Differential regulation of rat peripheral 5-HT2A and 5-HT2B receptor systems: influence of drug treatment

M. J. Enguix1, L. Sánchez1,2, M. Villazón1,2, J. Brea1, H. Tristán1, H. J. Caruncho1, M. I. Cadavid1, M. I. Loza1
1Department of Pharmacology, School of Pharmacy, University of Santiago de Compostela, Santiago de Compostela, Spain
2Departamento de Farmacología, Facultad de Medicina, Universidad de Oviedo , Oviedo, Spain

Tóm tắt

Most studies of 5-HT2 receptor regulation have been carried out on the central nervous system (CNS) (which expresses 5-HT2A and 5-HT2C receptors); very few in vitro studies have addressed the peripheral receptors 5-HT2A and 5-HT2B. The aim of this investigation was to compare the possible short- and long-term processes regulating these peripheral receptors in the rat. The in vitro contractile response elicited by serotonin (5-HT, 10 µM) in the rat gastric fundus (5-HT2B receptor system) was rapid and followed by a partial fade to a steady state, in contrast with the rat thoracic aorta response (5-HT2A receptor system), which was more stable, slower and sustained. To characterize drug-receptor interactions, cumulative concentration/response curves (CCRCs) for 5-HT were constructed ex vivo for rat tissues treated with drugs acting at these receptors. Rats were examined 4 or 24 h after a single, i.p. administration of (±)1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane [(±)DOI, 1 or 2.5 mg/kg], clozapine, cyproheptadine or rauwolscine (10 mg/kg), 48 h after a single i.p. administration of (±)DOI (2.5 mg/kg), clozapine or cyproheptadine (10 mg/kg) or 24 h after the last of with 15 daily i.p. administrations of (±)DOI (1 or 2.5 mg/kg), clozapine, cyproheptadine or rauwolscine (10 mg/kg). In the aorta, E max (the maximum response elicited by 5-HT) was unchanged 4 h after a single dose of any of the drugs tested. However, 24 h after a single dose, E max was lower in animals treated with (±)DOI (2.5 mg/kg), clozapine or cyproheptadine than in controls, whilst 48 h after a single dose of (±)DOI (2.5 mg/kg), clozapine or cyproheptadine there was no difference in E max between experimental and control animals. After chronic treatment with (±)DOI (2.5 mg/kg), clozapine and cyproheptadine, E max was lower than in controls. In the gastric fundus, E max 4 h after a single dose of each drug was lower than in controls, and the response recovered by 24 or 48 h. Following chronic treatment, E max was significantly lower than in controls for each drug used. These findings suggest first, that regulation of peripheral 5-HT2 receptors (5-HT2A and 5-HT2B) is a functionally significant phenomenon in vivo, and occurs after administration of both agonists and antagonists. Second, the kinetics of peripheral 5-HT2 receptor regulation were similar in both in vivo and ex vivo experiments. The 5-HT2B receptors in rat gastric fundus are more sensitive to drug-induced regulation than the 5-HT2A rat aortic receptors. Finally, long-term regulation of both receptors stabilizes short-term desensitization for longer.

Tài liệu tham khảo

Akiyoshi J, Hough C, Chuang DM (1993) Paradoxical increase of 5-hydroxytryptamine2 receptors and 5-hydroxytryptamine2 receptor mRNA in cerebellar granule cells after persistent 5-hydroxytryptamine2 receptor stimulation. Mol Pharmacol 43:349–355 Akiyoshi J, Nishizono A, Yamada K, Nagayama H, Mifune K, Fujii I (1995) Rapid desensitization of serotonin 5-HT2C receptor-stimulated intracellular calcium mobilization in CHO cells transfected with cloned human 5-HT2C receptors. J Neurochem 64:2473–2479 Aloyo VJ, Dave KD, Rahman T, Harvey JA (2001) Selective and divergent regulation of cortical 5-HT(2A) receptors in rabbit. J Pharmacol Exp Ther 299:1066–1072 Anji