Melatonin rhythms beyond the pineal organ: gene expression of receptors and biosynthesis enzymes in wild and F1 Senegalese sole.

Abstract

Fish gonadal melatonin production is still unexplored and could contribute to a better understanding of its role in reproduction control, especially for species with reproductive impairments. This study aimed to comprehend if Senegalese sole testes are an extra-pineal production site of melatonin and if it has seasonal and daily variations. Wild and F1 broodstocks were sampled in the breeding season (BS) and out of the reproductive season (OS), at mid-light (ML) and mid-dark (MD) daytimes. Blood plasma melatonin concentration was determined by radioimmunoassay (RIA). The expression of genes involved in melatonin biosynthesis (tph1a, tph2, hiomt1, aanat1a, aanat1b, and aanat2) and melatonin receptors (mel1, mel1c, and mel2) was evaluated in the brain, eye, and testis by quantitative real-time PCR (qPCR). Plasma melatonin concentration in wild sole displayed day/night differences in both seasons (average ML: 36 ± 22 pg/mL, MD: 108 ± 63 pg/mL), whereas differences in the F1 broodstock were only found OS (ML: 100 ± 54 pg/mL, MD: 187 ± 88 pg/mL). Gene expression of mel1 and mel2 receptors, and tph1a, aanat1a, aanat2, and hiomt1 enzymes was detected and quantified in the fish testes. Moreover, daily and seasonal fluctuations in the expression of those genes were found in all tissues and broodstock groups. However, the F1 group showed distinct gene expression patterns compared to the wild type, suggesting a disruption in the circadian system. This study revealed that Senegalese sole testes are a melatonin production site and, at the same time, suggested a dysregulation in the hypothalamus-pituitary-gonad (HPG) axis of F1 males.