Characterization and expression of androgen receptors in olive flounder

Abstract

Androgens are critical hormones that regulate sex differentiation, sexual maturation, and spermatogenesis in vertebrates, which is mainly mediated by androgen receptors (ARs). Reports on transcript variants of ar (AR encoding gene) in human are almost always associated with cancers and androgen insensitivity syndrome. However, the knowledge of ar variants in teleosts is scarce. In this study, arβ and two transcript variants of arα (arα1 and arα2) in olive flounder (Paralichthys olivaceus) were cloned and analyzed. Their expression patterns were investigated in 16 adult female and male tissues by RT-PCR, respectively. arα1 was expressed in the majority of tissues excluding male liver, medulla oblongata and female cerebellum, with higher levels in male gonad, kidney, head kidney, intestine, stomach, spleen, heart and gill than in female. arα2 had similar expression patterns as arα1, with lower levels in general. arβ was also widely expressed in various tissues excluding male spleen, female spleen and gill, with higher levels in male gonad, kidney, head kidney, intestine and lower levels in hypothalamus than in female. Compared with arβ, much lower expression levels of arα1 and arα2 were detected in different brain areas. The real-time quantitative PCR (qPCR) results showed that the total arα expression level was relatively higher during olive flounder gonadal differentiation and before the onset of testis differentiation, whereas arβ was expressed significantly higher during male gonadal differentiation period than female gonadal differentiation period. The in vitro transient transfection assays showed that ARα1, ARα2 and ARβ could all suppress the activity of cyp19a (p450arom aromatase gene) promoter, and the inhibitory effect of ARα1 was dose dependent. Our results imply that arα1, arα2 and arβ are sex-related genes and they might play important roles in gonadal differentiation in flounder.

Keywords: Androgen receptor; Gene expression; Gene regulation; Gonadal differentiation; Paralichthys olivaceus; Transcript variant.