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Production of Germ-Line Chimeras in Zebrafish

  • Taiju Saito
  • Rie Goto
  • Nicola Rivers
  • Etsuro Yamaha
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1920)

Abstract

The induction of germ-line chimerism in fish is a strategy for the reproduction of endangered or genetically valuable fish species. Chimeras can be created by transplanting a single primordial germ cell or multiple blastomeres from a donor into a sterile host embryo. When the host reaches sexual maturity, it will produce donor-originating gametes throughout its reproductive life span. This technique provides unique experimental conditions for basic biology research in model fish species like zebrafish. The success of cell transplantation relies on the effective sterilization of host embryos, the correct identification of developing germ cells, and the synchronization of migratory cues between the host and the transplanted cells. Developments in non-transgenic methods of germ cell ablation and identification have made germ cell transplantation more applicable to use in conservation and aquaculture. In this chapter, we provide a protocol for germ cell labeling by injection of chimeric RNA or FITC-dextran, the sterilization of host embryos using an antisense morpholino oligonucleotide, and two methods for producing germ-line chimeras in zebrafish: single primordial germ cell transplant and blastomere transplant.

Key words

Zebrafish Primordial germ cells Germ cell transplantation Blastomere transplant Host embryo sterilization 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Taiju Saito
    • 1
  • Rie Goto
    • 1
  • Nicola Rivers
    • 2
  • Etsuro Yamaha
    • 3
  1. 1.South Ehime Fisheries Research Center, Ehime UniversityEhimeJapan
  2. 2.Education Program in Reproduction and Development, Department of Obstetrics and GynaecologyMonash UniversityClaytonAustralia
  3. 3.Nanae Fresh Water LaboratoryField Science Center for Northern Biosphere, Hokkaido UniversityHokkaidoJapan

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