Approaches to the purification of the 5-hydroxytryptamine reuptake system from human blood platelets

Biessen, E.A.L.; Horn, Alan S.; Robillard, George T.

doi:10.1042/bst0190103

Cited by 2 publications

(3 citation statements)

References 8 publications

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“…The 5-HT uptake system in platelets and neurons share many similarities (Segonzac et al, 1985) . However, there is no complete evidence on the identity of both systems (Biessen et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

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Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light

Lima

Schmeer

1994

Journal of Neurochemistry

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The serotonin (5‐HT) uptake system of goldfish retina was evaluated by the binding of [3H]paroxetine to membrane preparations and the uptake of [3H]5‐HT into isolated cells from goldfish retina. The order of potency of inhibitors of [3H]paroxetine binding was imipramine > 5‐methoxy‐N,N‐dimethyltryptamine > desipramine > fluoxetine > citalopram > 5‐HT. The saturation experiments indicated a high‐affinity binding site, and positive cooperativity with Hill coefficient higher than unity. The association reached equilibrium at about one hour of incubation and was efficiently displaced by imipramine. The equilibrium dissociation constants calculated by the antilog of the log concentration of ligand giving 50% of occupation, and by the ratio of dissociation/association constants, were similar: 5.84 and 2.34 nM, respectively. The binding was not significantly reduced by decreasing the temperature of incubation and was sodium dependent. The lesion with 5,7‐dihydroxytryptamine reduced the binding to 60%. The uptake of [3H]5‐HT into isolated cells also showed positive cooperativity. The order of potency of inhibitors was similar to the one obtained for the binding of [3H]paroxetine. Darkness increased the uptake of 5‐HT. The allosteric regulation of the 5‐HT transporter and the modulation by light could be related to the physiological role of the monoamine, as a neurotransmitter and as a precursor of melatonin synthesis in the retina.

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Section: Discussionmentioning

confidence: 99%

“…The 5-HT transport system in the CNS has been studied by classical methods (Snyder et al, 1970;Ross, 1987). This transport operates as a symporter and is sodium dependent and partially chloride dependent (Biessen et al, 1991). Recently, neurotransmitter transporters have been identified as a new gene family (Uhl, 1992;Uhl and Hartig, 1992).…”

mentioning

confidence: 99%

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light

Lima

Schmeer

1994

Journal of Neurochemistry

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“…Pharmacopsychiat. 27 (1994) 17 Biochemical purification has been hampered by the minute quantities of protein available and by the difficulty in working with membrane proteins (for example, see Biessen et al, 1991). Thus, there has been little molecular information on the Na+/CI--dependent monoamine transporters.…”

Section: Introductionmentioning

confidence: 99%

Expression Cloning of a Serotonin Transporter: A New Way to Study Antidepressant Drugs

Hoffman¹

1994

Pharmacopsychiatry

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Tricyclic antidepressants revolutionized the treatment of depression. These results and the monoamine-depleting effect of reserpine have contributed to the proposal that an imbalance in monoamines is a causal factor in depression. Most antidepressants act to concentrate monoamines in the synapse either by blocking metabolism via monoamine oxidase or by inhibiting reuptake by plasma membrane transporters. We have used a novel cDNA expression cloning strategy to isolate cDNAs for the antidepressant-sensitive serotonin transporter and for a reserpine-sensitive vesicular monoamine transporter which is critical for packaging serotonin, dopamine, norepinephrine, epinephrine and histamine into synaptic vesicles and secretory granules. In addition, we have isolated a dopamine transporter. The three papers summarized here describe the molecular characteristics of three proteins critical for monoamine neurotransmission and which are targets for antidepressant and psychostimulant drugs. The cloning of the serotonin and dopamine transporters and of the CNS vesicular transporter provide new tools to examine how post-translational and transcriptional regulation of these transporters effect uptake, storage and release of monoamines in normal and disease states. In addition to providing substrates for further drug discovery, isolation of human homologues should be useful for assessing the possible genetic bases of depression.

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Approaches to the purification of the 5-hydroxytryptamine reuptake system from human blood platelets

Cited by 2 publications

References 8 publications

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light

Expression Cloning of a Serotonin Transporter: A New Way to Study Antidepressant Drugs

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Approaches to the purification of the 5-hydroxytryptamine reuptake system from human blood platelets

Cited by 2 publications

References 8 publications

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [3H]Paroxetine and the Uptake of [3H]Serotonin: Modulation by Light

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [3H]Paroxetine and the Uptake of [3H]Serotonin: Modulation by Light

Expression Cloning of a Serotonin Transporter: A New Way to Study Antidepressant Drugs

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Product

Resources

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Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light

Characterization of Serotonin Transporter in Goldfish Retina by the Binding of [³H]Paroxetine and the Uptake of [³H]Serotonin: Modulation by Light