Characterization and expression of StAR2a and StAR2b in the olive flounder Paralichthys olivaceus

Steroidogenic acute regulatory protein 2 (StAR2) is a key protein in transporting cholesterol from the outer mitochondria membrane to the inner mitochondria membrane for sex steroid synthesis. In this study, two StAR2 gene isoforms, StAR2a and StAR2b, were isolated from the olive flounder Paralichthys olivaceus gonads. Semi-quantitative RT-PCR results indicated that their expression levels were higher in testis than those in ovary. StAR2a was mainly expressed in the thecal cells and ooplasm of ovary, and Leydig cells and spermatid of testis according to the results of in situ hybridization. The quantitative real-time PCR results showed that the expressions of StAR2a and StAR2b were high in undifferentiation gonads and differentiating testis, and then decreased in differentiated testis in the high temperature (28°C) and exogenous testosterone treatment groups. While, in the exogenous 17β-estradiol treatment group, both genes' expression levels were high in differentiating ovary, and then significantly decreased in differentiated ovary (P<0.05). StAR2a and StAR2b expression levels were significantly down-regulated in the cultured testis cells treated with the 75 and 150μM cAMP, but significantly up-regulated in the cultured testis cells treated with the 300μM cAMP (P<0.05). Moreover, their expression levels were significantly up-regulated by transfecting the cultured testis cells with pcDNA3.1-NR5a2 and pcDNA3.1-NR0b1 (P<0.05). Above study showed that expression of StAR2 was regulated by cAMP and the transcription factors, NR5a2 and NR0b1, indicating that StAR2 may have functions in flounder gonadal differentiation and maintenance.