Immunohistochemical detection and biological activities of CYP17 (P450c17) in the indifferent gonad of the frog Rana rugosa

Nana Sakurai, Koichi Maruo, Shogo Haraguchi, Yoshinobu Uno, Yuki Oshima, Kazuyoshi Tsutsui, Yoichi Matsuda, Jean Luc Do Rego, Georges Pelletier, Hubert Vaudry, Masahisa Nakamura

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    Abstract

    Sex steroids play a crucial role in the gonad differentiation in various species of vertebrates. However, little is known regarding the localization and biological activity of steroid-metabolizing enzymes during gonadal sex differentiation in amphibians. In the present study, we showed by real-time RT-PCR analysis that the expression of CYP17, one of the key steroidogenic enzymes, was higher in the indifferent gonad during sex differentiation in male than in female tadpoles of Rana rugosa but that there was no difference detected in the 3βHSD mRNA level between the male and female gonads. We next examined the localization of CYP17, 3βHSD and 17βHSD in the indifferent and differentiating gonads by using three kinds of antibodies specific for CYP17, 3βHSD and 17βHSD, respectively. Positive signals for CYP17, 3βHSD and 17βHSD were observed in somatic cells of the indifferent gonad of males and in the interstitial cell of the testis. The enzymatic activity of CYP17 was also examined in the gonad during sex differentiation in this species. [3H]Progesterone (Prog) was converted to [3H]androstenedione (AE) in the indifferent gonad in males and females, but the rate of its conversion was higher in males than in females. Moreover, fluorescence in situ hybridization (FISH) analysis revealed that the CYP17 gene was located on the q arm of chromosome 9, indicating that CYP17 was autosomal in R. rugosa. Taken together, the results demonstrate that the CYP17 protein is synthesized in somatic cells of the indifferent gonad during gonadal sex differentiation in R. rugosa and that it is more active in converting Prog to AE in males than in females. The data suggest that CYP17 may be involved in testicular formation during sex differentiation in this species.

    Original languageEnglish
    Pages (from-to)5-12
    Number of pages8
    JournalJournal of Steroid Biochemistry and Molecular Biology
    Volume112
    Issue number1-3
    DOIs
    Publication statusPublished - 2008 Nov

    Keywords

    • CYP17
    • Enzymatic activity
    • FISH
    • Frog
    • Gonad
    • Immunohistochemistry
    • Real-time RT-PCR
    • Sex differentiation

    ASJC Scopus subject areas

    • Molecular Medicine
    • Endocrinology, Diabetes and Metabolism
    • Molecular Biology
    • Cell Biology
    • Endocrinology
    • Clinical Biochemistry
    • Biochemistry

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  • Cite this

    Sakurai, N., Maruo, K., Haraguchi, S., Uno, Y., Oshima, Y., Tsutsui, K., Matsuda, Y., Rego, J. L. D., Pelletier, G., Vaudry, H., & Nakamura, M. (2008). Immunohistochemical detection and biological activities of CYP17 (P450c17) in the indifferent gonad of the frog Rana rugosa. Journal of Steroid Biochemistry and Molecular Biology, 112(1-3), 5-12. https://doi.org/10.1016/j.jsbmb.2008.07.002