Pflügers Archiv - European Journal of Physiology

, Volume 465, Issue 10, pp 1477–1486 | Cite as

Transgenic rescue of defective Cd36 enhances myocardial adenylyl cyclase signaling in spontaneously hypertensive rats

  • Martina Klevstig
  • Dmitry Manakov
  • Dita Kasparova
  • Iveta Brabcova
  • Frantisek Papousek
  • Jitka Zurmanova
  • Vaclav Zidek
  • Jan Silhavy
  • Jan Neckar
  • Michal Pravenec
  • Frantisek Kolar
  • Olga Novakova
  • Jiri NovotnyEmail author
Organ physiology


Dysfunction or abnormalities in the regulation of fatty acid translocase Cd36, a multifunctional membrane protein participating in uptake of long-chain fatty acids, has been linked to the development of heart diseases both in animals and humans. We have previously shown that the Cd36 transgenic spontaneously hypertensive rat (SHR-Cd36), with a wild type Cd36, has higher susceptibility to ischemic ventricular arrhythmias when compared to spontaneously hypertensive rat (SHR) carrying a mutant Cd36 gene, which may have been related to increased β-adrenergic responsiveness of these animals (Neckar et al., 2012 Physiol. Genomics 44:173–182). The present study aimed to determine whether the insertion of the wild type Cd36 into SHR would affect the function of myocardial G protein-regulated adenylyl cyclase (AC) signaling. β-Adrenergic receptors (β-ARs) were characterized by radioligand-binding experiments and the expression of selected G protein subunits, AC, and protein kinase A (PKA) was determined by RT-PCR and Western blot analyses. There was no significant difference in the amount of trimeric G proteins, but the number of β-ARs was higher (by about 35 %) in myocardial preparations from SHR-Cd36 as compared to SHR. Besides that, transgenic rats expressed increased amount (by about 20 %) of the dominant myocardial isoforms AC5/6 and contained higher levels of both nonphosphorylated (by 11 %) and phosphorylated (by 45 %) PKA. Differently stimulated AC activity in SHR-Cd36 significantly exceeded (by about 18–30 %) the enzyme activity in SHR. Changes at the molecular level were reflected by higher contractile responses to stimulation by the adrenergic agonist dobutamine. In summary, it can be concluded that the increased susceptibility to ischemic arrhythmias of SHR-Cd36 is attributable to upregulation of some components of the β-AR signaling pathway, which leads to enhanced sensitization of AC and increased cardiac adrenergic responsiveness.


SHR rats Cd36 Heart β-Adrenergic receptors Adenylyl cyclase Protein kinase A 



This work was supported by the Charles University Grant Agency (429611); the Ministry of Education, Youth, and Sports of the Czech Republic (MSM0021620858 and LL1204 within the ERC CZ program); the Grant Agency of the Academy of Science of the Czech Republic (IAAX01110901, IAA601110908, and P303/10/0505); and by the grant SVV-2012-265211.


