Targeted Disruption of the Mouse Mel_1b Melatonin Receptor

Abstract

Two high-affinity, G protein-coupled melatonin receptor subtypes have been identified in mammals. Targeted disruption of the Mel_1a melatonin receptor prevents some, but not all, responses to the hormone, suggesting functional redundancy among receptor subtypes (Liu et al., Neuron 19:91-102, 1997). In the present work, the mouse Mel_1b melatonin receptor cDNA was isolated and characterized, and the gene has been disrupted. The cDNA encodes a receptor with high affinity for melatonin and a pharmacological profile consistent with its assignment as encoding a melatonin receptor. Mice with targeted disruption of the Mel_1b receptor have no obvious circadian phenotype. Melatonin suppressed multiunit electrical activity in the suprachiasmatic nucleus (SCN) in Mel_1b receptor-deficient mice as effectively as in wild-type controls. The neuropeptide, pituitary adenylyl cyclase activating peptide, increases the level of phosphorylated cyclic AMP response element binding protein (CREB) in SCN slices, and melatonin reduces this effect. The Mel_1a receptor subtype mediates this inhibitory response at moderate ligand concentrations (1 nM). A residual response apparent in Mel_1a receptor-deficient C3H mice at higher melatonin concentrations (100 nM) is absent in Mel_1a-Mel_1b double-mutant mice, indicating that the Mel_1b receptor mediates this effect of melatonin. These data indicate that there is a limited functional redundancy between the receptor subtypes in the SCN. Mice with targeted disruption of melatonin receptor subtypes will allow molecular dissection of other melatonin receptor-mediated responses.

ACKNOWLEDGMENTS

We thank Chen Liu, Donald Hodges, and Stefanie Rassnick for assistance with preliminary studies and Camala Capodice and Christopher Lambert for technical assistance.

This work was supported by grants from the NIH (DK42125 to S.M.R. and AG09975 to D.R.W)., the Deutsche Forschungsgemeinschaft (to J.H.S and C.V.G.), the Paul und Ursula Klein-Stiftung and the Heinrich und Fritz Reise-Stiftung (to J.H.S), and a sponsored research agreement from Bristol-Myers Squibb. X.J. was supported in part by NIH Postdoctoral fellowship F32 MH12067. C.V.G. was supported in part by the Emmy-Noether Programm of the Deutsche Forschungsgemeinschaft.