Abstract
The effects of 5-hydroxytryptamine (5-HT) receptor agonists and antagonists on tritium overflow evoked by high K+ were determined in superfused synaptosomes and slices, preincubated with [3H]5-HT, from guinea-pig brain cortex. In addition, we estimated the potencies of 5-HT receptor ligands in inhibiting specific [3H]5-HT binding (in the presence of 8-hydroxy-2(di-n-propylamino)tetralin and mesulergine to prevent binding to 5-HT1A and 5-HT2C sites) to guinea-pig cortical synaptosomes and membranes.
5-HT receptor agonists inhibited the K+-evoked tritium overflow from synaptosomes and slices. In synaptosomes the rank order of potencies was 2-[5-[3-(4-methylsulphonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indole-3-yl] ethylamine (L-694,247) >5-carboxamidotryptamine (5-CT) > oxymetazoline (in the presence of idazoxan) ≥ 5-HT > sumatriptan ≥ 5-methoxy-3(1,2,3,6-tetrahydropyridin-4-yl)-1H-indole (RU 24969). The potencies of the agonists in inhibiting tritium overflow from slices correlated with those in synaptosomes, suggesting that the same site of action is involved in both preparations. In synaptosomes the nonselective antagonist at cloned human 5-HT1Dα, and 5-HT1Dβ receptors, methiothepin, shifted the concentration-response curve for 5-CT to the right (apparent pA2: 7.87). In contrast, ketanserin at a concentration which should block the 5-HT1Dα, but not the 5-HT1D\, receptor did not alter the inhibitory effect of 5-CT on tritium overflow. In cortical synaptosomes and membranes, [3H]5-HT bound to a single site with high affinity. In competition experiments, 5-HT receptor agonists and antagonists inhibited specific [3H]5-HT binding. In synaptosomes the rank order was L-694,247 > methiothepin >5-CT >5-methoxytryptamine >5-HT ≥ sumatriptan ≥ oxymetazoline > RU 24969 > ketanserin > ritanserin. A very similar rank order was obtained in cerebral cortical membranes. The potencies of the 5-HT receptor agonists in inhibiting tritium overflow from synaptosomes and slices correlated with their potencies in inhibiting [3H]5-HT binding to synaptosomes and membranes.
In conclusion, the 5-HT receptors mediating inhibition of 5-HT release in the guinea-pig cortex are located on the serotoninergic axon terminals and, hence, represent presynaptic inhibitory autoreceptors. The [3H]5-HT binding sites in cerebral cortical synaptosomes and membranes exhibit the pharmacological properties of 5-HT1D receptors. The correlation between the functional responses and the binding data confirms the 5-HT1D character of the presynaptic 5-HT autoreceptors. According to the results of the interaction experiment of ketanserin and methiothepin with 5-CT on 5-HT release, the presynaptic 5-HT autoreceptors can be subclassified as 5-HT1D\-like.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beer MS, Stanton JA, Bevan Y, Chauhan NS, Middlemiss DN (1992) An investigation of the 5-HT1D receptor binding affinity of 5-hydroxytryptamine, 5-carboxamidotryptamine and sumatriptan in the central nervous system of seven species. Fur J Pharmacol 213: 193–197
Beer MS, Stanton JA, Bevan Y, Heald A, Reeve AJ, Street LJ, Matassa VG, Hargreaves RJ, Middlemiss DN (1993) L-694,247: a potent 5-HT1D receptor agonist. Br J Pharmacol 110:1196–1200
Boess FG, Martin IL (1994) Molecular biology of 5-HT receptors. Neuropharmacology 33: 275–317
Briley M, Moret C (1993) 5-HT and antidepressants: in vitro and in vivo release studies. Trends Pharmacol Sci 14:396–397
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
Bühlen M, Fink K, Göthert M (1995) Presynaptic location of inhibitory 5-hydroxytryptamine (5-HT) autoreceptors in the guinea-pig brain cortex: an investigation of [3H]5-HT release and binding in synaptosomes. Naunyn-Schmiedeberg's Arch Pharmacol 351:R142
Cerrito F, Raiteri M (1979) Scrotonin release is modulated by presynaptic autoreceptors. Eur J Pharmacol 57:427–430
Del Arco C, Galende I, Pazos A (1993) Autoradiographic mapping of 5-HT1 receptors in the guinea-pig brain with particular reference to the 5-HT1D receptor sites. Naunyn-Schmiedeberg's Arch Pharmacol 347:248–256
Fink K, Zentner J, Göthert M (1995) Subclassification of presynaptic 5-HT autoreceptors in the human cerebral cortex as 5-HT1D\ receptors. Naunyn-Schmiedeberg's Arch Pharmacol 352:451–454
Furchgott RF (1972) The classification of adrenoceptors (adrenergic receptors). An evaluation from the standpoint of receptor theory. In: Blaschko H, Muscholl E (eds) Handbook of experimental pharmacology, vol XXXIII. Springer, Berlin Heidelberg New York, pp 283–335
Galzin AM, Poirier MF, Lista A, Chodkiewicz JP, Blier P, Ramadine R, Loo 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
