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Labelling of 5-Hydroxytryptamine3 Receptors with a Novel 5-HT3 Receptor Ligand, [3H]RS-42358–197
Erik H. F. Wong,
Douglas W. Bonhaus,
Irene Wu,
Eric Stefanich,
Richard M. Eglen,
Corresponding Author
Erik H. F. Wong
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Address correspondence and reprint requests to Dr. E. H. F. Wong at Institute of Pharmacology, Syntex Research, 3401 Hillview Ave., Palo Alto, CA 94304, U.S.A.Search for more papers by this authorDouglas W. Bonhaus
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorIrene Wu
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorEric Stefanich
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorRichard M. Eglen
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorErik H. F. Wong,
Douglas W. Bonhaus,
Irene Wu,
Eric Stefanich,
Richard M. Eglen,
Corresponding Author
Erik H. F. Wong
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Address correspondence and reprint requests to Dr. E. H. F. Wong at Institute of Pharmacology, Syntex Research, 3401 Hillview Ave., Palo Alto, CA 94304, U.S.A.Search for more papers by this authorDouglas W. Bonhaus
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorIrene Wu
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorEric Stefanich
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorRichard M. Eglen
Institute of Pharmacology, Syntex Research, Palo Alto, California, U.S.A.
Search for more papers by this authorAbstract
Abstract: RS-42358–197{(S)-N-(1-azabicyclo[2.2.2]oct-3-yl)-2,4,5,6-tetrahydro-1H-benzo[de]isoquinolin-1-one hydrochloride} displaced the prototypic 5-hydroxytryptamine3 (5-HT3) receptor ligand [3H]quipazine in rat cerebral cortical membranes with an affinity (pKi) of 9.8 ± 0.1, while having weak affinity (pKi < 6.0) in 23 other receptor binding assays. [3H]RS-42358–197 was then utilized to label 5-HT3 receptors in a variety of tissues. [3H]RS-42358–197 labelled high-affinity and saturable binding sites in membranes from rat cortex, NG108–15 cells, and rabbit ileal myenteric plexus with affinities (KD) of 0.12 ± 0.01, 0.20 ± 0.01, and 0.10 ± 0.01 nM and densities (Bmax) of 16.0 ± 2.0, 660 ± 74, and 88 ± 12 fmol/mg of protein, respectively. The density of sites labelled in each of these tissues with [3H]RS-42358–197 was similar to that labelled with [3H]GR 65630, but was significantly less than that found with [3H]-quipazine. The binding of [3H]RS-42358–197 had a pharmacological profile similar to that of [3H]quipazine, as indicated by the rank order of displacement potencies: RS-42358–197 > (S)-zacopride > tropisetron > (R)-zacopride > ondansetron > MDL72222 > 5-HT. However, differences in 5-HT3 receptors of different tissues and species were detected on the basis of statistically significant differences in the affinities of phenylbiguanide, and 1-(m-chlorophenyl)biguanide when displacing [3H]RS-42358-197 binding. [3H]RS-42358–197 also labelled a population (Bmax= 91 ± 17 fmol/mg of protein) of binding sites in guinea pig myenteric plexus membranes, with lower affinity (KD= 1.6 ± 0.3 nM) than those in the other preparations. Moreover, the rank order of displacement potencies of 15 5-HT3 receptor ligands in guinea pig ileum was found not to be identical to that in other tissues. Binding studies carried out with [3H]RS-42358–197 have detected differences in 5-HT3 receptor binding sites in tissues of different species and further underscore the unique nature of the guinea pig 5-HT3 receptor.
Abbreviations
-
- 5-HT
-
- 5-hydroxytryptamine (serotonin)
-
- mCPBG
-
- 1-(m-chlorophenyl)biguanide
-
- MK-801
-
- (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate
-
- NMDA
-
- N-methyl-d-aspartate
-
- RS-42358–197
-
- (S)-N-(1-azabicyclo[2.2.2]oct-3-yl)-2,4,5,6-tetrahydro-1H-benzo[de]isoquinolin-1-one hydrochloride
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