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[3H]cocaine labels a binding site associated with the serotonin transporter in guinea pig brain: Allosteric modulation by paroxetine

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Abstract

We studied the characteristics of [3H]cocaine binding to membranes prepared from whole guinea pig brain. Cocaine binding was specific and saturable. A one-site binding model fit the data adequately: the Kd value of [3H]cocaine was 44 nM with a Bmax value of 280 fmol/mg protein. The rank order of potency for the [3H]cocaine binding site was paroxetine > clomipramine > (−)-cocaine > fluoxetine > mazindol > desipramine > GBR12909 > phencyclidine > benztropine > GBR12935 > (+)-cocaine. The IC50 values of these drugs for inhibition of [3H]cocaine binding were highly correlated with their IC50 values for inhibition of [3H]5-HT uptake into synaptosomes prepared from whole guinea pig brain. High affinity 5-HT uptake inhibitors produced dose-dependent wash-resistant (pseudoirreversible) inhibition of [3H]cocaine binding. The wash-resistant inhibition produced by paroxetine was due to an increase in the Kd of [3H]cocaine binding sites, and was accompanied by an increase in the dissociation rate, consistent with an allosteric mechanism. These studies suggest that, using membranes prepared from whole guinea pig brain, [3H]cocaine labels a binding site associated with serotonin transporter and that paroxetine and cocaine bind to different sites on the serotonin transporter.

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Abbreviations

GBR12909:

1-(2-{bis(4-fluorophenyl)methoxy}ethyl)-4-{3-phenylpropyl}piperazine

TCP:

1-{1-(2-thienyl)cyclohexyl}piperidine

BTCP:

N-{1-(2-benzo(b)thiophenyl)cyclohexyl}piperidine

PCP:

1-(1-phenylcyclohexyl)piperidine

GBR12935:

(1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine)

CMI:

clomipramine

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Authors and Affiliations

  1. Laboratory of Clinical Psychopharmacology, NIDA Addiction Research Center, PO BOX 5180, 21224, Baltimore, MD

    Hyacinth C. Akunne & Richard B. Rothman

  2. Section on Drug Design and Synthesis, Laboratory of Medicinal Chemistry, NIDDK, 20892, Bethesda, MD

    Brian R. de Costa, Arthur E. Jacobson & Kenner C. Rice

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  1. Hyacinth C. Akunne
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  2. Brian R. de Costa
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  3. Arthur E. Jacobson
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  4. Kenner C. Rice
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  5. Richard B. Rothman
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Akunne, H.C., de Costa, B.R., Jacobson, A.E. et al. [3H]cocaine labels a binding site associated with the serotonin transporter in guinea pig brain: Allosteric modulation by paroxetine. Neurochem Res 17, 1275–1283 (1992). https://doi.org/10.1007/BF00968412

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