Göthert M, Schlicker E (1993) Relevance of 5-HT autoreceptors for psychotropic drug action. In: Gram LF, Balant LP, Meltzer HY, Dahl SG (eds) Clinical pharmacology in psychiatry. Springer, Heidelberg New York London, pp 38–51
Göthert M, Schlicker E, Fink K, Classen K (1987) Effects of RU 24969 on serotonin release in rat brain cortex: further support for the identity of serotonin autoreceptors with 5-HT1B sites. Arch Int Pharmacodyn Ther 288:31–42
Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electron microscope study of cell fragments derived by homogenization and centrifugation. J Anat (Lond) 96:79–88
Hartig PR, Branchek TA, Weinshank RL (1992) A subfamily of 5-HTID receptor genes. Trends Pharmacol Sci 13:152–159
Hoyer D, Middlemiss DN (1989) Species differences in the pharmacology of terminal 5-HT autoreceptors in mammalian brain. Trends Pharmacol Sci 10:130–132
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
Leonhardt S, Herrik-Davis K, Titeler M (1989) Detection of a novel serotonin receptor subtype (5-HT1E) in human brain: interaction with a GTP-binding protein. J Neurochem 53:465–471
Limberger N, Deicher R, Starke K (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
Lowry OH, Rosebrough NJ, Farr AK, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275
Mahle CD, Nowak HP, Mattson RJ, Hurt SD, Yocca FD (1991) [3H]5-Carboxamidotryptamine labels multiple high affinity 5-HT1D-like sites in guinea pig brain. Eur J Pharmacol 205:323–324
Matthes H, Boschert U, Amlaiky N, Grailhe R, Plassat JL, Muscatelli F, Mattei MG, Hen R (1993) Mouse 5-hydroxytryptamine5A and 5-hydroxytryptamine5B receptors define a new family of serotonin receptors: cloning, functional expression, and chromosomal localization. Mol Pharmacol 43:313–319
Maura G, Thellung S, Andrioli GC, Ruelle A, Raiteri I (1993) Release-regulating serotonin 5-HT1D autoreceptors in human cerebral cortex. J Neurochem 60:1179–1182
McAllister G, Charlesworth A, Snodin C, Beer MS, Noble AJ, Middlemiss DN, Iversen LL, Whiting P (1992) Molecular cloning of a serotonin receptor from human brain (5-HTIE): A fifth 5-HT1-like subtype. Proc Natl Acad Sci USA 89:5517–5521
Middlemiss DN (1988) Autoreceptors regulating serotonin release. In: Sanders-Bush E (ed) The serotonin receptors. Humana Press, Clifton, NJ, pp 210–224
Middlemiss DN, Bremer ME, Smith SM (1988)A pharmacological analysis of the 5-HT receptor mediating inhibition of 5-HT release in the guinea-pig frontal cortex. Eur J Pharmacol 157:101–107
Molderings GJ, Frölich D, Likungu J, Göthert M (1996) Inhibition of noradrenaline release via presynaptic 5-HT1Dα, receptors in human atrium. Naunyn Schmiedeberg's Arch Pharmacol 353:272–280
Moret C (1985) Pharmacology of the serotonin autoreceptor. In: Green AR (ed) Neuropharmacology of serotonin. Oxford University Press, Oxford, pp 21–49
Nowak HP, Mahle CD, Yocca FD (1993) [3H]-5-carboxamidotryptamine labels 5-HT1D binding sites in bovine substantia nigra. Br J Pharmacol 109:1206–1211
Pauwels PJ, Reihsaus E, Palmier C, Journot L, Lyons J, Colpaert FC (1995) Ketanserin differentiates between cloned human 5-HT1Dα receptors and cloned human 5-HT1D\,, sheep and bovine caudate nucleus 5-HT1D receptor sites. Cell Pharmacol 2:1–9
Schipper J, Tulp MTM (1988) Serotonin autoreceptors in guinea pig cortex slices resemble the 5-HT1d binding site. Soc Neurosci Abstr 14:552
Schoeffter P, Hoyer D (1991) Interaction of the α-adrenoceptor agonist oxymetazoline with serotonin 5-HT1A, 5-HT1B, 5-HT1C and 5-HT1D receptors. Eur J Pharmacol 196:213–216
Starke K, Göthert M, Kilbinger H (1989) Modulation of neurotransmitter release by presynaptic autoreceptors. Physiol Rev 69:864–989
Sumner MJ, Humphrey PPA (1989) 5-HT1D binding sites in porcine brain can be sub-divided by GR43175. Br J Pharmacol 98:29–31
To ZP, Bonhaus DW, Eglen RM, Jakeman LB (1995) Characterization and distribution of putative 5-ht7 receptors in guinea-pig brain. Br J Pharmacol 115:107–116
Waeber C, Schoeffter P, Palacios JM, Hoyer D (1988) Molecular pharmacology of 5-HT1D recognition sites: radioligand binding studies in human, pig and calf brain membranes. Naunyn-Schmiedeberg's Arch Pharmacol 337:595–601
Waeber C, Schoeffter P, Palacios JM, Hoyer D (1989) 5-HT1D receptors in guinea-pig and pigeon brain. Naunyn-Schmiedeberg's Arch Pharmacol 340:479–485
We inshank RL, Branchek T, Hartig PR (1991) DNA encoding human 5-HT1D receptors and uses thereof. International Patent No. WO 91/17174
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bühlen, M., Fink, K., Böing, C. et al. Evidence for presynaptic location of inhibitory 5-HT1D\-like autoreceptors in the guinea-pig brain cortex. Naunyn-Schmiedeberg's Arch Pharmacol 353, 281–289 (1996). https://doi.org/10.1007/BF00168629
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00168629