Nanos3 of the frog Rana rugosa: Molecular cloning and characterization

Maho Kodama, Madoka Yoshida, Masami Endo, Tohru Kobayashi, Akira Oike, Shigeki Yasumasu, Masahisa Nakamura

Research output: Contribution to journalArticlepeer-review

Abstract

Nanos is expressed in the primordial germ cells (PGCs) and also the germ cells of a variety of organisms as diverse as Drosophila, medaka fish, Xenopus and mouse. In Nanos3-deficient mice, PGCs fail to incorporate into the gonad and the size of the testis and ovary is thereby dramatically reduced. To elucidate the role of Nanos in an amphibian species, we cloned Nanos3 cDNA from the testis of the R. rugosa frog. RT-PCR analysis showed strong expression of Nanos3 mRNA in the testis of adult R. rugosa frogs, but expression was not sexually dimorphic during gonadal differentiation. In Nanos3-knockdown tadpoles produced by the CRISPR/Cas9 system, the number of germ cells decreased dramatically in the gonads of both male and female tadpoles before sex determination and thereafter. This was confirmed by three dimensional imaging of wild-type and Nanos3 knockdown gonads using serial sections immunostained for Vasa, a marker specific to germ cells. Taken together, these results suggest that Nanos3 protein function is conserved between R. rugosa and mouse.

Original languageEnglish
Pages (from-to)112-120
Number of pages9
JournalDevelopment Growth and Differentiation
Volume60
Issue number2
DOIs
Publication statusPublished - 2018 Feb 1
Externally publishedYes

Keywords

  • 3D imaging
  • CRISPR/Cas9
  • Nanos3
  • frog
  • germ cell
  • immunohistology

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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