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Regulation of ß-adrenoceptors in the guinea-pig sinoatrial node

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Abstract

This study examined the changes of ß-adrenoceptors in the guinea-pig sinoatrial nodal region following 7 day (−)-isoprenaline (400 μg/kg/h s.c.) infusion and the relationship between ß-adrenoceptor desensitization and receptor down-regulation. Changes in ß1-and ß2-adrenoceptor density were measured using quantitative autoradiography and function in organ bath studies.

(−)-Isoprenaline treatment produced a marked decrease in total (from 57.5 to 33.9 fmol/mg protein), ß1-(from 49.4 to 32.8 fmol/mg protein), and ß2-adrenoceptor density (from 8.1 to 1.05 fmol/mg protein) in the sinoatrial node. In adjacent right atrium, treatment produced no change in total (39.5 and 36.7 fmol/mg protein) or ß1-adrenoceptors (35.9 and 36.4 fmol/mg protein) but did decrease ß2-adrenoceptors (from 3.7 to 0.3 fmol/mg protein). Chronotropic effects were measured in spontaneously beating right atrium. Procaterol, a selective ß2-adrenoceptor agonist, caused a biphasic chronotropic response in control right atria, the first part of which was abolished in the tissue from treated animals. The maximum increase in right atrial rate to R0363, a ß1-adrenoceptor selective partial agonist, was reduced from 114 bpm in control to 43 bpm in treated animals. In electrically driven right atrium with the sinoatrial node removed procaterol failed to produce a positive inotropic response via ß2-adrenoceptors, but the maximum response to RO 363 was reduced from 0.75 g in the control tissue to 0.12 g in the treated tissue.

This study showed that changes in ß2-adrenoceptor density following 7 day (−)-isoprenaline infusion are compatible with reduced functional responsiveness in the SA node. The reduction of al-adrenoceptor number in the SA node was also compatible with the reduced chronotropic response in this tissue. However the lack of effect on ß1-adrenoceptor density in the right atrium was not consistent with the decrease in Qt-adrenoceptor mediated inotropic response in this tissue. This suggests that ß-adrenoceptor desensitization is not always associated with receptor down-regulation but depends also on the changes in the cell signalling system beyond the level of the receptor which differ according to the cardiac location.

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References

  • Benovic JL, Strasser RH, Caron MG, Lefkowitz RJ (1986) ß-Adrenergic receptor kinase: identification of a novel protein kinase that phosphorylates the agonist-occupied form of the receptor. Proc Natl Acad Sci USA 83:2797–2801

    Article  PubMed  CAS  Google Scholar 

  • Bilski AJ, Halliday SE, Fitzgerald JD, Wale JL (1983) The pharmacology of a ß2-selective adrenoceptor antagonist (ICI 118,551). J Cardiovasc Pharmacol 5:430–437

    Article  PubMed  CAS  Google Scholar 

  • Boullin DJ, Costa E, Brodie BB (1967) Evidence that blockade of adrenergic receptors causes overflow of norepinephrine in cat's colon after nerve stimulation. J Pharmacol Exp Ther 157:125–134

    PubMed  CAS  Google Scholar 

  • Bouvier M, Collins S, O'Dowd BF, Campbell PT, de Blasi A, Kobilka BK, MacGregor C, Irons GP, Caron MG, Lefkowitz RJ (1989) Two distinct pathways for CAMP-mediated down-regulation of the ß2-adrenergic receptor. J Biol Chem 264:16786–16792

    PubMed  CAS  Google Scholar 

  • Brown JE, McLeod AA, Shand DG (1986) In support of cardiac chronotropic beta2 adrenoceptors. Am J Cardiol 57:1117–1617

    Article  Google Scholar 

  • Bull G, Molenaar P, Summers RJ (1990) Functional and receptor binding studies in guinea-pig heart after acute and chronic (-)isoprenaline infusion [Abstract]. Clin Exp Pharmacol Physiol 16(suppl):133

    Google Scholar 

  • Buxton BF, Jones CR, Molenaar P, Summers RJ (1987) Characterization and autoradiographic localization of ß-adrenoceptor subtypes in human cardiac tissues. Br J Pharmacol 92:299–310

    PubMed  CAS  Google Scholar 

  • Cabral AN, Vasquez EC (1984) Cardiac beta-adrenoceptor desensitization after sinoaortic baroreceptors denervation or isoproterenol-pretreatment. Pharmacol Res Commun 16:1031–1040

    Article  PubMed  CAS  Google Scholar 

  • Chuang D-M, Costa E (1979) Evidence for internalization of the recognition site of ß-adrenergic receptors during receptor subsensitivity induced by (−)-isoproterenol. Proc Natl Acad Sci USA 76: 3024–3028

