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

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

doi:10.1016/j.jsbmb.2008.07.002

Cited by 40 publications

(47 citation statements)

References 49 publications

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“…Therefore, in S. tropicalis , cyp19 lev-els are differentially expressed before the first signs of ovarian differentiation at NF51. In support of this observation, sexually dimorphic levels of GMC cyp19 in G. rugosa have been reported 2 weeks earlier than cyp17 and foxl2 [Ohtani et al, 2003;Iwade et al, 2008;Maruo et al, 2008;Oshima et al, 2008Oshima et al, , 2009Sakurai et al, 2008;Suda et al, 2011b]. Together, these results support the use of cyp19 as an early ovarian differentiation marker in amphibians, as has been proposed for fish [Fernandino et al, 2008;Blázquez et al, 2009].…”

Section: Development Of a Useful Tool To Determine Phenotypic Sex In supporting

confidence: 81%

“…Similarly, foxl2 expression was found higher in the GMCs of developing genetic females than in genetic males of G. rugosa tadpoles . In contrast, adult males, sex reversed male tadpoles and differentiating genetic male tadpoles express significantly higher gonadal levels of cyp17 than adult females or differentiating genetic female of G. rugosa tadpoles [Iwade et al, 2008;Maruo et al, 2008;Sakurai et al, 2008;Suda et al, 2011b]. Although genetic sex markers have not been established for many species, including L. sylvaticus , we can conclude that these 3 genes are good candidates for studying sex differentiation in frogs.…”

Section: Gender-specific Patterns Of Cyp19 Foxl2 and Cyp17 Can Be Obmentioning

confidence: 79%

“…The majority of these have focused on a single species, Glandirana (Rana) rugosa [Takase et al, 2000a, b;Sugita et al, 2001;Shibata et al, 2002;Aoyama et al, 2003;Ohtani et al, 2003;Oshima et al, 2005Oshima et al, , 2006Oshima et al, , 2007Oshima et al, , 2008Oshima et al, , 2009Yamamura et al, 2005;Iwade et al, 2008;Maruo et al, 2008;Nakamura et al, 2008;Sakurai et al, 2008;Yokoyama et al, 2009;Isomura et al, 2011;Suda et al, 2011a, b]. In this frog species, both genetic and phenotypic sex can be determined [Miura et al, 1998], and gene expression levels in differentiating GMCs can be studied separately in genetic males and females.…”

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confidence: 99%

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Sexing Frogs by Real-Time PCR: Using Aromatase (cyp19) as an Early Ovarian Differentiation Marker

Navarro‐Martín

Velasco-Santamaría²,

Duarte‐Guterman³

et al. 2012

Sex Dev

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Most anurans have no identified sex-markers; therefore, alternative methods for identification of early changes in sex ratios are required. In this study, Lithobates sylvaticus and Silurana tropicalis tadpoles were sampled at different developmental stages covering the entire process of sex differentiation. Three candidate genes known to be involved in sex differentiation in other vertebrate species were selected to develop a method to identify phenotypic sex in frogs: cytochrome p450 aromatase (cyp19), forkhead box L2 (foxl2) and the cytochrome 17-alpha-hydroxylase/17,20 lyase (cyp17). Cloning of these genes revealed nucleotide identity values ranging between 75–97% when compared to other amphibian species. Gene expression of cyp17,cyp19 and foxl2 in L. sylvaticus adult gonads and gonad-mesonephros complex (GMC) of tadpoles was analyzed by real-time RT-PCR. Results showed clear sexually dimorphic patterns in the expression of the 3 genes. Our analysis reveals that GMC gene expression levels of cyp19 alone can be used as a robust predictor of phenotypic sex in L. sylvaticus tadpoles. In addition, we validated this method measuring cyp19 mRNA levels in S. tropicalis GMCs. We propose measuring cyp19 as a tool to study the effects of chemical contaminants (including endocrine disrupting compounds) on amphibian gonadal development and sex ratios in the future.