A, Kumari M, Sullivan Hanley NR, Bryan GL, Hensler JG (2000) Regulation of 5-HT2A receptor mRNA levels and binding sites in rat frontal cortex by agonist DOI and the antagonist mianserin. Neuropharmacology 39:1996–2005 Baldessarini RJ, Centorrino F, Flood JG, Volpicelli SA, Huston-Lyons D, Cohen BM (1993) Tissue concentrations of clozapine and its metabolites in the rat. Neuropsychopharmacology 9:117–124 Baxter GS, Murphy OE, Blackburn TP (1994) Further characterization of 5-hydroxytryptamine receptors (putative 5-HT2B) in rat stomach fundus longitudinal muscle. Br J Pharmacol 112:323–331 Ben-Harari RR, Dalton BA, Osman R, Maayani S (1991) Kinetic characterization of 5-hydroxytryptamine receptor desensitization in isolated guinea-pig trachea and rabbit aorta. J Pharmacol Exp Ther 257:416–424 Berg KA, Maayani S, Goldfarb J, Scaramellini C, Leff P, Clarke WP (1998) Effector pathway-dependent relative efficacy at serotonin type 2A and 2C receptors: evidence for agonist-directed trafficking of receptor stimulus. Mol Pharmacol 54:94–104 Berg KA, Stout BD, Maayani S, Clarke WP (2001) Differences in rapid desensitization of 5-hydroxytryptamine2A and 5-hydroxytryptamine2C receptor-mediated phospholipase C activation. J Pharmacol Exp Ther 299:593–602 Blackshear MA, Sanders-Bush E (1982) Serotonin receptor sensitivity after acute and chronic treatment with mianserin. J Pharmacol Exp Ther 221:303–308 Bonhaus DW, Bach C, DeSouza A, Salazar FH, Matsuoka BD, Zuppan P, Chan HW, Eglen RM (1995) The pharmacology and distribution of human 5-hydroxytryptamine2B (5-HT2B) receptor gene products: comparison with 5-HT2A and 5-HT2C receptors. Br J Pharmacol 115:622–628 Brea J, Masaguer CF, Villazón M, Cadavid MI, Raviña E, Fontaine F, Dezi C, Pastor M, Sanz F, Loza MI (2003) Conformationally constrained butyrophenones as new pharmacological tools to study 5-HT2A and 5-HT2C receptor behaviours. Eur J Med Chem 38:433–440 Briddon SJ, Leslie RA, Elliott JM (1998) Comparative desensitization of the human 5-HT2A and 5-HT2C receptors expressed in the human neuroblastoma cell line SH-SY5Y. Br J Pharmacol 125:727–734 Buckholtz NS, Zhou DF, Freedman DX (1988) Serotonin2 agonist administration down-regulates rat brain serotonin2 receptors. Life Sci 42:2439–2445 Chen H, Li H, Chuang DM (1995) Role of second messengers in agonist up-regulation of 5-HT2A (5-HT2) receptor binding sites in cerebellar granule neurons: involvement of calcium influx and a calmodulin-dependent pathway. J Pharmacol Exp Ther 275:674–680 Clineschmidt BV, Reiss DR, Pettibone DJ, Robinson JL (1985) Characterization of 5-hydroxytryptamine receptors in rat stomach fundus. J Pharmacol Exp Ther 235:696–708 Cohen ML, Fuller RW, Wiley KS (1981) Evidence for 5-HT2 receptors mediating contraction in vascular smooth muscle. J Pharmacol Exp Ther 218:421–425 Eison AS, Eison MS, Yocca FD, Gianutsos G (1989) Effects of imipramine and serotonin-2 agonists and antagonists on serotonin-2 and beta-adrenergic receptors following noradrenergic or serotonergic denervation. Life Sci 44:1419–1427 Felder CC, Kanterman RY, Ma AL, Axelrod J (1990) Serotonin stimulates phospholipase A2 and the release of arachidonic acid in hippocampal neurons by a type 2 serotonin receptor that is independent of inositolphospholipid hydrolysis. Proc Natl Acad Sci USA 87:2187–2191 Fitzgerald LW, Burn TC, Brown BS, Patterson JP, Corjay MH, Valentine PA, Sun JH, Link JR, Abbaszade I, Hollis JM, Largent BL, Hartig PR, Hollis GF, Meunier PC, Robichaud AJ, Robertson DW (2000) Possible role of valvular serotonin 5-HT2B receptors in the cardiopathy associated with fenfluramine. Mol Pharmacol 57:75–81 Foguet M, Hoyer D, Pardo LA, Parekh A, Kluxen FW, Kalkman HO, Stuhmer W, Lubbert H (1992) Cloning