    Article  PubMed  CAS  Google Scholar 

  • Clark RB, Friedman J, Dixon RAF, Strader CD (1989) Identification of a specific site required for rapid heterologous desensitization of the ß-adrenergic receptor by cAMP-dependent protein kinase. Mol Pharmacol 36:343–348

    PubMed  CAS  Google Scholar 

  • Desaulles E, Miesch F, Schwartz J (1978) Evidence for the participation of ß1-adrenoceptors in isoprenaline-induced renin release from rat kidney slices in vitro. Br J Pharmacol 63:421–425

    PubMed  CAS  Google Scholar 

  • Dooley DJ, Bittiger H, Reyman NC (1986) CGP 20712 A: a useful tool for quantitating ß1- and ß2-adrenoceptors. Eur J Pharmacol 130:137–140

    Article  PubMed  CAS  Google Scholar 

  • Eschenhagen T, Mende U, Nose M, Schmitz W, Scholz H, Warnholtz A, Wustel J-M (1991) Isoprenaline-induced increase in mRNA levels of inhibitory G-protein α-subunits in rat heart. Naunyn-Schmiedebergs Arch Pharmacol 343:609–615

    PubMed  CAS  Google Scholar 

  • Gambarana C, Ordway GA, Hauptmann M, Tejani-Butt S, Frazer A (1991) Central administration of l-isoproterenol in vivo induces a preferential regulation of ß2-adrenoceptors in the central nervous system of the rat. Brain Res 555:141–148

    Article  PubMed  CAS  Google Scholar 

  • Hadjiivanova N, Flint N, Evans WH, Dix C, Cooke BA (1984) Endocytosis of ß-adrenergic ligands by rat liver. Comparison of ß-adrenergic receptor and adenylate cyclase distribution in endosome and plasma-membrane fractions. Biochem J 222:749–754

    PubMed  CAS  Google Scholar 

  • Harden TK (1983) Agonist-induced desensitization of the ß-adrenergic receptor-linked adenylate cyclase. Pharmacol Rev 35:5–32

    PubMed  CAS  Google Scholar 

  • Hausdorff WP, Caron MG, Lefkowitz RJ (1990) Turning off the signal: desensitization of ß-adrenergic receptor function. FASEB J 4:2881–2889

    PubMed  CAS  Google Scholar 

  • Hawthorn MH and Broadley KJ (1982) ß-Adrenoceptor ligand binding and supersensitivity to isoprenaline of ventricular muscle after chronic reserpine pretreatment. Naunyn-Schmiedebergs Arch Pharmacol 320:240–245

    Article  PubMed  CAS  Google Scholar 

  • Hertel C, Staehelin M (1983) Reappearance of ß-adrenergic receptors after isoprenaline treatment in intact C6-cells. J Cell Biol 97: 1538–1543

    Article  PubMed  CAS  Google Scholar 

  • Hertel C, Staehelin M, Perkins JP (1983) Evidence for intravesicular ß-adrenergic receptors in membrane fractions from desensitized cells: binding of the hydrophilic ligand CGP-12177 only in the presence of alamethicin. J Cyclic Nucleotide Protein Phosphor Res 9: 119–128

    PubMed  CAS  Google Scholar 

  • Johansson L-H, Persson H (1983) ß2-Adrenoceptors in guinea-pig atria. J Pharm Pharmacol 35:804–807

    PubMed  CAS  Google Scholar 

  • Kaumann AJ, Morris TH, Bojar H (1983) Heart ß-receptors: on the functional role of heterogeneous binding sites. J Recept Res 3:61–70

    PubMed  CAS  Google Scholar 

  • Kaumann AJ (1986) The ß1-adrenoceptor antagonist CGP 20712A unmasks ß2-adrenoceptors activated by (−)-adrenaline in rat sinoatrial node. Naunyn-Schmiedebergs Arch Pharmacol 332:406–409

    Article  PubMed  CAS  Google Scholar 

  • Kaumann AJ, Lemoine H (1985) Direct labelling of myocardial ß1-adrenoceptors. Comparison of binding affinity of 3H-(−)-bisoprolol with its blocking potency. Naunyn-Schmiedebergs Arch Pharmacol 331:27–39

    Article  PubMed  CAS  Google Scholar 

  • Kaumann AJ, Lobnig BM (1986) Mode of action of (−)-pindolol on feline and human myocardium. Br J Pharmacol 89:207–218

    PubMed  CAS  Google Scholar 

  • Kazanietz MG, Enero MA (1989) Modulation of noradrenaline release by presynaptic alpha-2 and beta adrenoceptors in rat atria. Naunyn-Schmiedebergs Arch Pharmacol 340:274–278