  1. 1.
    Aitman TJ, Gotoda T, Evans AL, Imrie H, Heath KE, Trembling PM, Truman H, Wallace CA, Rahman A, Dore C, Flint J, Kren V, Zidek V, Kurtz TW, Pravenec M, Scott J (1997) Quantitative trait loci for cellular defects in glucose and fatty acid metabolism in hypertensive rats. Nat Genet 16:197–201PubMedCrossRefGoogle Scholar
  2. 2.
    Anand-Srivastava MB (1988) Altered responsiveness of adenylate-cyclase to adenosine and other agents in the myocardial sarcolemma and aorta of spontaneously-hypertensive rats. Biochem Pharmacol 37:3017–3022PubMedCrossRefGoogle Scholar
  3. 3.
    Antos CL, Frey N, Marx SO, Reiken S, Gaburjakova M, Richardson JA, Marks AR, Olson EN (2001) Dilated cardiomyopathy and sudden death resulting from constitutive activation of protein kinase A. Circ Res 89:997–1004PubMedCrossRefGoogle Scholar
  4. 4.
    Bassiakou E, Xanthos T, Papadimitriou L (2009) The potential beneficial effects of beta adrenergic blockade in the treatment of ventricular fibrillation. Eur J Pharmacol 616:1–6PubMedCrossRefGoogle Scholar
  5. 5.
    Bernstein D, Fajardo G, Zhao M (2011) The role of beta-adrenergic receptors in heart failure: differential regulation of cardiotoxicity and cardioprotection. Prog Pediat Cardiol 31:35–38CrossRefGoogle Scholar
  6. 6.
    Bohuslavova R, Kolar F, Kuthanova L, Neckar J, Tichopad A, Pavlinkova G (2010) Gene expression profiling of sex differences in HIF1-dependent adaptive cardiac responses to chronic hypoxia. J Appl Physiol 109:1195–1202PubMedCrossRefGoogle Scholar
  7. 7.
    Bonen A, Han XX, Tandon NN, Glatz JF, Lally J, Snook LA, Luiken JJ (2009) FAT/CD36 expression is not ablated in spontaneously hypertensive rats. J Lipid Res 50:740–748PubMedCrossRefGoogle Scholar
  8. 8.
    Dai WD, Simkhovich BZ, Kloner RA (2009) Ischemic preconditioning maintains cardioprotection in aging normotensive and spontaneously hypertensive rats. Exp Gerontol 44:344–349PubMedCrossRefGoogle Scholar
  9. 9.
    Devic E, Xiang Y, Gould D, Kobilka B (2001) Beta-adrenergic receptor subtype-specific signaling in cardiac myocytes from beta(1) and beta(2) adrenoceptor knockout mice. Mol Pharmacol 60:577–583PubMedGoogle Scholar
  10. 10.
    Di Benedetto G, Zoccarato A, Lissandron V, Terrin A, Li X, Houslay MD, Baillie GS, Zaccolo M (2008) Protein kinase a type I and type II define distinct intracellular signaling compartments. Circ Res 103:836–844PubMedCrossRefGoogle Scholar
  11. 11.
    El-Armouche A, Eschenhagen T (2009) Beta-adrenergic stimulation and myocardial function in the failing heart. Heart Fail Rev 14:225–241PubMedCrossRefGoogle Scholar
  12. 12.
    Engelhardt S, Hein L, Wiesmann F, Lohse MJ (1999) Progressive hypertrophy and heart failure in beta(1)-adrenergic receptor transgenic mice. Proc Natl Acad Scie USA 96:7059–7064CrossRefGoogle Scholar
  13. 13.
    Frolikova M, Stopkova R, Antalikova J, Johnson PM, Stopka P, Dvorakova-Hortova K (2012) Role of complement regulatory proteins CD46, CD55 and CD59 in reproduction. Folia Zool 61:84–94Google Scholar
  14. 14.
    Girouard H, Chulak C, LeJossec M, Lamontagne D, de Champlain J (2003) Chronic antioxidant treatment improves sympathetic functions and beta-adrenergic pathway in the spontaneously hypertensive rats. J Hypertens 21:179–188PubMedCrossRefGoogle Scholar
  15. 15.
    Hajri T, Ibrahimi A, Coburn CT, Knapp FF, Kurtz T, Pravenec M, Abumrad NA (2001) Defective fatty acid uptake in the spontaneously hypertensive rat is a primary determinant of altered glucose metabolism, hyperinsulinemia, and myocardial hypertrophy. J Biol Chem 276:23661–23666PubMedCrossRefGoogle Scholar
  16. 16.
    Hall D, Mayosi BM, Rahman TJ, Avery PJ, Watkins HC, Keavney B (2011) Common variation in the CD36 (fatty acid translocase) gene is associated with left-ventricular mass. J Hypertens 29:690–695PubMedCrossRefGoogle Scholar
  17. 17.
    Irie H, Krukenkamp IB, Brinkmann JF, Gaudette GR, Saltman AE, Jou W, Glatz JF, Abumrad NA, Ibrahimi A (2003) Myocardial recovery from ischemia is impaired in CD36-null mice and restored by myocyte CD36 expression or medium-chain fatty acids. Proc Natl Acad Sci USA 100:6819–6824PubMedCrossRefGoogle Scholar
