Characterisation of the Melatonin Receptor 1A (MTNR1A) gene in the Rasa Aragonesa sheep breed: association with reproductive seasonality.

The ovine Melatonin Receptor 1A (MTNR1A) gene was structurally characterised and association between its variants and the reproductive seasonality was examined in a daughter design comprising three families of Rasa Aragonesa sheep breed. Sequencing of six Rasa Aragonesa ewes with extreme values for seasonality trait revealed 28 polymorphisms: 11 SNPs in the coding region (all in Exon 2), and 17 SNPs in the promoter region MTNR1A. All the substitutions in the coding region were found most likely lacking any phenotypic effect, because they are conservative mutations or were not part of the transmembrane domain. The silent mutations, which had shown association with reproductive seasonality in other breeds, were also found and genotyped in Rasa Aragonesa. The T allele of SNP606/RsaI of MNTR1A gene was associated with a greater percentage of oestrous cyclic ewes in the Rasa Aragonesa breed, indicating that this SNP may be in linkage disequilibrium with a mutation responsible for this trait close to MTNR1A, or in regulatory sequences of the gene. In this sense, several SNPs affecting a binding element for some transcription factors have been identified in the promoter region. The SNPs at 422 and 527 positions could constitute a binding element for some transcription factors (TFs), located in an EF2 and SRY consensus sites in the promoter region, respectively. Haplotype h(5) showed significant differences with the h(2) haplotype (66% compared to 49.2%) on oestrous cyclicity, thus these results are consistent with genotypic associations for each SNP. Haplotype with T, A and T alleles for SNPs 422, 677 (promoter region) and 612 (Exon 2) showed an increase of the percentage of oestrous cyclic ewes. Although some of these mutations have been associated with seasonal reproduction, further studies with a more appropriate animal design as well as functional studies of TF binding activity are needed.