Skip to main content
SpringerLink
Log in

5-Hydroxytryptamine 5-HT1D receptors mediating inhibition of cyclic AMP accumulation in Madin-Darby canine kidney (MDCK) cells

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

5-Hydroxytryptamine 5-HT1B/5-HT1D receptors are members of the same receptor subfamily, but display a different pharmacology (Hartig et al. (1992) Trends Pharmacol Set 13:152–159). Whereas several cell lines have been reported to contain 5-HT1B receptors, none has been described, however, that endogenously expresses well-characterized 5-HT1D receptors. The present study deals with the identification of 5-HT1D receptors inhibiting cyclic AMP accumulation in Madin-Darby canine kidney (MDCK) cells. 5-HT (1 nM− 10 μM) induced a concentration-dependent inhibition of the cyclic AMP accumulation stimulated by prostaglandin E1 (1 μM) in MDCK cells. The maximal effect of 5-HT averaged 50% inhibition and was abolished after a pre-treatment of the cells with pertussis toxin. Other agonists mimicked the effects of 5-HT, with the following rank order of potency (pEC50 ± SEM, n ≥ 3): 5-carboxamidotryptamine (8.36 ± 0.48) > PAPP (p-aminophenylethyl-m-trifluoromethylphenyl piperazine, 7.89 ± 0.23) > 5-HT (7.35 ± 0.05) > sumatriptan (6.65 ± 0.27). PAPP behaved as a partial agonist. 8-OH-DPAT (8-hydroxy-2(di-n-propylamino)tetralin) was less potent, its maximal effect being not reached at 0.1 mM. Methiothepin, GR127935, (−)propranolol, rauwolscine and ketanserin were all devoid of intrinsic activity (up to 10 μM or 0.1 mM). Methiothepin (10 nM, 0.1 μM and 1 μM) antagonized 5-HT effect (pA2 8.57 ± 0.44, Schild slope 1.17 ± 0.21, n = 3). GR127935 (1 nM, 10 nM and 0.1 μM) shifted the curve of 5-HT to the right, but the antagonism was not fully surmountable (apparent pKB value, 9.80 ± 0.16, n = 9). From the shifts obtained with rauwolscine (1 μM) and (−)propranolol (10 μM), respective pKB values were estimated 6.68 ± 0.30 and ≈ 5.4 (n = 3 each). PAPP, when tested as an antagonist at 1 μM, also shifted the curve of 5-HT to the right, with a pKB of 8.27 ± 0.16 (n = 3). Finally, ketanserin (10 μM) also antagonized the effects of 5-HT, the pKB being 6.54 ± 0.16 (n = 9). The rank orders of agonist and antagonist potencies strongly suggest 5-HT receptors mediating inhibition of cyclic AMP accumulation in MDCK cells to be 5-HT1D receptors. This is the first report of a cell line expressing endogenous, well-characterized, 5-HT1D receptors. With regard to the 5-HT1D receptor subtype involved, the relatively high potency of ketanserin would suggest it to be a 5-HT1Dα subtype or a mixture of 5-HT1Dα/5-HT1D\ subtypes. However, caution must be exercised here, owing to the poor knowledge of canine 5-HT1D receptor subtypes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Arunlakshana O, Schild HO (1959) Some quantitative uses of drug antagonists. Br J Pharmacol Chemother 14:48–58

    Google Scholar 

  • Bockaert J, Dumuis A, Bouhelal R, Sebben M, Cory RN (1987) Piperazine derivatives including the putative anxiolytic drugs, buspirone and ipsapirone, are agonists at 5-HT1A receptors negatively coupled with adenylate cyclase in hippocampal neurones. Naunyn-Schmiedeberg's Arch Pharmacol 335:588–592

    Google Scholar 

  • Bradley PB, Engel G, Feniuk W, Fozard JR, Humphrey PPA, Middlemiss DN, Mylecharane EJ, Richardson BP, Saxena PR (1986) Proposals for the classification and nomenclature of functional receptors for 5-hydroxytryptamine. Neuropharmacology 25:563–576