and functional characterization of the rat stomach fundus serotonin receptor. EMBO J 11:3481–3487 Fukuda S, Su C, Lee TJ (1986) Mechanisms of extraneuronal serotonin uptake in the rat aorta. J Pharmacol Exp Ther 239:264–269 Furchgott RF (1972) The classification of adrenoceptors (adrenergic receptors). An evaluation from the stand point of receptor theory. In: Blaschko H, Muscholl E (eds) Catecholamines. Springer, Berlin Heidelberg New York, pp 283–335 Gray JA, Roth BL (2001) Paradoxical trafficking and regulation of 5-HT2A receptors by agonists and antagonists. Brain Res Bull 56:441–451 Gruetter CA, Lemke SM, Anestis DK, Szarek JL, Valentovic MA (1992) Potentiation of 5-hydroxytryptamine-induced contraction in rat aorta by chlorpheniramine, citalopram and fluoxetine. Eur J Pharmacol 217:109–118 Hardcastle J, Hardcastle PT, Carstairs JW, Franks CM (1994) Is desensitization of intestinal 5-hydroxytryptamine receptors an in vitro phenomenon? J Pharm Pharmacol 46:322–325 Helmeste DM, Tang SW (1983) Unusual acute effects of antidepressants and neuroleptics on S2-serotonergic receptors. Life Sci 33:2527–2533 Helton DR, Colbert WE (1994) Alterations of in-vitro 5-HT receptor pharmacology as a function of multiple treatment with 5-hydroxytryptamine or 8-hydroxy-2-(di-N-propylamino) tetralin in rat isolated aorta, uterus and fundus, and guinea-pig isolated trachea. J Pharm Pharmacol 46:902–905 Hoyer D, Clarke DE, Fozard JR, Hartig PR, Martin GR, Mylecharane EJ, Saxena PR, Humphrey PP (1994) International Union of Pharmacology classification of receptors for 5-hydroxytryptamine (serotonin). Pharmacol Rev 46:157–203 Ivins KJ, Molinoff PB (1991) Desensitization and down-regulation of 5-HT2 receptors in P11 cells. J Pharmacol Exp Ther 259:423–429 Iwaki M, Ogiso T, Fujii Y, Tanino T, Ito Y, Miki Y (1993) Pharmacokinetics of cyproheptadine and its metabolites in rats. Biol Pharm Bull 16:1276–1281 Jakab RL, Goldman-Rakic PS (1998) 5-Hydroxytryptamine2A serotonin receptors in the primate cerebral cortex: possible site of action of hallucinogenic and antipsychotic drugs in pyramidal cell apical dendrites. Proc Natl Acad Sci USA 95:735–740 Jordan BA, Devi LA (1999) G-protein-coupled receptor heterodimerization modulates receptor function. Nature 399:697–700 Kenakin T (2002) Efficacy at G-protein-coupled-receptors. Nature Rev Drug Discov 1:103–109 Kuoppamaki M, Seppala T, Syvalahti E, Hietala J (1994) Regulation of serotonin 5-HT2C receptors in the rat choroid plexus after acute clozapine treatment. Eur J Pharmacol 269:201–208 Kuoppamaki M, Palvimaki EP, Hietala J, Syvalahti E (1995) Differential regulation of rat 5-HT2A and 5-HT2C receptors after chronic treatment with clozapine, chlorpromazine and three putative atypical antipsychotic drugs. Neuropsychopharmacology 13:139–150 Kursar JD, Nelson DL, Wainscott DB, Cohen ML, Baez M (1992) Molecular cloning, functional expression, and pharmacological characterization of a novel serotonin receptor (5-hydroxytryptamine2F) from rat stomach fundus. Mol Pharmacol 42:549–557 Kursar JD, Nelson DL, Wainscott DB, Baez M (1994) Molecular cloning, functional expression, and mRNA tissue distribution of the human 5-hydroxytryptamine2B receptor. Mol Pharmacol 46:227–234 Leysen JE, Van Gompel P, Gommeren W, Woestenborghs R, Janssen PA (1986) Down regulation of serotonin-S2 receptor sites in rat brain by chronic treatment with the serotonin-S2 antagonists: ritanserin and setoperone. Psychopharmacology 88:434–44 Leysen JE, Janssen PF, Niemegeers CJ (1989) Rapid desensitization and down-regulation of 5-HT2 receptors by DOM treatment. Eur J Pharmacol 163:145–149 López-Gimenez JF, Mengod G, Palacios JM, Vilaró MT (1997) Selective visualization of rat brain 5-HT2A receptors by