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Molecular Pharmacology

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Rapid CommunicationAccelerated Communication

Cloning and Molecular Characterization of the Novel Human Melanin-Concentrating Hormone Receptor MCH2

M. Rodriguez, P. Beauverger, I. Naime, H. Rique, C. Ouvry, S. Souchaud, S. Dromaint, N. Nagel, T. Suply, V. Audinot, J. A. Boutin and J. P. Galizzi
Molecular Pharmacology October 2001, 60 (4) 632-639;
M. Rodriguez
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P. Beauverger
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I. Naime
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H. Rique
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C. Ouvry
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S. Souchaud
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S. Dromaint
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N. Nagel
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T. Suply
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V. Audinot
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J. A. Boutin
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Abstract

Using a genomics-based approach for screening orphan G-protein-coupled receptors, we have identified and cloned a novel high-affinity, melanin-concentrating hormone (MCH) receptor. This receptor, named S643b, displays the greatest overall identity (32%) with the previously reported human SLC-1 receptor (MCH1) and to a lesser extent with the somatostatin receptor subtypes. The gene encoding the S643b receptor spans more than 23 kilobase pairs (kb) and was mapped, by radiation hybrid experiments, on chromosome 6q14.3-q15. Comparison of the S643b cDNA with human genomic sequence reveals that the 340-amino-acid receptor is encoded by five exons. Its tissue distribution, as determined by Northern blot and reverse transcription-polymerase chain reaction analysis, indicates that a 4-kb transcript is predominantly expressed in the brain. When expressed in Chinese hamster ovary (CHO) cells, the S643b receptor displays a strong, dose-dependent, transient elevation of intracellular calcium in response to MCH (EC50 = 9.5 nM). During the present study, we isolated a splice variant, designated S643a, encoding for a receptor that was not activated by MCH in a cellular calcium mobilization assay. Comparative pharmacological studies using CHO cells stably expressing either SLC-1 or S643b receptors demonstrated that similar structural features of MCH are required to stimulate intracellular Ca2+ mobilization at both receptors. The identification and localization of this new MCH receptor (MCH2) provides further insight into the physiological implication of MCH in modulating behavioral responses, including food intake.

Footnotes

  • T.S. was the recipient of a Convention Industrielle de Formation par la Recherche between the Association Nationale de la Recherche Technique, the Institut de Recherche Servier, and the Centre National de la Recherche Scientifique.

  • Abbreviations:
    MCH
    melanin-concentrating hormone
    GPCR
    G-protein-coupled receptor
    RT
    reverse transcriptase
    PCR
    polymerase chain reaction
    bp
    base pair
    HTGS
    high throughput genome sequences
    RACE
    rapid amplification of cDNA ends
    SSC
    standard saline citrate
    CHO
    Chinese hamster ovary
    FLIPR
    fluorometric imaging plate reader
    HEK
    human embryonic kidney
    • Received June 15, 2001.
    • Accepted July 11, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 60 (4)
Molecular Pharmacology
Vol. 60, Issue 4
1 Oct 2001
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Rapid CommunicationAccelerated Communication

Cloning and Molecular Characterization of the Novel Human Melanin-Concentrating Hormone Receptor MCH2

M. Rodriguez, P. Beauverger, I. Naime, H. Rique, C. Ouvry, S. Souchaud, S. Dromaint, N. Nagel, T. Suply, V. Audinot, J. A. Boutin and J. P. Galizzi
Molecular Pharmacology October 1, 2001, 60 (4) 632-639;

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Rapid CommunicationAccelerated Communication

Cloning and Molecular Characterization of the Novel Human Melanin-Concentrating Hormone Receptor MCH2

M. Rodriguez, P. Beauverger, I. Naime, H. Rique, C. Ouvry, S. Souchaud, S. Dromaint, N. Nagel, T. Suply, V. Audinot, J. A. Boutin and J. P. Galizzi
Molecular Pharmacology October 1, 2001, 60 (4) 632-639;
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