    Google Scholar 

  • Bruinvels AT, Lery H, Nozulak J, Palacios JM, Hoyer D (1992) 5-HT1D binding sites in various species: similar pharmacological profile in dog, monkey, calf, guinea-pig and human brain membranes. Naunyn-Schmiedeberg's Arch Pharmacol 346:243–248

    Google Scholar 

  • Cattaneo MG, Palazzi E, Bondiolotti G, Vicentini LM (1994) 5HT1D receptor type is involved in stimulation of cell proliferation by serotonin in human small cell lung carcinoma. Eur J Pharmacol Mol Pharmacol Sect 268:425–430

    Google Scholar 

  • Ciaranello RD, Tan GL, Dean R (1990) G-protein-linked serotonin receptors in mouse kidney exhibit identical properties to 5-HT1B receptors in brain. J Pharmacol Exp Ther 252:1347–1354

    Google Scholar 

  • Clitherow JW, Scopes DIC, Skingle M, Jordan CC, Feniuk W, Campbell IB, Carter MC, Collington EW, Connor HE, Higgins GA, Beattie D, Kelly HA, Mitchell WL, Oxford AW, Wadsworth AH, Tyers MB (1994) Evolution of a novel series of [(N,N-dimethylamino)propyl]-and piperazinylbenzanilides as the first selective 5-HT1D antagonists. J Med Chem 37:2253–2257

    Google Scholar 

  • DeLéan A, Stadel JM, Lefkowitz RJ (1980) A ternary complex model explains the agonist-specific binding properties of the adenylate cyclase-coupled \-adrenergic receptor. J Biol Chem 255: 7108–7117

    Google Scholar 

  • De Vivo M, Maayani S (1986) Characterization of the 5-hydroxytryptamine1A receptor-mediated inhibition of forskolin-stimulated adenylate cyclase activity in guinea-pig and rat hippocampal membranes. J Pharmacol Exp Ther 238:248–253

    Google Scholar 

  • Ebersole BJ, Diglio CA, Kaufman DW, Berg KA (1993) 5-Hydroxytryptamine1-like receptors linked to increases in intracellular calcium concentration and inhibition of cyclic AMP accumulation in cultured vascular smooth muscle cells derived from bovine basilar artery. J Pharmacol Exp Ther 266:692–699

    Google Scholar 

  • Gausch CR, Hard WL, Smith TF (1966) Characterization of an established line of canine kidney cells (MDCK). Proc Soc Exp Biol Med 122:931–935

    Google Scholar 

  • Galzin AM, Poirier MF, Lista A, Chodkiewicz JP, Blier P, Ramdine R, Loô H, Roux FX, Redondo A, Langer SZ (1992) Characterization of the 5-hydroxytryptamine receptor modulating the release of 5-[3H]hydroxytryptamine in slices of the human neocortex. J Neurochem 59:1293–1301

    Google Scholar 

  • Giles H, Lansdell SJ, Fox P, Lockyer M, Hall V, Martin GR (1994) Characterisation of a 5-HT1B receptor on CHO cells: functional responses in the absence of radioligand binding. Br J Pharmacol 112:317P

  • Hamblin MW, Metcalf MA (1991) Primary structure and functional characterization of a human 5-HT1D-type serotonin receptor. Mol Pharmacol 40:143–148

    Google Scholar 

  • Hamel E, Bouchard D (1991) Contractile 5-HT1 receptors in human isolated pial arterioles: correlation with 5-HT1D binding sites. Br J Pharmacol 102:227–233

    Google Scholar 

  • Hamel E, Fan E, Linville D, Ting V, Villemure J-G, Chia L-S (1993) Expression of mRNA for the serotonin 5-hydroxytryptamine1D\receptor subtype in human and bovine cerebral arteries. Mol Pharmacol 44:242–246

    Google Scholar 

  • Harel-Dupas C, Cloëz I, Fillion G (1991) The inhibitory effect of trifluoromethylphenylpiperazine on [3H]acetylcholine release in guinea-pig hippocampal synaptosomes is mediated by a 5-hydroxytryptamine1 receptor distinct from 1A, 1B, and 1C subtypes. J Neurochem 56:221–227

    Google Scholar 

  • Hartig P, Branchek TA, Weinshank RL (1992) A subfamily of 5-HT1D receptor genes. Trends Pharmacol Sci 13:152–159

