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Cloning and Expression of a Human 5-HT1D Receptor

  • Chapter
Serotonin: Molecular Biology, Receptors and Functional Effects

Summary

A gene encoding a serotonin 5-HT1D receptor was cloned from a human brain cDNA library and functionally expressed in a heterologous expression system. Radioligand binding experiments were used to evaluate the pharmacological properties of membrane fragments expressing this cloned human receptor. [3H]5-HT bound with high affinity (Kd = 4.3 nM); the site density was 5.2 pmoles/mg protein. Competition studies revealed the following affinities (Ki in nM): 5-CT = 0.67; 5-HT = 4.2; 5-methoxytryptamine = 4.7; yohimbine = 25; spiperone > 1000; pindolol > 1000; zacopride > 10,000. High affinity binding was sensitive to the addition of guanine nucleotides, indicating functional coupling to a G-protein. 5-HT was found to inhibit forskolin-stimulated adenylate cyclase activity in L-M(TK-) cells expressing the cloned receptor.

These data are consistent with the assignment of the newly cloned gene as a 5-HT1D receptor. Since the deduced amino acid sequence of this gene exhibits only moderate homology to the 5-HT1A receptor and low homology to 5-HT2 receptor subfamily (5-HT2 and 5-HT1C receptors), the human 5-HT1D receptor apparently represents the first member of a new subfamily of serotonin receptor genes. Since subtypes of the 5-HT1D receptor have been hypothesized, this human 5-HT1D clone will facilitate the isolation of any additional 5-HT1D receptor subtypes in the human brain.

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Authors and Affiliations

  1. Neurogenetic Corporation, Paramus, 07652, NJ, USA

    T. Branchek, J. Zgombick, M. Macchi, P. Hartig & R. Weinshank

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  1. T. Branchek
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  2. J. Zgombick
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  3. M. Macchi
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  4. P. Hartig
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  5. R. Weinshank
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Editors and Affiliations

  1. Preclinical Research, Sandoz Pharma Ltd., P.O. Box 6, CH-4002, Basel, Switzerland

    John R. Fozard

  2. Institute of Pharmacology, Erasmus University Rotterdam, P.O. Box 1738, NL-3000 DR, Rotterdam, The Netherlands

    Pramod R. Saxena

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© 1991 Birkhäuser Verlag Basel/Switzerland

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Branchek, T., Zgombick, J., Macchi, M., Hartig, P., Weinshank, R. (1991). Cloning and Expression of a Human 5-HT1D Receptor. In: Fozard, J.R., Saxena, P.R. (eds) Serotonin: Molecular Biology, Receptors and Functional Effects. Birkhäuser Basel. https://doi.org/10.1007/978-3-0348-7259-1_3

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  • DOI: https://doi.org/10.1007/978-3-0348-7259-1_3

  • Publisher Name: Birkhäuser Basel

  • Print ISBN: 978-3-0348-7261-4

  • Online ISBN: 978-3-0348-7259-1

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