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Section: Development Of a Useful Tool To Determine Phenotypic Sex In supporting

confidence: 81%

Section: Gender-specific Patterns Of Cyp19 Foxl2 and Cyp17 Can Be Obmentioning

confidence: 79%

mentioning

confidence: 99%

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Sexing Frogs by Real-Time PCR: Using Aromatase (cyp19) as an Early Ovarian Differentiation Marker

Navarro‐Martín

Velasco-Santamaría²,

Duarte‐Guterman³

et al. 2012

Sex Dev

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“…Recently, interesting results were published about R. rugosa . Three enzymes (CYP17, HSD3B, and HSD17B) were detected immunohistochemically in the indifferent gonads of male and female tadpoles during sex-determination [Sakurai et al, 2008]. When the enzymatic activity of CYP17 and HSD17B was measured by HPLC analysis, indifferent male gonads converted progesterone to testosterone via androstenedione at a 2-fold higher rate compared to the indifferent gonads of females [Sakurai et al, 2008].…”

Section: Activity Of Steroidogenic Enzymesmentioning

confidence: 99%

“…Three enzymes (CYP17, HSD3B, and HSD17B) were detected immunohistochemically in the indifferent gonads of male and female tadpoles during sex-determination [Sakurai et al, 2008]. When the enzymatic activity of CYP17 and HSD17B was measured by HPLC analysis, indifferent male gonads converted progesterone to testosterone via androstenedione at a 2-fold higher rate compared to the indifferent gonads of females [Sakurai et al, 2008]. Estradiol-17 ␤ production is higher in the indifferent gonad of females than in that of males during sexdetermination in R. rugosa [Isomura et al, 2011].…”

Section: Activity Of Steroidogenic Enzymesmentioning

confidence: 99%

Is a Sex-Determining Gene(s) Necessary for Sex-Determination in Amphibians? Steroid Hormones May Be the Key Factor

Nakamura¹

2012

Sex Dev

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Amphibians have 2 genetic sex-determining systems, one with male (XX/XY) and one with female (ZZ/ZW) heterogamety. While the ancestral state of sex-determination is thought to be female heterogamety, male and female heterogametic types were probably once interchangeable. The Japanese frog Rana rugosa has both XX/XY and ZZ/ZW systems within a single species in certain local populations. However, steroid hormones can alter the phenotypic sex epigenetically. In R. rugosa, steroidogenic enzyme expression starts before sex-determination in the indifferent gonad, and these enzymes become active in both male and female tadpoles. Androgens are produced in the indifferent gonad of male tadpoles at high levels, whereas estrogens are synthesized in females. In this regard, the observed enhanced expression of the hormone-metabolizing genes, CYP19 in the female gonad and CYP17 in males, may be crucial for sex-determination. Moreover, with FSH known to increase estrogen synthesis in the vertebrate ovary, observed upregulation of FSH receptor (FSHR) expression in the indifferent gonad of female tadpoles is intriguing. These data suggest that steroid hormones could be crucial for sex-determination in R. rugosa, with the consequence that upregulation of CYP19 and FSHR expression is necessary for female and CYP17 for male sex-determination.

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Development and steroidogenic properties of the Bidder's organ of the tadpole of Rhinella arenarum (Amphibia, Anura)

et al. 2014

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Several studies suggested that in anuran amphibians steroidogenic enzymes are critical for gonadal differentiation, proposing that the amount of sex steroids would adjust this differentiation. Among anurans, bufonids are important for the study of sex differentiation due to the presence of Bidder's organ (BO) that differentiates as a rudimentary ovary in the cephalic portion of the genital ridge. Considering that in adult males of Rhinella arenarum, the BO synthesizes estradiol, the main purpose of this work is to examine, in this species, the morphogenesis of BO and the steroidogenic capacity of this organ during larval development. BO and the proper gonads are distinguished from Gosner stage 26. During metamorphosis, BO primary oogonia develop in oogonia in nests, early previtellogenic oocytes and late previtellogenic oocytes in follicles while proper gonads remain undifferentiated. Aromatase was detected by immunohistochemistry in almost all the largest follicles of the BOs while the cytochrome P450 side-chain cleavage was observed in only few oocytes. The proper gonad was not immunoreactive in any stage. The determination of aromatase and 5α-reductase activities showed that the population of tadpoles between stages 36-41 is not homogeneous in terms of aromatase activity. In addition, from stage 26 to the end of metamorphosis, all the stages were able to produce estradiol from endogenous substrate but stages 40-41, corresponding to the end of pro-metamorphosis, produced the highest values. In conclusion, BO is able to synthesize estradiol from endogenous precursors and proper gonad remains undifferentiated at least until the end of the metamorphosis.

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

Cited by 40 publications

References 49 publications

Sexing Frogs by Real-Time PCR: Using Aromatase <b><i>(cyp19) </i></b>as an Early Ovarian Differentiation Marker

Sexing Frogs by Real-Time PCR: Using Aromatase <b><i>(cyp19) </i></b>as an Early Ovarian Differentiation Marker

Is a Sex-Determining Gene(s) Necessary for Sex-Determination in Amphibians? Steroid Hormones May Be the Key Factor

Development and steroidogenic properties of the Bidder's organ of the tadpole of Rhinella arenarum (Amphibia, Anura)

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