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3- and 4-O-sulfoconjugated and methylated dopamine: highly reduced binding affinity to dopamine D2 receptors in rat striatal membranes

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Summary

The binding properties of 3- and 4-O-sulfoconjugated dopamine (DA-3-0-S, DA-4-0-S) as well as 3-O-methylated dopamine (MT) to rat striatal dopamine D2 receptors were investigated. 3H-spiperone was used as a radioligand in the binding studies. In saturation binding experiments (+)butaclamol, which has been reported to bind to dopaminergic D2 and serotoninergic 5HT2 receptors, was used in conjunction with ketanserin and sulpiride, which preferentially label 5HT2 and D2 receptors, respectively, in order to discriminate between 3H-spiperone binding to D2 and to 5HT2 receptors. Under our particular membrane preparation and assay conditions, 3H-spiperone binds to D2 and 5HT2 receptors with a maximal binding capacity (B max) of 340 fmol/mg protein in proportions of about 75%:25% with similar dissociation constants K D (35 pmol/l; 43 pmol/l). This result was verified by the biphasic competition curve of ketanserin, which revealed about 20% high (K D = 24 nmol/l) and 80% low (K D = 420 nmol/l) affinity binding sites corresponding to 5HT2 and D2 receptors, respectively. Therefore, all further competition experiments at a tracer concentration of 50 pmol/l were performed in the presence of 0.1 μmol/l ketanserin to mask the 5HT2 receptors. DA competition curves were best fitted assuming two binding sites, with high (K H = 0.12 μmol/l) and low (KL = 18 μmol/l) affinity, present in a ratio of 3:1. The high affinity binding sites were interconvertible by 100 μmol/l guanyl-5-yl imidodiphosphate [Gpp(NH)p], resulting in a homogenous affinity state of DA receptors (K D = 2.8 μmol/l). Competition experiments with various compounds confirmed the binding of 3H-spiperone to D2 receptors. DA-3-O-S, DA-4-O-S, and MT were more than 5,000-, more than 10,000-, and 530-fold less potent in competing for 3H-spiperone binding when compared with DA at the high affinity binding site which mediates biological effects. Therefore, it is concluded that these DA metabolites are biologically ineffective at central D2 receptors.

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Authors and Affiliations

  1. Department of Pathophysiology and Sportsmedicine, Medical Clinic and Policlinic, University of Heidelberg, Hospitalstrasse 3, D-6900, Heidelberg, Federal Republic of Germany

    Egon Werle, Theo Lenz, Günther Strobel & Helmut Weicker

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  1. Egon Werle
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  2. Theo Lenz
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  3. Günther Strobel
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  4. Helmut Weicker
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Werle, E., Lenz, T., Strobel, G. et al. 3- and 4-O-sulfoconjugated and methylated dopamine: highly reduced binding affinity to dopamine D2 receptors in rat striatal membranes. Naunyn-Schmiedeberg's Arch Pharmacol 338, 28–34 (1988). https://doi.org/10.1007/BF00168808

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