    Google Scholar 

  • Herzlinger DA, Easton TG, Ojakian GK (1982) The MDCK epithelial line expresses a cell surface antigen of the kidney distal tubule. J Cell Biol 93:269–277

    Google Scholar 

  • Heuring RE, Peroutka SJ (1987) Characterization of a novel 3H-5-hydroxytryptamine binding site subtype in bovine brain membranes. J Neurosci 7:894–903

    Google Scholar 

  • Hoyer D (1988) 5-Hydroxytryptamine receptors and effector coupling mechanisms in peripheral tissues. In: Fozard JR (ed) The peripheral actions of 5-hydroxytryptamine. Oxford University Press, Oxford New York Tokyo, pp 72–99

    Google Scholar 

  • Hoyer D, Middlemiss DN (1989) Species differences in the pharmacology of terminal 5-HT autoreceptors in mammalian brain. Trends Pharmacol Sci 10:130–132

    Google Scholar 

  • Hoyer D, Clarke DE, Fozard JR, Hartig PR, Martin GR, Mylecharane EJ, Saxena PR, Humphrey PPA (1994) International Union of Pharmacology classification of receptors for 5-hydroxytryptamine (serotonin). Pharmacol Rev 46:157–203

    Google Scholar 

  • Humphrey PPA, Hartig P, Hoyer D (1993) A proposed new nomenclature for 5-HT receptors. Trends Pharmacol Sci 14:233–236

    Google Scholar 

  • Kaumann AJ, Frenken M, Posival H, Brown AM (1994) Variable participation of 5-HT1-like receptors and 5-HT2 receptors in serotonin-induced contraction of human isolated coronary arteries. 5-HT1-like receptors resemble cloned 5-HT1D\ receptors. Circulation 90:1141–1153

    Google Scholar 

  • Libert F, Parmentier M, Lefort A, Dinsart C, Van Sande J, Maenhaut C, Simons MJ, Dumont JE, Vassart G (1989) Selective amplification and cloning of four new members of the G-protein-coupled receptor family. Science 244:569–572

    Google Scholar 

  • Limberger N, Deicher R, Starke (1991) Species differences in presynaptic serotonin autoreceptors: mainly 5-HT1B but possibly in addition 5-HT1D in the rat, 5-HT1D in the rabbit and guinea-pig brain cortex. Naunyn-Schmiedeberg's Arch Pharmacol 343:353–364

    Google Scholar 

  • Maenhaut C, Van Sande J, Massart C, Dinsart C, Libert F, Monferini E, Giraldo E, Ladinsky H, Vassart G, Dumont JE (1991) The orphan receptor cDNA RDC4 encodes a 5-HT1D serotonin receptor. Biochem Biophys Res Commun 180:1460–1468

    Google Scholar 

  • Mangoo-Karim R, Uchic ME, Grant M, Shumate WA, Calvet JP, Park CH, Grantham JJ (1989) Renal epithelial fluid secretion and cyst growth: the role of cyclic AMP. FASEB J 3:2629–2632

    Google Scholar 

  • Middlemiss DN, Bremer ME, Smith SM (1988) A pharmacological analysis of the 5-HT receptors mediating inhibition of 5-HT release in the guinea-pig frontal cortex. Eur J Pharmacol 157:101–107

    Google Scholar 

  • Molderings GJ, Werner K, Likungu J, Göthert M (1990) Inhibition of noradrenaline release from the sympathetic nerves of the human saphenous vein via presynaptic 5-HT receptors similar to the 5-HT1D subtype. Naunyn-Schmiedeberg's Arch Pharmacol 342:371–377

    Google Scholar 

  • Murphy TJ, Bylund DB (1989) Characterization of serotonin-1B receptors negatively coupled to adenylate cyclase in OK cells, a renal epithelial cell line from the Opossum. J Pharmacol Exp Ther 249:535–543

    Google Scholar 

  • Salomon Y (1991) Cellular responsiveness to hormones and neurotransmitters: conversion of [3H]adenine to [3H]cAMP in cell monolayers, cell suspension, and tissue slices. Methods Enzymol 195:22–28

