Characterisation of 5-HT3 Recognition Sites in Membranes of NG 108-15 Neuroblastoma-glioma Cells with [3H]ICS 205-930

Overview

Journal Naunyn Schmiedebergs Arch Pharmacol

Specialty Pharmacology

Date 1988 May 1

PMID 3412489

Citations 9

Authors

H C Neijt

A Karpf

P Schoeffter

G Engel

D Hoyer

Affiliations

Soon will be listed here.

Abstract

1. The binding characteristics of [3H]ICS 205-930, a potent and selective 5-hydroxytryptamine 5-HT3 receptor antagonist, were investigated in membranes prepared from murine neuroblastoma-glioma NG 108-15 cells. 2. [3H]ICS 205-930 bound rapidly, reversibly and stereoselectively to a homogeneous population of high affinity recognition sites: Bmax = 58 +/- 3 fmol/mg protein, pKD = 9.01 +/- 0.08 (n = 11). Non linear regression and Scatchard analysis of saturation data suggested the existence of a single class of [3H]ICS 205-930 recognition sites on NG 108-15 cells. The binding was rapid, stable and reversible. The affinity of [3H]ICS 205-930 determined in kinetic studies was in agreement with that obtained under equilibrium conditions. 3. Competition studies performed with a variety of agonists and antagonists also suggested the presence of a homogeneous population of [3H]ICS 205-930 recognition sites. All competition curves were steep and monophasic and were best fit by a 1 receptor site model. [3H]ICS 205-930 binding sites displayed the pharmacological profile of a 5-HT3 receptor. Potent 5-HT3 receptor antagonists showed nanomolar affinities for [3H]ICS 205-930 binding sites with the following rank order of potency: SDZ 206-830 greater than ICS 205-930 greater than SDZ 206-792 greater than BRL 43694 greater than quipazine greater than BRL 24924 greater than SDZ 210-204 greater than MDL 72222 greater than SDZ 210-205. Metoclopramide, mCP and mianserin showed submicromolar affinity.(ABSTRACT TRUNCATED AT 250 WORDS)

Citing Articles

The binding orientations of structurally-related ligands can differ; A cautionary note.

Ruepp M, Wei H, Leuenberger M, Lochner M, Thompson A Neuropharmacology. 2017; 119:48-61.

PMID: 28137449 PMC: 5464333. DOI: 10.1016/j.neuropharm.2017.01.023.

The interaction of trichloroethanol with murine recombinant 5-HT3 receptors.

Downie D, Hope A, Belelli D, Lambert J, Peters J, Bentley K Br J Pharmacol. 1995; 114(8):1641-51.

PMID: 7541281 PMC: 1510386. DOI: 10.1111/j.1476-5381.1995.tb14952.x.

[3H]ICS 205-930 labels 5-HT3 recognition sites in membranes of cat and rabbit vagus nerve and superior cervical ganglion.

Hoyer D, Waeber C, Karpf A, Neijt H, Palacios J Naunyn Schmiedebergs Arch Pharmacol. 1989; 340(4):396-402.

PMID: 2586633 DOI: 10.1007/BF00167040.

The gastrointestinal prokinetic benzamide derivatives are agonists at the non-classical 5-HT receptor (5-HT4) positively coupled to adenylate cyclase in neurons.

Dumuis A, Sebben M, Bockaert J Naunyn Schmiedebergs Arch Pharmacol. 1989; 340(4):403-10.

PMID: 2555720 DOI: 10.1007/BF00167041.

Molecular size of the 5-HT3 receptor solubilized from NCB 20 cells.

McKernan R, Biggs C, Gillard N, Quirk K, Ragan C Biochem J. 1990; 269(3):623-8.

PMID: 2390056 PMC: 1131632. DOI: 10.1042/bj2690623.

References

Hoyer D, Neijt H . Identification of serotonin 5-HT3 recognition sites by radioligand binding in NG108-15 neuroblastoma-glioma cells. Eur J Pharmacol. 1987; 143(2):291-2. DOI: 10.1016/0014-2999(87)90547-4. View

Round A, WALLIS D . Further studies on the blockade of 5-HT depolarizations of rabbit vagal afferent and sympathetic ganglion cells by MDL 72222 and other antagonists. Neuropharmacology. 1987; 26(1):39-48. DOI: 10.1016/0028-3908(87)90042-6. View

Kilpatrick G, Jones B, Tyers M . Identification and distribution of 5-HT3 receptors in rat brain using radioligand binding. Nature. 1987; 330(6150):746-8. DOI: 10.1038/330746a0. View

Fozard J . MDL 72222: a potent and highly selective antagonist at neuronal 5-hydroxytryptamine receptors. Naunyn Schmiedebergs Arch Pharmacol. 1984; 326(1):36-44. DOI: 10.1007/BF00518776. View

Ireland S, Tyers M . Pharmacological characterization of 5-hydroxytryptamine-induced depolarization of the rat isolated vagus nerve. Br J Pharmacol. 1987; 90(1):229-38. PMC: 1917290. DOI: 10.1111/j.1476-5381.1987.tb16844.x. View

Neijt H, te Duits I, Vijverberg H . Pharmacological characterization of serotonin 5-HT3 receptor-mediated electrical response in cultured mouse neuroblastoma cells. Neuropharmacology. 1988; 27(3):301-7. DOI: 10.1016/0028-3908(88)90048-2. View

Round A, WALLIS D . The depolarizing action of 5-hydroxytryptamine on rabbit vagal afferent and sympathetic neurones in vitro and its selective blockade by ICS 205-930. Br J Pharmacol. 1986; 88(2):485-94. PMC: 1916829. DOI: 10.1111/j.1476-5381.1986.tb10227.x. View

Rodbard D . Statistical quality control and routine data processing for radioimmunoassays and immunoradiometric assays. Clin Chem. 1974; 20(10):1255-70. View

Hagan R, Butler A, Hill J, Jordan C, Ireland S, Tyers M . Effect of the 5-HT3 receptor antagonist, GR38032F, on responses to injection of a neurokinin agonist into the ventral tegmental area of the rat brain. Eur J Pharmacol. 1987; 138(2):303-5. DOI: 10.1016/0014-2999(87)90450-x. View

10.

Hoyer D, Neijt H . Identification of serotonin 5-HT3 recognition sites in membranes of N1E-115 neuroblastoma cells by radioligand binding. Mol Pharmacol. 1988; 33(3):303-9. View

11.

Bradley P, Engel G, Feniuk W, Fozard J, Humphrey P, Middlemiss D . Proposals for the classification and nomenclature of functional receptors for 5-hydroxytryptamine. Neuropharmacology. 1986; 25(6):563-76. DOI: 10.1016/0028-3908(86)90207-8. View

12.

Richardson B, Engel G, Donatsch P, Stadler P . Identification of serotonin M-receptor subtypes and their specific blockade by a new class of drugs. Nature. 1985; 316(6024):126-31. DOI: 10.1038/316126a0. View

13.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

14.

Fozard J . Neuronal 5-HT receptors in the periphery. Neuropharmacology. 1984; 23(12B):1473-86. DOI: 10.1016/0028-3908(84)90091-1. View

15.

Amano T, Richelson E, Nirenberg M . Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc Natl Acad Sci U S A. 1972; 69(1):258-63. PMC: 427587. DOI: 10.1073/pnas.69.1.258. View

16.

Neijt H, Vijverberg H, van den Bercken J . The dopamine response in mouse neuroblastoma cells is mediated by serotonin 5HT3 receptors. Eur J Pharmacol. 1986; 127(3):271-4. DOI: 10.1016/0014-2999(86)90374-2. View