Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9

Yasutaka Yamamura, Sho Aoyama, Yuki Oshima, Takuma Kato, Noboru Osawa, Masahisa Nakamura

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    Sry (sex-determining region on the Y chromosome) is required for testicular differentiation in mammals. In addition to Sry, other genes such as WT1, Fgf9, Dax1, Dmrt1 and Sox9 are widely accepted to be involved in the sex determination in vertebrates. However, the roles of these genes during sex determination still remain unclear in amphibians. This study was undertaken to examine the expression of WT1 and Fgf9 in the developing gonad of amphibians. We first isolated the WT1 cDNA from the frog Rana rugosa. Like WT1 in mice, R. rugosa WT1 showed 2 isoforms; i.e., one had an additional 3 amino acids, KTS, included between the third and fourth zinc fingers. However, 17 amino acids in exon 5 of mammalian WT1 could not be found in R. rugosa WT1, which is also the case in turtle and chicken. The mRNA of both isoforms (+KTS, -KTS) was detected in the lung, kidney and testis, but not in the ovary and muscle of adult frogs. The 2 isoforms were expressed first in the embryos at stage 23. Thereafter, the expressions remained constant in the gonad attached to mesonephros of both sexes during sex determination. We next isolated the R. rugosa Fgf9 cDNA encoding 208 amino acids. The amino acid sequence of Fgf9 had similarity greater than 92% with chicken, mouse and human Fgf9s, suggesting that Fgf9 is highly conserved among vertebrate classes. Fgf9 was expressed in the ovary of an adult frog strongly, but in the lung weakly. In contrast, the Fgf9 mRNA was hardly detected in the kidney, testis and muscle. Moreover, Fgf9 did not show a sexually dimorphic expression pattern during sex determination in R. rugosa. The results, taken together, suggest that both WT1 and Fgf9 are expressed in the indifferent gonad prior to sex determination without any difference in the expression between males and females. Thus, it seems unlikely that they are a key factor to initiate the divergence leading to testicular or ovarian differentiation in R. rugosa.

    Original languageEnglish
    Pages (from-to)1045-1050
    Number of pages6
    JournalZoological Science
    Volume22
    Issue number9
    DOIs
    Publication statusPublished - 2005 Sep

    Fingerprint

    gonads
    molecular cloning
    gender
    frogs
    amino acids
    amphibians
    testes
    lungs
    vertebrates
    kidneys
    chickens
    muscles
    zinc finger motif
    mice
    Y chromosome
    Glandirana rugosa
    turtles
    exons
    embryo (animal)
    genes

    Keywords

    • cDNA
    • Fgf9
    • Gonad
    • Rana rugosa
    • RT-PCR
    • WT1

    ASJC Scopus subject areas

    • Animal Science and Zoology

    Cite this

    Yamamura, Y., Aoyama, S., Oshima, Y., Kato, T., Osawa, N., & Nakamura, M. (2005). Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9. Zoological Science, 22(9), 1045-1050. https://doi.org/10.2108/zsj.22.1045

    Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9. / Yamamura, Yasutaka; Aoyama, Sho; Oshima, Yuki; Kato, Takuma; Osawa, Noboru; Nakamura, Masahisa.

    In: Zoological Science, Vol. 22, No. 9, 09.2005, p. 1045-1050.

    Research output: Contribution to journalArticle

    Yamamura, Y, Aoyama, S, Oshima, Y, Kato, T, Osawa, N & Nakamura, M 2005, 'Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9', Zoological Science, vol. 22, no. 9, pp. 1045-1050. https://doi.org/10.2108/zsj.22.1045
    Yamamura Y, Aoyama S, Oshima Y, Kato T, Osawa N, Nakamura M. Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9. Zoological Science. 2005 Sep;22(9):1045-1050. https://doi.org/10.2108/zsj.22.1045
    Yamamura, Yasutaka ; Aoyama, Sho ; Oshima, Yuki ; Kato, Takuma ; Osawa, Noboru ; Nakamura, Masahisa. / Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9. In: Zoological Science. 2005 ; Vol. 22, No. 9. pp. 1045-1050.
    @article{d6bd24408c0b402db3ed7d9601b5781e,
    title = "Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9",
    abstract = "Sry (sex-determining region on the Y chromosome) is required for testicular differentiation in mammals. In addition to Sry, other genes such as WT1, Fgf9, Dax1, Dmrt1 and Sox9 are widely accepted to be involved in the sex determination in vertebrates. However, the roles of these genes during sex determination still remain unclear in amphibians. This study was undertaken to examine the expression of WT1 and Fgf9 in the developing gonad of amphibians. We first isolated the WT1 cDNA from the frog Rana rugosa. Like WT1 in mice, R. rugosa WT1 showed 2 isoforms; i.e., one had an additional 3 amino acids, KTS, included between the third and fourth zinc fingers. However, 17 amino acids in exon 5 of mammalian WT1 could not be found in R. rugosa WT1, which is also the case in turtle and chicken. The mRNA of both isoforms (+KTS, -KTS) was detected in the lung, kidney and testis, but not in the ovary and muscle of adult frogs. The 2 isoforms were expressed first in the embryos at stage 23. Thereafter, the expressions remained constant in the gonad attached to mesonephros of both sexes during sex determination. We next isolated the R. rugosa Fgf9 cDNA encoding 208 amino acids. The amino acid sequence of Fgf9 had similarity greater than 92{\%} with chicken, mouse and human Fgf9s, suggesting that Fgf9 is highly conserved among vertebrate classes. Fgf9 was expressed in the ovary of an adult frog strongly, but in the lung weakly. In contrast, the Fgf9 mRNA was hardly detected in the kidney, testis and muscle. Moreover, Fgf9 did not show a sexually dimorphic expression pattern during sex determination in R. rugosa. The results, taken together, suggest that both WT1 and Fgf9 are expressed in the indifferent gonad prior to sex determination without any difference in the expression between males and females. Thus, it seems unlikely that they are a key factor to initiate the divergence leading to testicular or ovarian differentiation in R. rugosa.",
    keywords = "cDNA, Fgf9, Gonad, Rana rugosa, RT-PCR, WT1",
    author = "Yasutaka Yamamura and Sho Aoyama and Yuki Oshima and Takuma Kato and Noboru Osawa and Masahisa Nakamura",
    year = "2005",
    month = "9",
    doi = "10.2108/zsj.22.1045",
    language = "English",
    volume = "22",
    pages = "1045--1050",
    journal = "Zoological Science",
    issn = "0289-0003",
    publisher = "Zoological Society of Japan",
    number = "9",

