Characterization of binding sites for 3H-spiroperidol in human retina

P. McGonigle, M. B. Wax, P. B. Molinoff

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Binding sites for the D-2-selective antagonist (3H)-spiroperidol were characterized in human retina. Nonspecific binding, measured in the presence of 2 μM (+)-butaclamol, made up 20% of total binding. Scatchard analysis of the binding of (3H)-spiroperidol resulted in linear plots and yielded a K(d) value of 87 pM and a B(max) value of 1500 fmol/mg protein. In studies of the inhibition of the binding of (3H)-spiroperidol, (+)-butaclamol was approximately 1000-fold more potent than the (-)-stereoisomer. The inhibition curve for dopamine was shifted to the right and the Hill coefficient was increased by the addition of 300 μM GTP. This effect was agonist-specific and suggests that some of the receptors are coupled to stimulation or inhibition of the enzyme adenylate cyclase. The inhibition curves for most of the antagonists had Hill coefficients between 0.6 and 0.8. Hill coefficients were also consistently less than 1.0 for agonists even in the presence of GTP. Nonlinear regression analysis of untransformed data revealed that these shallow inhibition curves were best explained by the presence of two populations of binding sites, 40% of the sites having a high affinity for dopamine in the presence of GTP and domperidone and the remaining 60% having a lower affinity for these ligands. The larger population of sites had a higher affinity for sulpiride, fluphenazine, and N-propylnorapomorphine in the presence of GTP. The possibility that either of these classes of sites consisted of serotonin receptors was ruled out by the finding that the 5-HT2 antagonist ketanserin had a low affinity for both classes of sites.

Original languageEnglish (US)
Pages (from-to)687-694
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume29
Issue number5
StatePublished - 1988

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Spiperone
Guanosine Triphosphate
Retina
Butaclamol
Binding Sites
Dopamine
Domperidone
Fluphenazine
Serotonin 5-HT2 Receptor Antagonists
Ketanserin
Sulpiride
Stereoisomerism
Serotonin Receptors
Adenylyl Cyclases
Population
Regression Analysis
Ligands
Enzymes
Proteins

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Characterization of binding sites for 3H-spiroperidol in human retina. / McGonigle, P.; Wax, M. B.; Molinoff, P. B.

In: Investigative Ophthalmology and Visual Science, Vol. 29, No. 5, 1988, p. 687-694.

Research output: Contribution to journalArticle

McGonigle, P, Wax, MB & Molinoff, PB 1988, 'Characterization of binding sites for 3H-spiroperidol in human retina', Investigative Ophthalmology and Visual Science, vol. 29, no. 5, pp. 687-694.
McGonigle, P. ; Wax, M. B. ; Molinoff, P. B. / Characterization of binding sites for 3H-spiroperidol in human retina. In: Investigative Ophthalmology and Visual Science. 1988 ; Vol. 29, No. 5. pp. 687-694.
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