    PubMed  CAS  Google Scholar 

  • Kazanietz MG, Enero MA (1992) Role of cyclic AMP in the release of noradrenaline from isolated rat atria: effect of pretreatment with clenbuterol. Naunyn-Schmiedebergs Arch Pharmacol 346:311–314

    PubMed  CAS  Google Scholar 

  • Latifpour J and McNeill JH (1984) Reserpine-induced changes in cardiac adrenergic receptors. Can J Physiol Pharmacol 62:23–26

    PubMed  CAS  Google Scholar 

  • Lemoine H, Ehle B, Kaumann AJ (1985) Direct labelling of ß2-adrenoceptors. Comparison of binding potency of 3H-ICI 118,551 and blocking potency of IC 118,551. Naunyn-Schmiedebergs Arch Pharmacol 331:40–51

    Article  PubMed  CAS  Google Scholar 

  • Maisel AS, Phillips C, Michel MC, Ziegler MG, Carter SM (1989) Regulation of cardiac ß-adrenergic receptors by captopril: implications for congestive heart failure. Circulation 80:669–675

    PubMed  CAS  Google Scholar 

  • Majewski H (1983) Modulation of noradrenaline release through activation of presynaptic ß-adrenoceptors. J Anton Pharmacol 3:47–60

    Article  CAS  Google Scholar 

  • Majewski H (1989) Angiotensin II and noradrenergic transmission in the pithed rat. J Cardiovasc Pharmacol 14:622–630

    Article  PubMed  CAS  Google Scholar 

  • McGonigle P, Neve KA, Molinoff PB (1986) A quantitative method of analyzing the interaction of slightly selective radioligands with multiple receptor subtypes. Mol Pharmacol 30:329–337

    PubMed  CAS  Google Scholar 

  • McPherson GA, Malta E, Molenaar P, Raper C (1984) The affinity and efficacy of the selective ß1-adrenoceptor stimulant R0363 ß1-and ß2-adrenoceptor binding sites. Br J Pharmacol 82:897–904

    PubMed  CAS  Google Scholar 

  • Mende U, Eschenhagen T, Geertz B, Schmitz W, Scholz H, Esch JS, Sempell R, Steinfath M (1992) Isoprenaline-induced increase in the 40/41 kDa pertussis toxin substrates and functional consequences on contractile response in rat heart. Naunyn-Schmiedbergs Arch Pharmacol 345:44–50

    CAS  Google Scholar 

  • Mickey JV, Tate R, Mullikin D, Lefkowitz RJ (1976) Regulation of adenylate cyclase-coupled beta adrenergic receptor binding sites by beta adrenergic catecholamines in vitro. Mol Pharmacol 12:409–419

    PubMed  CAS  Google Scholar 

  • Miller JA, Curella P, Zahniser NR (1988) A new densitometric procedure to measure protein levels in tissue slices used in quantitative autoradiography. Brain Res 447:60–66

    Article  PubMed  CAS  Google Scholar 

  • Molenaar P, Canale E, Summers RJ (1987) Autoradiographic localization of beta-1 and beta-2 adrenoceptors in guinea pig atrium and regions of the conducting system. J Pharmacol Exp Ther 241: 1048–1064

    PubMed  CAS  Google Scholar 

  • Molenaar P, Russell FD, Shimada T, Summers RJ (1990a) Densitometric analysis of ß1- and ß2-adrenoceptors in guinea-pig atrioventricular conducting system. J Mol Cell Cardiol 22:483–495

    Article  PubMed  CAS  Google Scholar 

  • Molenaar P, Smolich JJ, Russell FD, McMartin LR, Summers RJ (1990b) Differential regulation of beta-1 and beta-2 adrenoceptors in guinea-pig atrioventricular conducting system after chronic (−)-isoprenaline infusion. J Pharmacol Exp Ther 255:393–400

    PubMed  CAS  Google Scholar 

  • Molenaar P, Summers RJ (1987) Characterization of beta-1 and beta-2 adrenoceptors in guinea pig atrium: functional and receptor binding studies. J Pharmacol Exp Ther 241:1041–1047

    PubMed  CAS  Google Scholar 

  • Motomura S, Zerkowski H-R, Daul A, Brodde O-E (1990) On the physiologic role of beta-2 adrenoceptors in the human heart: in vitro and in vivo studies. Am Heart J 119:608–619

    Article  PubMed  CAS  Google Scholar 

  • Mukherjee C, Caron MG, Lefkowitz RJ (1976) Regulation of adenylate cyclase coupled ß-adrenergic receptors by ß-adrenergic catecholamines. Endocrinology 99:347–357

    PubMed  CAS  Google Scholar 

  • Murphree SS, Saffitz JE (1988) Delineation of the distribution of ß-adrenergic receptor subtypes in canine myocardium. Circ Res 63: 117–125

