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Vol. 60, Issue 4, 632-639, October 2001
Institut de Recherches Servier, Division de Pharmacologie
Moléculaire et Cellulaire, Croissy sur Seine, France
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.
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