel mRNA in streptozotocin-induced diabetic rat heart. J Mol Cell Cardiol 32:655–664
Farrell EF, Antaramian A, Rueda A, Gomez AM, Valdivia HH (2003) Sorcin inhibits calcium release and modulates excitation-contraction coupling in the heart. J Biol Chem 278:34660–34666
Herr C, Smyth N, Ullrich S, Yun F, Sasse P, Hescheler J, Fleischmann B, Lasek K, Brixius K, Schwinger RH, Fassler R, Schroder R, Noegel AA (2001) Loss of annexin A7 leads to alterations in frequency-induced shortening of isolated murine cardiomyocytes. Mol Cell Biol 21:4119–4128
Wehrens XH, Lehnart SE, Marks AR (2005) Intracellular calcium release and cardiac disease. Annu Rev Physiol 67:69–98
Marx SO, Reiken S, Hisamatsu Y, Jayaraman T, Burkhoff D, Rosemblit N, Marks AR (2000) PKA phosphorylation dissociates FKBP12.6 from the calcium release channel (ryanodine receptor): defective regulation in failing hearts. Cell 101:365–376
Yaras N, Ugur M, Ozdemir S, Gurdal H, Purali N, Lacampagne A, Vassort G, Turan B (2005) Effects of diabetes on ryanodine receptor Ca release channel (RyR2) and Ca2+ homeostasis in rat heart. Diabetes 54:3082–3088
Suarez J, Belke DD, Gloss B, Dieterle T, McDonough PM, Kim YK, Brunton LL, Dillmann WH (2004) In vivo adenoviral transfer of sorcin reverses cardiac contractile abnormalities of diabetic cardiomyopathy. Am J Physiol Heart Circ Physiol 286:H68–H75
Bidasee KR, Nallani K, Yu Y, Cocklin RR, Zhang Y, Wang M, Dincer UD, Besch HR Jr (2003) Chronic diabetes increases advanced glycation end products on cardiac ryanodine receptors/calcium-release channels. Diabetes 52:1825–1836
Dhalla NS, Liu X, Panagia V, Takeda N (1998) Subcellular remodeling and heart dysfunction in chronic diabetes. Cardiovasc Res 40:239–247
Reuter H, Adam C, Gronke S, Zobel C, Frank KF, Muller-Ehmsen J, Brabender J, Schwinger RH (2006) The increased angiotensin II (type 1) receptor density in myocardium of type 2 diabetic patients is prevented by blockade of the renin-angiotensin system. Diabetologia 49:3067–3074
Koya D, King GL (1998) Protein kinase C activation and the development of diabetic complications. Diabetes 47:859–866
Eurich DT, Majumdar SR, Tsuyuki RT, Johnson JA (2004) Reduced mortality associated with the use of ACE inhibitors in patients with type 2 diabetes. Diabetes Care 27:1330–1334
Lindholm LH, Ibsen H, Dahlof B, Devereux RB, Beevers G, de Faire U, Fyhrquist F, Julius S, Kjeldsen SE, Kristiansson K, Lederballe-Pedersen O, Nieminen MS, Omvik P, Oparil S, Wedel H, Aurup P, Edelman J, Snapinn S (2002) Cardiovascular morbidity and mortality in patients with diabetes in the losartan intervention for endpoint reduction in hypertension study (LIFE): a randomised trial against atenolol. Lancet 359:1004–1010
Ozdemir S, Ugur M, Gurdal H, Turan B (2005) Treatment with AT(1) receptor blocker restores diabetes-induced alterations in intracellular Ca(2+) transients and contractile function of rat myocardium. Arch Biochem Biophys 435:166–174
Wolkart G, Pang X, Stessel H, Kirchengast M, Brunner F (2007) Chronic endothelin-A receptor antagonism is as protective as angiotensin converting enzyme inhibition against cardiac dysfunction in diabetic rats. Br J Pharmacol 151:1187–1197
Offermanns S, Zhao LP, Gohla A, Sarosi I, Simon MI, Wilkie TM (1998) Embryonic cardiomyocyte hypoplasia and craniofacial defects in G alpha q/G alpha 11-mutant mice. EMBO J 17:4304–4312