autoradiography with [3H]MDL 100,907. Naunyn-Schmiedeberg's Arch Pharmacol 356:446–454 López-Gimenez JF, Villazón M, Brea J, Loza MI, Palacios JM, Mengod G, Vilaro MT (2001) Multiple conformations of native and recombinant human 5-hydroxytryptamine(2a) receptors are labeled by agonists and discriminated by antagonists. Mol Pharmacol 60:690–699 Loric S, Maroteaux L, Kellermann O, Launay JM (1995) Functional serotonin-2B receptors are expressed by a teratocarcinoma-derived cell line during serotoninergic differentiation. Mol Pharmacol 47:458–466 Loza MI, Ferreiro TG, Sanz F, Lozoya E, Rodriguez J, Manaut F, Verde I, Castro E, Fontenla JA, Cadavid I, Honrubia A, Fueyo J, Raviña E (1993) Antiserotoninergic activity of 2-aminoethylbenzocyclanones in rat aorta: structure-activity relationships. J Pharm Sci 82:513–517 Manivet P, Mouillet RS, Callebert J, Nebigil CG, Maroteaux L, Hosoda S, Kellermann O, Launay JM (2000) PDZ-dependent activation of nitric-oxide syntheses by the serotonin 2B receptor. J Biol Chem 275:9324–9331 Martin GR (1998) 5-Hydroxytriptamine receptors. In: Godfraind T, Humphrey P, Ruffolo R, Vanhoutte P (eds.) The IUPHAR compendium of receptor characterization and classification. IUPHAR Media, London, pp 167–185 McKenna DJ, Nazarali AJ, Himeno A, Saavedra JM (1989) Chronic treatment with (±)DOI, a psychotomimetic 5-HT2 agonist, downregulates 5-HT2 receptors in rat brain. Neuropsychopharmacology 2:81–87 McLean PG, Coupar IM, Molenaar P (1996) Changes in sensitivity of 5-HT receptor mediated functional responses in the rat oesophagus, fundus and jejunum following chronic infusion with 5-hydroxytryptamine. Naunyn-Schmiedeberg's Arch Pharmacol 354:513–519 Mengod G, Pompeiano M, Waeber C, Sarasa M, Martinez-Mir MI, Palacios JM (1990) Studies on the differential regulation of the expression of the genes coding for 5-HT2 and 5-HT1C receptors (abstract). Eur J Pharmacol (Suppl) 183:1617 Molineaux SM, Jessell TM, Axel R, Julius D (1989) 5-HT1c receptor is a prominent serotonin receptor subtype in the central nervous system. Proc Natl Acad Sci USA 86:6793–6797 Nebigil CG, Etienne N, Schaerlinger B, Hickel P, Launay JM, Maroteaux L (2001) Developmentally regulated serotonin 5-HT2B receptors. Int J Dev Neurosci 19:365–372 Newton RA, Elliott JM (1997) Mianserin-induced down-regulation of human 5-hydroxytryptamine2A and 5-hydroxytryptamine2C receptors stably expressed in the human neuroblastoma cell line SH-SY5Y. J Neurochem 69:1031–1038 Pandey SC, Sharma RP, Janicak PG, Marks RC, Davis JM, Pandey GN (1993) Platelet serotonin-2 receptors in schizophrenia: effects of illness and neuroleptic treatment. Psychiatry Res 48:57–68 Paris G, Sugar IP, Gonsiorek W, Loza MI, Tristan H, Honrubia M, Maayani S (1997) Rapid desensitization and concurrent resensitization of the 5-HT2B receptor in rat stomach fundus. ASPET San Diego CA Pazos A, Cortes R, Palacios JM (1985) Quantitative autoradiographic mapping of serotonin receptors in the rat brain. II. Serotonin-2 receptors. Brain Res 346:231–249 Pazos A, Probst A, Palacios JM (1987) Serotonin receptors in the human brain-III. Autoradiographic mapping of serotonin-1 receptors. Neuroscience 21:97–122 Peroutka SJ, Snyder SH (1980) Long-term antidepressant treatment decreases spiroperidol-labeled serotonin receptor binding. Science 210:88–90 Pranzatelli MR (1991) Regulation of 5-HT2 receptors in rat cortex. Studies with a putative selective agonist and an antagonist. Biochem Pharmacol 42:1099–1105 Pranzatelli MR, Murthy JN, Tailor PT (1993) Novel regulation of 5-HT1C receptors: down-regulation induced both by 5-HT1C/2 receptor agonists and antagonists. Eur J Pharmacol 244:1–5 Rinaldi-Carmona M, Congy C, Simiand J, Oury DF, Soubrie P, Breliere JC, Le Fur G (1993) Repeated administration of SR 46349B, a selective 5-hydroxytryptamine2 antagonist, up-regulates 5-hydroxytryptamine2 receptors in mouse brain. Mol Pharmacol 43:84–89 Ronde P, Ansanay H, Dumuis A, Miller R, Bockaert J (1995) Homologous desensitization of 5-hydroxytryptamine4 receptors in rat esophagus: functional and second messenger studies. J Pharmacol Exp Ther 272:977–983 Roth BL, Ciaranello RD (1991) Chronic mianserin treatment decreases 5-HT2 receptor binding without altering 5-HT2 receptor mRNA levels. Eur J Pharmacol 207:169–172 Roth BL, Willins DL, Kristiansen K, Kroeze WK (1998) 5-Hydroxytryptamine2-family receptors (5-hydroxytryptamine2A, 5-hydroxytryptamine2B, 5-hydroxytryptamine2C): where structure meets function. Pharmacol Ther 79:231–257 Schmuck K, Ullmer C, Engels P, Lubbert H (1994) Cloning and functional characterization of the human 5-HT2B serotonin receptor. FEBS Lett 342:85–90 Strange PG (2002) Mechanisms of inverse agonism at G-protein-coupled receptors. Trends Pharmacol Sci 23:89–95 Thellung S, Barizza A, Maura G, Raiteri M (1993) Inhibition of the mossy fibre-granule cell glutamate transmission in rat cerebellar slices. Naunyn-Schmiedeberg's Arch Pharmacol 348:347–351 Tournois C, Mutel V, Manivet P, Launay JM, Kellermann O (1998) Cross-talk between 5-hydroxytryptamine receptors in a serotonergic cell line—involvement of arachidonic acid metabolism. J Biol Chem 273:17498–17503 Twist EC, Mitchell SN, Corn TH, Campbell IC (1991) Effect of chronic ritanserin or clorgyline on amine and metabolite levels in rat frontal cortex. Eur J Pharmacol 196:157–160 Uphouse L (1997) Multiple serotonin receptors: too many, not enough, or just the right number? Neurosci Biobehav Rev 21:679–698 Van Huizen F, Bansse MT, Stam NJ (1993) Agonist-induced down-regulation of human 5-HT1A and 5-HT2 receptors in Swiss 3T3 cells. Neuroreport 4:1327–1330 Van Oekelen D, Jurzak M, Van de Wiel D, Van Hecke G, Luyten WHML, Leysen JE (2001) Different regulation of rat 5-HT2A and rat 5-HT2C receptors in NIH 3T3 cells upon exposure to 5-HT and pipamperone. Eur J Pharmacol 425:21–32 Vane JR (1957) A sensitive method for the assay of 5-hydroxytryptamine. Br J Pharmacol 12:344–349 Villazon M, Padin JF, Cadavid MI, Enguix MJ, Tristan H, Orallo F, Loza MI (2002) Functional characterization of serotonin receptors in rat isolated aorta. Biol Pharm Bull 25:584–590 Vouret-Craviari V, Auberger P, Pouyssegur J, Van Obberghen SE (1995) Distinct mechanisms regulate 5-HT2 and thrombin receptor desensitization. J Biol Chem 270:4813–4821 Watts SW, Schenk KW, Cohen ML (1994) Selective desensitization of 5-hydroxytryptamine 2A mediated contraction in guinea pig trachea. Can J Physiol Pharmacol 72:463–470 Westphal RS, Backstrom JR, Sanders-Bush E (1995) Increased basal phosphorylation of the constitutively active serotonin 2C receptor accompanies agonist-mediated desensitization. Mol Pharmacol 48:200–205 Willins DL, Deutch AY, Roth BL (1997) Serotonin 5-HT2A receptors are expressed on pyramidal cells and interneurons in the rat cortex. Synapse 27:79–82 Willins DL, Berry SA, Alsayegh L, Backstrom JR, Sanders-Bush E, Friedman L (1999) Clozapine and other 5-hydroxytryptamine-2A receptor antagonists alter the subcellular distribution of 5-hydroxytryptamine-2A receptors in vitro and in vivo. Neuroscience 91:599–606 Xiao Z, Yao Y, Long Y, Devreotes P (1999) Desensitization of G-protein-coupled receptors. Agonist-induced phosphorylation of the chemoattractant receptor cAR1 lowers its intrinsic affinity for cAMP. J Biol Chem 274:1440–1448 Zhou XM, Fishman PH (1991) Desensitization of the human beta 1-adrenergic receptor. Involvement of the cyclic AMP-dependent but not a receptor-specific protein kinase. J Biol Chem 266:7462–7468