    PubMed  CAS  Google Scholar 

  • Nanoff C, Freissmuth M, Tuisl E, Schutz W (1989) A different desensitization pattern of cardiac ß-adrenoceptor subtypes by prolonged in vivo infusion of isoprenaline. J Cardiovasc Pharmacol 13:198–203

    Article  PubMed  CAS  Google Scholar 

  • Nesheim B-I (1975) Action of ß-adrenoceptor antagonists on the response to isoprenaline in the oestrogen dominated rabbit uterus. Br J Pharmacol 53:393–401

    PubMed  CAS  Google Scholar 

  • Neve KA, McGonigle P, Molinoff PB (1986) Quantitative analysis of the selectivity of radioligands for subtypes of beta adrenergic receptors. J Pharmacol Exp Ther 238:46–53

    PubMed  CAS  Google Scholar 

  • O'Donnell SR, Wanstall JC (1985) Responses to the ß2-selective agonist procaterol of vascular and atrial preparations with different functional ß-adrenoceptor populations. Br J Pharmacol 84:227–235

    PubMed  Google Scholar 

  • Ordway GA, Gambarana C, Frazer A (1988) Quantitative autoradiography of central beta adrenoceptor subtypes: comparison of the effects of chronic treatment with desipramine or centrally administered l-isoproterenol. J Pharmacol Exp Ther 247:379–389

    PubMed  CAS  Google Scholar 

  • Raposo G, Dunia I, Delavier-Klutchko C, Kaveri S, Strosberg AD, Benedetti EL (1989) Internalization of ß-adrenergic receptor in A431 cells involves non-coated vesicles. Eur J Cell Biol 50:340–352

    PubMed  CAS  Google Scholar 

  • Ruffolo RR (1982) Important concepts of receptor theory. J Acton Pharmacol 2:277–295

    Article  CAS  Google Scholar 

  • Russell FD, Molenaar P, Edyvane N, Smolich JJ, Summers RJ (1991) Autoradiographic localization and quantitation of ß-adrenoceptor subtypes in the guinea-pig sinoatrial node. Mol Neuropharmacol 1:141–147

    CAS  Google Scholar 

  • Snavely MD, Ziegler MG, Insel PA (1985) Subtype-selective down-regulation of rat renal cortical α- and ß-adrenergic receptors by catecholamines. Endocrinology 117:2182–2189

    Article  PubMed  CAS  Google Scholar 

  • Stadel JM, Strulovici B, Nambi P, Lavin TN, Briggs MM, Caron MG, Lefkowitz RJ (1983) Desensitization of the ß-adrenergic receptor of frog erythrocytes. Recovery and characterization of the down-regulated receptors in sequestered vesicles. J Biol Chem 258:3032–3038

    PubMed  CAS  Google Scholar 

  • Starke K, Montel H, Schumann HJ (1971) Influence of cocaine and phenoxybenzamine on noradrenaline uptake and release. Naunyn-Schmiedebergs Arch Pharmacol 270:210–214

    Article  PubMed  CAS  Google Scholar 

  • Stene-Larsen G, Ask JA, Helle KB, Finn R (1986) Activation of cardiac beta2 adrenoceptors in the human heart. Am J Cardiol 57:7F-10F

    Google Scholar 

  • Su YF, Harden TK, Perkins JP (1980) Catecholamine-specific desensitization of adenylate cyclase: evidence for a multistep process. J Biol Chem 255:7410–7419

    PubMed  CAS  Google Scholar 

  • Ungerer M, Bohm M, Elce JS, Erdmann E, Lohse MJ (1993) Altered expression of ß-adrenergic receptor kinase and ß1-adrenergic receptors in the failing human heart. Circulation 87:454–463

    PubMed  CAS  Google Scholar 

  • Williams DW, Summers RJ (1990) SIMUL: an accurate method for the determination of receptor subtype proportions using a personal computer. Comput Meth Programs Biomed 32:137–139

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, University of Melbourne, 3052, Parkville, Victoria, Australia

    Fraser D. Russell, Andrew R. Kompa, Peter Molenaar & Roger J. Summers

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  1. Fraser D. Russell
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  2. Andrew R. Kompa
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  3. Peter Molenaar
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  4. Roger J. Summers
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This work was supported by grants from the National Health and Medical Research Council and the National Heart Foundation of Australia

Correspondence to: R. J. Summers at the above address

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Russell, F.D., Kompa, A.R., Molenaar, P. et al. Regulation of ß-adrenoceptors in the guinea-pig sinoatrial node. Naunyn-Schmiedeberg's Arch Pharmacol 349, 463–472 (1994). https://doi.org/10.1007/BF00169134

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  • DOI: https://doi.org/10.1007/BF00169134

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