Genotype of MTNR1A gene regulates the conception rate following melatonin treatment in water buffalo.

Buffaloes have tendency to show seasonal reproduction and remain in anestrus due to limited ovarian activity during summer. The seasonal reproductive behavior is ascribed the effect of melatonin related to photoperiod. Treating animals with melatonin could be a possible strategy to overcome the problem. The role of MTNR1A gene has not been fully explained in the buffalo. Therefore, we conducted a study on 114 buffalo heifers to detect the polymorphic site in MTNR1A gene and further treated them with melatonin implants to investigate the role of most frequent genotype following melatonin treatment on pregnancy. The present investigation is the first to investigate the association between melatonin treated different MTNR1A genotype buffalo and pregnancy. We confirmed SNP at position 72 in 812 bp fragment exon II of MTNR1A gene. RFLP of PCR products with Hpa I enzyme resulted in three genotypes: TT (812bp), CT (812, 743, 69bp) and CC (743, 69bp). Next, buffaloes of each genotype (TT, CC, CT; n = 28 for each) were treated with melatonin implants to compare the conception rate with their corresponding untreated control (n = 10 for each genotype). Melatonin concentrations were higher (P < 0.05) for the treatment groups of all genotypes compared to their respective untreated control from day 1-28. The pregnancy rate was significantly associated with the MTNR1A genotype. The conception rate was higher (P < 0.05) for TT genotype than for the other genotypes of buffaloes treated with melatonin. Furthermore, buffaloes of TT genotype treated with melatonin started exhibiting estrus activity soon from second week of melatonin treatment (14.1 ± 2.1; range: 10-17 days) and were found to be 7.8 times more likely to become pregnant compared to other genotypes following melatonin treatment. In conclusion, TT genotype of MTNR1A gene is more sensitive to melatonin treatment that favours pregnancy in buffaloes during summer.