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Modification of human 5-HT2C receptor function by Cys23Ser, an abundant, naturally occurring amino-acid substitution

Molecular Psychiatry volume 9, pages 55–64 (2004)Cite this article

Abstract

A human serotonin (5-HT)2C receptor gene polymorphism leads to the substitution of cysteine for serine at codon 23 (Cys23Ser); the frequency of the Ser23 allele in unrelated Caucasians is approximately 0.13. In the present study, we assessed whether Cys23Ser could affect receptor function. The two alleles were functionally compared following expression in COS-7 cells. The constitutive activity of the receptor in an in situ reconstitution system was also evaluated following expression of each allele in Sf9 cells. Using radioligands, Ser23-expressed membranes showed reduced high-affinity binding to meta-chlorophenylpiperazine (m-CPP) and 5-HT. Although the amplitude of the 5-HT-induced intracellular Ca2+ peak did not differ between the alleles, Ser23 required higher 5-HT concentrations to elicit the same response. These differences might be due to more extensive desensitization in the Ser23 form. In the in situ reconstitution system, the 5-HT2C receptor displayed considerable constitutive activity, with the Ser23 allele being significantly higher in this regard than the Cys23 form. After prolonged serum deprivation in order to resensitize the receptor, four of the 15 cells expressing Ser23 showed abnormally higher m-CPP-induced sensitivity of the Ca2+ response. These results indicate that the Ser23 allele may be constitutively more active than Cys23. Thus, Ser23 appears to be an abundant candidate allele capable of directly influencing inter-individual variation in behavior, susceptibility to mental disorder, and response to drugs including atypical antipsychotic and some antidepressant drugs that are potent 5-HT2C inverse agonists or antagonists.

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Acknowledgements

Dr Alan G Saltzman at Rhone-Poulenc Rorer is gratefully acknowledged for providing us with the clone of the human 5-HT2C receptor. Many thanks to Mr Loren Chen, Dr Glenn Kroog, Dr Xiaoying Jian, and Dr Bill Clark (National Institute on Deafness and Other Communication Disorders, NIH) for assistance with purification of G proteins and helpful discussion. We also thank Professor Herbert Y Meltzer (Vanderbilt University School of Medicine) for a critical review of the manuscript. This work was partly supported by the Research Grant (10670923, 13470198, 13877152) from the Ministry of Science Education and Culture of Japan, the Research Grant for Nervous and Mental Disorders from the Ministry of Health and Welfare of Japan, and Special Coordination Funds for Promoting Science and Technology Target-oriented Brain Science Research Program from the Ministry of Science and Technology of Japan.

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Author notes

  1. M Linnoila: Deceased.

  2. M Okada: Previous address: Laboratory of Neurogenetics, NIAAA, NIH Clinical Center, Flow Bldg. Room 2, 12501 Washington Avenue, Rockville, MD 20852, USA.

Authors and Affiliations

  1. Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, MD, USA

    N Ozaki & D Goldman

  2. Department of Public Health, Faculty of Medicine, Osaka City University, Osaka, Japan

    M Okada

  3. Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Rockville, MD, USA

    J K Northup

  4. Laboratory of Cellular Molecular Neurophysiology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA

    J T Russell

  5. Laboratory Clinical Studies, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, MD, USA

    M Linnoila

  6. Department of Psychiatry, Fujita Health University, Toyoake, Japan

    N Ozaki

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Correspondence to M Okada.

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Okada, M., Northup, J., Ozaki, N. et al. Modification of human 5-HT2C receptor function by Cys23Ser, an abundant, naturally occurring amino-acid substitution. Mol Psychiatry 9, 55–64 (2004). https://doi.org/10.1038/sj.mp.4001357

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