    Google Scholar 

  • Schlicker E, Fink K, Göthert M, Hoyer D, Molderings G, Roschke I, Schoeffter P (1989) The pharmacological properties of the presynaptic serotonin autoreceptorin the pig brain cortex conform to the 5-HT1D receptor subtype. Naunyn-Schmiedeberg's Arch Pharmacol 340:45–51

    Google Scholar 

  • Schoeffter P, Hoyer D (1989a) Interaction of arylpiperazines with 5-HT1A, 5-HT1B, 5-HT1C and 5-HT1D receptors: do discriminatory 5-HT1B receptor ligands exist? Naunyn-Schmiedeberg's Arch Pharmacol 339:675–683

    Google Scholar 

  • Schoeffter P, Hoyer D (1989b) How selective is GR 43175? Interactions with functional 5-HT1A, 5-HT1B, 5-HT1C and 5-HT1D receptors. Naunyn-Schmiedeberg's Arch Pharmacol 340:135–138

    Google Scholar 

  • Schoeffter P, Hoyer D (1989c) 5-Hydroxytryptamine 5-HT1B and 5-HT1D receptors mediating inhibition of adenylate cyclase activity. Pharmacological comparison with special reference to the effects of yohimbine, rauwolscine and some \-adrenoceptor antagonists. Naunyn-Schmiedeberg's Arch Pharmacol 340: 285–292

    Google Scholar 

  • Schoeffter P, Waeber C, Palacios JM, Hoyer D (1988) The 5-hydroxytryptamine 5-HT1D receptor subtype is negatively coupled to adenylate cyclase in calf substantia nigra. Naunyn-Schmiedeberg's Arch Pharmacol 337:602–608

    Google Scholar 

  • Schoeffter P, Pfeilschifter J, Bobirnac I(1995) 5-Hydroxytryptamine 5-HT1B receptors inhibiting cyclic AMP accumulation in rat renal mesangial cells. Naunyn-Schmiedeberg's Arch Pharmacol 351:35–39

    Google Scholar 

  • Seuwen K, Magnaldo I, Pouysségur J (1988) Serotonin stimulates DNA synthesis in fibroblasts acting through 5-HT1B receptors coupled to a Gi-protein. Nature 335:254–256

    Google Scholar 

  • Skingle M, Scopes DIC, Feniuk W, Connor HE, Carter MC, Clitherow JW, Tyers MB (1993) GR127935: a potent orally active 5-HT1D receptor antagonist. Br J Pharmacol 110:9P

  • Waeber C, Schoeffter P, Palacios JM, Hoyer D (1988) Molecular pharmacology of 5-HTP1D recognition sites: radioligand binding studies in human, pig and calf brain membranes. Naunyn-Schmiedeberg's Arch Pharmacol 337:595–601

    Google Scholar 

  • Waeber C, Schoeffter P, Palacios JM, Hoyer D (1989) 5-HT1D receptors in guinea-pig and pigeon brain. Radioligand binding and biochemical studies. Naunyn-Schmiedeberg's Arch Pharmacol 340:479–485

    Google Scholar 

  • Weinshank RL, Zgombick JM, Macchi MJ, Branchek TA, Hartig PR (1992) Human serotonin 1D receptor is encoded by a subfamily of two distinct genes: 5-HT1Dα and 5-HT1D\. Proc Natl Acad Sci USA 89:3630–3634

    Google Scholar 

  • Zgombick JM, Weinshank RL, Macchi M, Schechter LE, Branchek TA, Hartig PR (1991) Expression and pharmacological characterization of a canine 5-hydroxytryptamine1D receptor subtype. Mol Pharmacol 40:1036–1042

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Preclinical Research 386/744, Sandoz Pharma Ltd., CH-4002, Basel, Switzerland

    P. Schoeffter & I. Bobirnac

Authors
  1. P. Schoeffter
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I. Bobirnac
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schoeffter, P., Bobirnac, I. 5-Hydroxytryptamine 5-HT1D receptors mediating inhibition of cyclic AMP accumulation in Madin-Darby canine kidney (MDCK) cells. Naunyn-Schmiedeberg's Arch Pharmacol 352, 256–262 (1995). https://doi.org/10.1007/BF00168555

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00168555

Key words

Search

Navigation