Abstract
A clonal cell line derived from rat renal mesangial cells was shown to express endogenous 5-hydroxytryptamine (serotonin, 5-HT) receptors that mediate inhibition of cyclic AMP accumulation. These receptors were characterized as being of the 5-HT1B receptor subtype. 5-HT1 receptor agonists inhibited forskolin-stimulated cyclic AMP accumulation in rat renal mesangial cells (60–70% maximal inhibition) with the following rank order of potency (mean pEC50 values±SEM, n ≥ 3): ergotamine (9.58±0.51)>RU 24969 (8.67±0.23)≥5-CT (8.42±0.06)≥CP 93129 (8.15±0.27)>5-HT (7.75±0.11) > sumatriptan (6.29±0.30) > 8-OH-DPAT (4.32±0.15). 5-HT2 and 5-HT4 receptor agonists were without effect. 5-HT-induced inhibition of cyclic AMP accumulation was abolished by a pre-treatment of the cells with pertussis toxin. (-)Propranolol was a partial agonist (27% maximal inhibition, pEC50 7.19±0.24, n = 3); when used as an antagonist at 1 μM, it shifted the concentration-response curve of 5-HT to the right (pKB 7.22±0.35, n = 3). Methiothepin was a competitive antagonist of 5-HT (pA2 8.04±0.10, Schild slope 0.87±0.21, n = 3). Rauwolscine (10 μM) had no antagonist activity. There was a significant correlation (r = 0.98, P = 0.0001) between the cyclic AMP data obtained in rat mesangial cells and 5-HT1B binding data reported in rat brain cortex. The same pattern of responses was observed in early passages of primary cultures of rat mesangial cells. This study shows that rat mesangial cells can be used as a convenient source of functional 5-HT1B receptors. It also constitutes further evidence for the widespread distribution of 5-HT1B receptors outside the brain.
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References
Adham N, Ellerbrook B, Hartig P, Weinshank RL, Branchek T (1993) Receptor reserve masks partial agonist activity of drugs in a cloned rat 5-hydroxytryptamine1B receptor expression system. Mol Pharmacol 43:427–433
Arunlakshana O, Schild HO (1959) Some quantitative uses of drug antagonists. Br J Pharmacol Chemother 14:48–58
Bouhelal R, Smounya R, Bockaert J (1988) 5-HT1B receptors are negatively coupled with adenylate cyclase in rat substantia nigra. Eur J Pharmacol 151:189–196
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
Craig DA, Martin GR (1993) 5-HT1B receptors mediate potent contractile responses to 5-HT in rat caudal artery. Br J Pharmacol 109:609–611
DeLean 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
Del Greco F, Masson GMC, Corcoran AC (1956) Renal and arterial effects of serotonin in the anesthetized rat. Am J Physiol 187:509–514
Dumuis A, Sebben M, Bockaert J (1989) BRL 24929: a potent agonist at a non-classical 5-HT receptor positively coupled with adenylate cyclase in colliculi neurons. Eur J Pharmacol 162:381–384
Engel G, Göthert M, Hoyer D, Schlicker E, Hillenbrand K (1986) Identity of inhibitory presynaptic 5-hydroxytryptamine (5-HT) autoreceptors in the rat brain cortex with 5-HT1B binding sites. Naunyn-Schmiedeberg's Arch Pharmacol 332:1–7
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
Hartig P, Branchek TA, Weinshank RL (1992) A subfamily of 5-HT1D receptor genes. Trends Pharmacol Sci 13:152–159
Heuring RE, Schlegel JR, Peroutka SJ (1986) Species variations in RU 24969 interactions with non-5-HT1A binding sites. Eur J Pharmacol 122:279–282
Hoyer D, Engel G, Kalkman HO (1985) Characterization of the 5-HT1B recognition site in rat brain: binding studies with (-)[125I]iodocyanopindolol. Eur J Pharmacol 118:1–12