    }

    TY - JOUR

    T1 - Molecular cloning and expression in gonad of Rana rugosa WT1 and Fgf9

    AU - Yamamura, Yasutaka

    AU - Aoyama, Sho

    AU - Oshima, Yuki

    AU - Kato, Takuma

    AU - Osawa, Noboru

    AU - Nakamura, Masahisa

    PY - 2005/9

    Y1 - 2005/9

    N2 - Sry (sex-determining region on the Y chromosome) is required for testicular differentiation in mammals. In addition to Sry, other genes such as WT1, Fgf9, Dax1, Dmrt1 and Sox9 are widely accepted to be involved in the sex determination in vertebrates. However, the roles of these genes during sex determination still remain unclear in amphibians. This study was undertaken to examine the expression of WT1 and Fgf9 in the developing gonad of amphibians. We first isolated the WT1 cDNA from the frog Rana rugosa. Like WT1 in mice, R. rugosa WT1 showed 2 isoforms; i.e., one had an additional 3 amino acids, KTS, included between the third and fourth zinc fingers. However, 17 amino acids in exon 5 of mammalian WT1 could not be found in R. rugosa WT1, which is also the case in turtle and chicken. The mRNA of both isoforms (+KTS, -KTS) was detected in the lung, kidney and testis, but not in the ovary and muscle of adult frogs. The 2 isoforms were expressed first in the embryos at stage 23. Thereafter, the expressions remained constant in the gonad attached to mesonephros of both sexes during sex determination. We next isolated the R. rugosa Fgf9 cDNA encoding 208 amino acids. The amino acid sequence of Fgf9 had similarity greater than 92% with chicken, mouse and human Fgf9s, suggesting that Fgf9 is highly conserved among vertebrate classes. Fgf9 was expressed in the ovary of an adult frog strongly, but in the lung weakly. In contrast, the Fgf9 mRNA was hardly detected in the kidney, testis and muscle. Moreover, Fgf9 did not show a sexually dimorphic expression pattern during sex determination in R. rugosa. The results, taken together, suggest that both WT1 and Fgf9 are expressed in the indifferent gonad prior to sex determination without any difference in the expression between males and females. Thus, it seems unlikely that they are a key factor to initiate the divergence leading to testicular or ovarian differentiation in R. rugosa.

    AB - Sry (sex-determining region on the Y chromosome) is required for testicular differentiation in mammals. In addition to Sry, other genes such as WT1, Fgf9, Dax1, Dmrt1 and Sox9 are widely accepted to be involved in the sex determination in vertebrates. However, the roles of these genes during sex determination still remain unclear in amphibians. This study was undertaken to examine the expression of WT1 and Fgf9 in the developing gonad of amphibians. We first isolated the WT1 cDNA from the frog Rana rugosa. Like WT1 in mice, R. rugosa WT1 showed 2 isoforms; i.e., one had an additional 3 amino acids, KTS, included between the third and fourth zinc fingers. However, 17 amino acids in exon 5 of mammalian WT1 could not be found in R. rugosa WT1, which is also the case in turtle and chicken. The mRNA of both isoforms (+KTS, -KTS) was detected in the lung, kidney and testis, but not in the ovary and muscle of adult frogs. The 2 isoforms were expressed first in the embryos at stage 23. Thereafter, the expressions remained constant in the gonad attached to mesonephros of both sexes during sex determination. We next isolated the R. rugosa Fgf9 cDNA encoding 208 amino acids. The amino acid sequence of Fgf9 had similarity greater than 92% with chicken, mouse and human Fgf9s, suggesting that Fgf9 is highly conserved among vertebrate classes. Fgf9 was expressed in the ovary of an adult frog strongly, but in the lung weakly. In contrast, the Fgf9 mRNA was hardly detected in the kidney, testis and muscle. Moreover, Fgf9 did not show a sexually dimorphic expression pattern during sex determination in R. rugosa. The results, taken together, suggest that both WT1 and Fgf9 are expressed in the indifferent gonad prior to sex determination without any difference in the expression between males and females. Thus, it seems unlikely that they are a key factor to initiate the divergence leading to testicular or ovarian differentiation in R. rugosa.

    KW - cDNA

    KW - Fgf9

    KW - Gonad

    KW - Rana rugosa

    KW - RT-PCR

    KW - WT1

    UR - http://www.scopus.com/inward/record.url?scp=27644551423&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=27644551423&partnerID=8YFLogxK

    U2 - 10.2108/zsj.22.1045

    DO - 10.2108/zsj.22.1045

    M3 - Article

    VL - 22

    SP - 1045

    EP - 1050

    JO - Zoological Science

    JF - Zoological Science

    SN - 0289-0003

    IS - 9

    ER -