  18. 18.
    Klevstig MJ, Markova I, Burianova J, Kazdova L, Pravenec M, Novakova O, Novak F (2011) Role of FAT/CD36 in novel PKC isoform activation in heart of spontaneously hypertensive rats. Mol Cell Biochem 357:163–169PubMedCrossRefGoogle Scholar
  19. 19.
    Kuang M, Febbraio M, Wagg C, Lopaschuk GD, Dyck JRB (2004) Fatty acid translocase/CD36 deficiency does not energetically or functionally compromise hearts before or after ischemia. Circulation 109:1550–1557PubMedCrossRefGoogle Scholar
  20. 20.
    Leenen FHH, Yuan BX (2001) Mortality after coronary artery occlusion in different models of cardiac hypertrophy in rats. Hypertension 37:209–215PubMedCrossRefGoogle Scholar
  21. 21.
    Liggett SB, Tepe NM, Lorenz JN, Canning AM, Jantz TD, Mitarai S, Yatani A, Dorn GW (2000) Early and delayed consequences of beta(2)-adrenergic receptor overexpression in mouse hearts—critical role for expression level. Circulation 101:1707–1714PubMedCrossRefGoogle Scholar
  22. 22.
    Lissandron V, Zaccolo M (2006) Compartmentalized cAMP/PKA signalling regulates cardiac excitation–contraction coupling. J Muscle Res Cell Motil 27:399–403PubMedCrossRefGoogle Scholar
  23. 23.
    Lu YJ, Zhang Y, Shan HL, Pan ZW, Li XL, Li BX, Xu CQ, Zhang BS, Zhang FM, Dong DL, Song WQ, Qiao GF, Yang BF (2009) MicroRNA-1 downregulation by propranolol in a rat model of myocardial infarction: a new mechanism for ischaemic cardioprotection. Cardiovasc Res 84:434–441PubMedCrossRefGoogle Scholar
  24. 24.
    Matsumori Y, Ohyanagi M, Kawamoto H, Shibata R, Iwasaki T (1989) Intracellular distribution of cardiac beta-adrenoceptors in SHR and WKY. Jpn Circ J 53:113–120PubMedCrossRefGoogle Scholar
  25. 25.
    Neckar J, Silhavy J, Zidek V, Landa V, Mlejnek P, Simakova M, Seidman JG, Seidman C, Kazdova L, Klevstig M, Novak F, Vecka M, Papousek F, Houstek J, Drahota Z, Kurtz TW, Kolar F, Pravenec M (2012) CD36 overexpression predisposes to arrhythmias but reduces infarct size in spontaneously hypertensive rats: gene expression profile analysis. Physiol Genomics 44:173–182PubMedCrossRefGoogle Scholar
  26. 26.
    Okumura S, Vatner DE, Kurotani R, Bai Y, Gao SM, Yuan ZR, Iwatsubo K, Ulucan C, Kawabe J, Ghosh K, Vatner SF, Ishikawa Y (2007) Disruption of type 5 adenylyl cyclase enhances desensitization of cyclic adenosine monophosphate signal and increases Akt signal with chronic catecholamine stress. Circulation 116:1776–1783PubMedCrossRefGoogle Scholar
  27. 27.
    Penna C, Tullio F, Moro F, Folino A, Merlino A, Pagliaro P (2010) Effects of a protocol of ischemic postconditioning and/or captopril in hearts of normotensive and hypertensive rats. Basic Res Cardiol 105:181–192PubMedCrossRefGoogle Scholar
  28. 28.
    Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45PubMedCrossRefGoogle Scholar
  29. 29.
    Pietka TA, Sulkin MS, Kuda O, Wang W, Zhou D, Yamada KA, Yang K, Su X, Gross RW, Nerbonne JM, Efimov IR, Abumrad NA (2012) CD36 influences myocardial Ca2+ homeostasis and phospholipid metabolism: conduction anomalies in CD36 deficient mice during fasting. JBiol Chem 287:38901–38912CrossRefGoogle Scholar
  30. 30.
    Pravenec M, Gauguier D, Schott JJ, Buard J, Kren V, Bila V, Szpirer C, Szpirer J, Wang JM, Huang HM, Stlezin E, Spence MA, Flodman P, Printz M, Lathrop GM, Vergnaud G, Kurtz TW (1995) Mapping of quantitative trait loci for blood pressure and cardiac mass in the rat by genome scanning of recombinant inbred strains. J Clin Invest 96:1973–1978PubMedCrossRefGoogle Scholar
  31. 31.
    Pravenec M, Landa V, Zidek V, Musilova A, Kren V, Kazdova L, Aitman TJ, Glazier AM, Ibrahimi A, Abumrad NA, Qi NN, Wang JM, St Lezin EM, Kurtz TW (2001) Transgenic rescue of defective Cd36 ameliorates insulin resistance in spontaneously hypertensive rats. Nat Genet 27:156–158PubMedCrossRefGoogle Scholar
  32. 32.