Hoyer D, Schoeffter P, Gray JA (1989a) A comparison of the interactions of dihydroergotamine, ergotamine and GR 43175 with 5-HT1 receptor subtypes. Cephalalgia 9 [Suppl 10]:340–341
Hoyer D, Waeber C, Schoeffter P, Palacios JM, Dravid A (1989b) 5-HT1C receptor-mediated stimulation of inositol phosphate production in pig choroid plexus. Naunyn-Schmiedeberg's Arch Pharmacol 339:252–258
Humphrey PPA, Hartig P, Hoyer D (1993) A proposed new nomenclature for 5-HT receptors. Trends Pharmacol Sci 14:233–236
Jazayeri A, Meyer WJ III, Kent TA (1989) 5-HT1B and 5-HT2 serotonin binding sites in cultured Wistar-Kyoto rat aortic smooth muscle cells. Eur J Pharmacol 169:183–187
Macor JE, Burkhart CA, Heym JE, Ives JL, Lebel LA, Newman ME, Nielsen JA, Ryan K, Schulz DW, Torgersen LK, Koe BK (1990) 3-(1,2,5,6-Tetrahydropyrid-4-yl)pyrrolo[3,2-b]pyrid-5-one: a potent and selective serotonin (5-HT1B) agonist and rotationally restricted phenolic analogue of 5-methoxy-3-(1,2,5,6-tetrahydropyrid-4-yl)indole. J Med Chem 33:2087–2093
Maura G, Ulivi M, Raiteri M (1987) (-)Propranolol and (±)cyano-pindolol are mixed agonists-antagonists at serotonin autoreceptors in the hippocampus of the brain. Neuropharmacology 26:713–717
Maura G, Raiteri M (1986) Cholinergic terminals in rat hippocampus possess 5-HT1B receptors mediating inhibition of acetylcholine release. Eur J Pharmacol 129:333–337
Middlemiss DN, Fozard JR (1983) 8-Hydroxy-2-(di-n-propylamino)-tetralin discriminates between subtypes of the 5-HT1 recognition site. Eur J Pharmacol 90:151–153
Molderings GJ, Fink K, Schlicker E, Göthert M (1987) Inhibition of noradrenaline release via presynaptic 5-HT1B receptors of the rat vena cava. Naunyn-Schmiedeberg's Arch Pharmacol 336:245–250
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
Pazos A, Palacios JM (1985) Quantitative autoradiographic mapping of serotonin receptors in the rat brain. I. Serotonin-1 receptors. Brain Res 346:205–230
Pfeilschifter J (1990) Angiotensin II B-type receptor mediates phosphoinositide hydrolysis in mesangial cells. Eur J Pharmacol 184:201–202
Pfeilschifter J, Kurtz A, Bauer C (1986) Role of phospholipase C and protein kinase C in vasoconstrictor-induced prostaglandin synthesis in cultured rat renal mesangial cells. Biochem J 234:125–130
Pfeilschifter J, Merriweather C (1993) Extracellular ATP and UTP activation of phospholipase D is mediated by protein kinase C-ε in rat renal mesangial cells. Br J Pharmacol 110:847–853
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
Schoeffter P, Hoyer D (1989a) 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
Schoeffter P, Hoyer D (1989b) 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
Schoeffter P, Pfeilschifter J, Bobirnac I (1994) 5-Hydroxytryptamine 5-HT1B receptors mediate inhibition of adenylyl cyclase in a rat renal mesangial cell line. Br J Pharmacol 112:110P
Seuwen K, Magnaldo I, Pouysségur J (1988) Serotonin stimulates DNA synthesis in fibroblasts acting through 5-HT1B receptors coupled to a G1-protein. Nature 335:254–256
Takuwa N, Ganz M, Takuwa Y, Sterzel RB, Rasmussen H (1989) Studies of the mitogenic effect of serotonin in rat renal mesangial cells. Am J Physiol 257:F431-F439
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Schoeffter, P., Pfeilschifter, J. & Bobirnac, I. 5-Hydroxytryptamine 5-HT1B receptors inhibiting cyclic AMP accumulation in rat renal mesangial cells. Naunyn-Schmiedeberg's Arch Pharmacol 351, 35–39 (1995). https://doi.org/10.1007/BF00169061
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DOI: https://doi.org/10.1007/BF00169061