| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1991435 | The Journal of Steroid Biochemistry and Molecular Biology | 2014 | 10 Pages |
•Type 8 17β-HSD was cloned and characterized in scallop Chlamys farreri.•Recombinant 17β-HSD8 catalyzed steroids metabolism in vitro.•Variations of 17β-HSD8 expression levels were correlated with gametogenesis.•17β-HSD8 was located in follicle cells and gametes except spermatozoa.•17β-HSD8 may regulate estradiol-mediated gametogenesis in the gonad.
17β-Hydroxysteroid dehydrogenases (17β-HSDs) are important enzymes catalyzing steroids biosynthesis and metabolism in vertebrates. Although studies indicate steroids play a potential role in reproduction of molluscs, little is known about the presence and function of 17β-HSDs in molluscs. In the present study, a full-length cDNA encoding 17β-HSD type 8 (17β-HSD8) was identified in the Zhikong scallop Chlamys farreri, which is 1104 bp in length with an open reading frame of 759 bp encoding a protein of 252 amino acids. Phylogenetic analysis revealed that the C. farreri 17β-HSD8 (Cf-17β-HSD8) belongs to the short chain dehydrogenase/reductase family (SDR) and shares high homology with other 17β-HSD8 homologues. Catalytic activity assay in vitro demonstrated that the refolded Cf-17β-HSD8 expressed in Escherichia coli could effectively convert estradiol-17β (E2) to estrone (E1), and weakly catalyze the conversion of testosterone (T) to androstenedione (A) in the presence of NAD+. The Cf-17β-HSD8 mRNA was ubiquitously expressed in all tissues analyzed, including gonads. The expression levels of Cf-17β-HSD8 mRNA and protein increased with gametogenesis in both ovary and testis, and were significantly higher in testis than in ovary at growing stage and mature stage. Moreover, results of in situ hybridization and immunohistochemistry revealed that the mRNA and protein of Cf-17β-HSD8 were expressed in follicle cells and gametes at all stages except spermatozoa. Our findings suggest that Cf-17β-HSD8 may play an important role in regulating gametogenesis through modulating E2 levels in gonad of C. farreri.