    Ravingerova T, Bernatova I, Matejikova J, Ledvenyiova V, Nemcekova M, Pechanova O, Tribulova N, Slezak J (2011) Impaired cardiac ischemic tolerance in spontaneously hypertensive rats is attenuated by adaptation to chronic and acute stress. Exp Clin Cardiol 16:E23–E29PubMedGoogle Scholar
  33. 33.
    Roberts W, Magwenzi S, Aburima A, Naseem KM (2010) Thrombospondin-1 induces platelet activation through CD36-dependent inhibition of the cAMP/protein kinase A signaling cascade. Blood 116:4297–4306PubMedCrossRefGoogle Scholar
  34. 34.
    Slotkin TA, Auman JT, Seidler FJ (2003) Ontogenesis of beta-adrenoceptor signaling: implications for perinatal physiology and for fetal effects of tocolytic drugs. J PharmacolExp Ther 306:1–7CrossRefGoogle Scholar
  35. 35.
    Soto D, De Arcangelis V, Zhang J, Xiang Y (2009) Dynamic protein kinase A activities induced by beta-adrenoceptors dictate signaling propagation for substrate phosphorylation and myocyte contraction. Circ Res 104:770–U121PubMedCrossRefGoogle Scholar
  36. 36.
    Sung MMY, Koonen DPY, Soltys CLM, Jacobs RL, Febbraio M, Dyck JRB (2011) Increased CD36 expression in middle-aged mice contributes to obesity-related cardiac hypertrophy in the absence of cardiac dysfunction. J Mol Med 89:459–469PubMedCrossRefGoogle Scholar
  37. 37.
    Takahashi T, Tang T, Lai NC, Roth DM, Rebolledo B, Saito M, Lew WYW, Clopton P, Hammond HK (2006) Increased cardiac adenylyl cyclase expression is associated with increased survival after myocardial infarction. Circulation 114:388–396PubMedCrossRefGoogle Scholar
  38. 38.
    Takata Y, Kato H (1995) Adrenoceptors in SHR—alterations in binding characteristics and intracellular signal-transduction pathways. Life Sci 58:91–106CrossRefGoogle Scholar
  39. 39.
    Tang T, Gao MH, Lai NC, Firth AL, Takahashi T, Guo T, Yuan JXJ, Roth DM, Hammond HK (2008) Adenylyl cyclase type 6 deletion decreases left ventricular function via impaired calcium handling. Circulation 117:61–69PubMedCrossRefGoogle Scholar
  40. 40.
    Xiang Y, Kobilka BK (2003) Myocyte adrenoceptor signaling pathways. Science 300:1530–1532PubMedCrossRefGoogle Scholar
  41. 41.
    Xiao RP, Zhu WZ, Zheng M, Cao CM, Zhang YY, Lakatta EG, Han Q (2006) Subtype-specific alpha(1)- and beta-adrenoceptor signaling in the heart. Trends Pharmacol Sci 27:330–337PubMedCrossRefGoogle Scholar
  42. 42.
    Zhang L, Xu CQ, Hong YA, Zhang JL, Liu Y, Zhao M, Cao YX, Lu YJ, Yang BF, Shan HL (2010) Propranolol regulates cardiac transient outward potassium channel in rat myocardium via cAMP/PKA after short-term but not after long-term ischemia. Naunyn-Schmiedebergs Arch Pharmacol 382:63–71PubMedCrossRefGoogle Scholar
  43. 43.
    Zhu WZ, Zheng M, Koch WJ, Lefkowitz RJ, Kobilka BK, Xiao RP (2001) Dual modulation of cell survival and cell death by beta(2)-adrenergic signaling in adult mouse cardiac myocytes. Proc Natl Acad SciUSA 98:1607–1612CrossRefGoogle Scholar
  44. 44.
    Zolk O, Kouchi I, Schnabel P, Bohm M (2000) Heterotrimeric G proteins in heart disease. Can JPhysiol Pharmacol 78:187–198CrossRefGoogle Scholar
  45. 45.
    Zurmanova J, Puta F, Stopkova T, Soukup T (2008) Real time RT-PCR with a newly designed set of primers confirmed the presence of 2b and 2x/d myosin heavy chain mRNAs in the rat slow soleus muscle. Physiol Res 57:973–978PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Martina Klevstig
    • 1
  • Dmitry Manakov
    • 2
  • Dita Kasparova
    • 2
  • Iveta Brabcova
    • 2
  • Frantisek Papousek
    • 3
  • Jitka Zurmanova
    • 2
  • Vaclav Zidek
    • 3
  • Jan Silhavy
    • 3
  • Jan Neckar
    • 3
  • Michal Pravenec
    • 3
  • Frantisek Kolar
    • 3
  • Olga Novakova
    • 1
  • Jiri Novotny
    • 2
    Email author
  1. 1.Department of Cell Biology, Faculty of ScienceCharles University in PraguePrague 2Czech Republic
  2. 2.Department of Physiology, Faculty of ScienceCharles University in PraguePrague 2Czech Republic
  3. 3.Institute of PhysiologyAcademy of Sciences of the Czech RepublicPragueCzech Republic

Personalised recommendations