Wettschureck N, Rutten H, Zywietz A, Gehring D, Wilkie TM, Chen J, Chien KR, Offermanns S (2001) Absence of pressure overload induced myocardial hypertrophy after conditional inactivation of Galphaq/Galpha11 in cardiomyocytes. Nat Med 7:1236–1240
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986–994
Meissner G, Henderson JS (1987) Rapid calcium release from cardiac sarcoplasmic reticulum vesicles is dependent on Ca2+ and is modulated by Mg2+, adenine nucleotide, and calmodulin. J Biol Chem 262:3065–3073
Meyer M, Schillinger W, Pieske B, Holubarsch C, Heilmann C, Posival H, Kuwajima G, Mikoshiba K, Just H, Hasenfuss G et al (1995) Alterations of sarcoplasmic reticulum proteins in failing human dilated cardiomyopathy. Circulation 92:778–784
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Luo W, Grupp IL, Harrer J, Ponniah S, Grupp G, Duffy JJ, Doetschman T, Kranias EG (1994) Targeted ablation of the phospholamban gene is associated with markedly enhanced myocardial contractility and loss of beta-agonist stimulation. Circ Res 75:401–409
Bidasee KR, Nallani K, Besch HR Jr, Dincer UD (2003) Streptozotocin-induced diabetes increases disulfide bond formation on cardiac ryanodine receptor (RyR2). J Pharmacol Exp Ther 305:989–998
Brillantes AB, Ondrias K, Scott A, Kobrinsky E, Ondriasova E, Moschella MC, Jayaraman T, Landers M, Ehrlich BE, Marks AR (1994) Stabilization of calcium release channel (ryanodine receptor) function by FK506-binding protein. Cell 77:513–523
Marx SO, Ondrias K, Marks AR (1998) Coupled gating between individual skeletal muscle Ca2+ release channels (ryanodine receptors). Science 281:818–821
Shao CH, Rozanski GJ, Patel KP, Bidasee KR (2007) Dyssynchronous (non-uniform) Ca2+ release in myocytes from streptozotocin-induced diabetic rats. J Mol Cell Cardiol 42:234–246
Lehnart SE, Terrenoire C, Reiken S, Wehrens XH, Song LS, Tillman EJ, Mancarella S, Coromilas J, Lederer WJ, Kass RS, Marks AR (2006) Stabilization of cardiac ryanodine receptor prevents intracellular calcium leak and arrhythmias. Proc Natl Acad Sci USA 103:7906–7910
Stange M, Xu L, Balshaw D, Yamaguchi N, Meissner G (2003) Characterization of recombinant skeletal muscle (Ser-2843) and cardiac muscle (Ser-2809) ryanodine receptor phosphorylation mutants. J Biol Chem 278:51693–51702
Galderisi M, Anderson KM, Wilson PW, Levy D (1991) Echocardiographic evidence for the existence of a distinct diabetic cardiomyopathy (the Framingham Heart Study). Am J Cardiol 68:85–89
Dolmetsch RE, Lewis RS, Goodnow CC, Healy JI (1997) Differential activation of transcription factors induced by Ca2+ response amplitude and duration. Nature 386:855–858
Acknowledgments
Our special thanks go to Prof. Dr. A. Nögel (Institute of biochemistry of the University of Cologne) for providing the annexin A7 antibody. This study was supported by the German Heart Foundation/German Foundation of Heart Research (F/11/07 to H. Reuter). This work is part of the dissertation of D. Hoyer (University of Cologne, in preparation).
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Hoyer, D.P., Grönke, S., Frank, K.F. et al. Diabetes-related defects in sarcoplasmic Ca2+ release are prevented by inactivation of Gα11 and Gαq in murine cardiomyocytes. Mol Cell Biochem 341, 235–244 (2010). https://doi.org/10.1007/s11010-010-0454-1
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DOI: https://doi.org/10.1007/s11010-010-0454-1