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Identification of Germ-Line Stem Cells in Zebrafish

  • Bruce W. DraperEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1463)

Abstract

Both male and female zebrafish have a population of germ-line stem cells that produce gametes throughout the life of the fish. These cells localize to specific regions in the gonads and can be identified because they uniquely express the nanos2 gene, which encodes a conserved regulator of translation. A method is presented here for identifying germ-line stem cells in the ovary and testis using a combined protocol for whole-mount fluorescent RNA in situ hybridization to detect nanos2 mRNA and immunofluorescence to detect the pan-germ cell marker Vasa.

Key words

Zebrafish germ-line stem cells Oogonial stem cell Spermatogonial stem cell Zebrafish ovary Zebrafish testis nanos2 Vasa Fluorescent RNA in situ hybridization Confocal immunofluorescence 

Notes

Acknowledgments

The author would like to thank Holger Knaut for the anti-Vasa antibodies. This work was supported by a NIH grant R01-HD081551.

References

  1. 1.
    Draper BW, McCallum CM, Moens CB (2007) nanos1 is required to maintain oocyte production in adult zebrafish. Dev Biol 305:589–598CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Beer RL, Draper BW (2013) nanos3 maintains germline stem cells and expression of the conserved germline stem cell gene nanos2 in the zebrafish ovary. Dev Biol 374:308–318CrossRefPubMedGoogle Scholar
  3. 3.
    Sada A, Suzuki A, Suzuki H, Saga Y (2009) The RNA-binding protein nanos2 is required to maintain murine spermatogonial stem cells. Science 325:1394–1398CrossRefPubMedGoogle Scholar
  4. 4.
    Nakamura S, Kobayashi K, Nishimura T et al (2010) Identification of germline stem cells in the ovary of the teleost medaka. Science 328:1561–1563CrossRefPubMedGoogle Scholar
  5. 5.
    Thisse B, Thisse C (2014) In situ hybridization on whole-mount zebrafish embryos and young larvae. In: Nielsen BS (ed) In situ hybridization protocols, vol 1211, Methods in molecular biology. Springer Science + Business Media, New York, NY, pp 53–67Google Scholar
  6. 6.
    Draper BW (2012) Identification of oocyte progenitor cells in the zebrafish ovary. In: Mace KA, Braun KM (eds) Progenitor cells: methods and protocols, vol 916, Methods in molecular biology. Springer Science + Business Media, New York, NY, pp 157–165CrossRefGoogle Scholar
  7. 7.
    Knaut H, Pelegri F, Bohmann K et al (2000) Zebrafish vasa RNA but not its protein is a component of the germ plasm and segregates asymmetrically before germline specification. J Cell Biol 149:875–888CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Molecular and Cellular BiologyUniversity of CaliforniaDavisUSA

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