30The circadian and seasonal actions of melatonin are mediated by high affinity G-31 protein coupled receptors (melatonin receptors, MTRs), classified into 32 phylogenetically distinct subtypes based on sequence divergence and 33 pharmacological characteristics. Three vertebrate MTR subtypes are currently 34 described: MT1 (MTNR1A), MT2 (MTNR1B), and Mel1c (MTNR1C / GPR50), which 35 exhibit distinct affinities, tissue distributions and signaling properties. We present 36 phylogenetic and comparative genomic analyses supporting a revised classification 37 of the vertebrate MTR family. We demonstrate four ancestral vertebrate MTRs, 38including a novel molecule hereafter named Mel1d. We reconstructed the evolution 39 of each vertebrate MTR, detailing genetic losses in addition to gains resulting from 40 whole genome duplication events in teleost fishes. We show that Mel1d was lost 41 separately in mammals and birds and has been previously mistaken for an MT1 42 paralogue. The genetic and functional diversity of vertebrate MTRs is more complex 43 than appreciated, with implications for our understanding of melatonin actions in 44 different taxa. The significance of our findings, including the existence of Mel1d, are 45 discussed in an evolutionary and functional context accommodating a robust 46 phylogenetic assignment of MTR gene family structure. 47 50
51Melatonin is an ancient eukaryotic signalling molecule that regulates diverse 52 biological functions. While best known as a regulator of biological rhythms in 53 humans, this hormone also regulates energy balance, temperature, behavior, blood 54 pressure, and seasonal reproduction. Melatonin is secreted by the pineal gland and 55 targets the brain as well as peripheral tissues (Hardeland et al. 2011, Slominski et 56 al. 2012), but is also produced by several tissues, eliciting paracrine effects (Weaver 57 and Reppert 1990). The actions of melatonin depend on the spatiotemporal 58 expression of high-affinity melatonin receptors (MTR), representing a specific class of 59 G protein-coupled receptor (GPCR). 60 61