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Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation.
Marinovic, Zoran; Li, Qian; Lujic, Jelena; Iwasaki, Yoshiko; Csenki, Zsolt; Urbányi, Béla; Yoshizaki, Goro; Horváth, Ákos.
Afiliación
  • Marinovic Z; Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100, Gödöllo, Hungary.
  • Li Q; Department of Marine Biosciences, Tokyo University of Marine Science and Technology, 108-8477, Tokyo, Japan.
  • Lujic J; Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100, Gödöllo, Hungary. lujicjelena@gmail.com.
  • Iwasaki Y; Department of Marine Biosciences, Tokyo University of Marine Science and Technology, 108-8477, Tokyo, Japan.
  • Csenki Z; Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100, Gödöllo, Hungary.
  • Urbányi B; Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100, Gödöllo, Hungary.
  • Yoshizaki G; Department of Marine Biosciences, Tokyo University of Marine Science and Technology, 108-8477, Tokyo, Japan.
  • Horváth Á; Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100, Gödöllo, Hungary.
Sci Rep ; 9(1): 13861, 2019 09 25.
Article en En | MEDLINE | ID: mdl-31554831
Zebrafish is one of the most commonly used model organisms in biomedical, developmental and genetic research. The production of several thousands of transgenic lines is leading to difficulties in maintaining valuable genetic resources as cryopreservation protocols for eggs and embryos are not yet developed. In this study, we utilized testis cryopreservation (through both slow-rate freezing and vitrification) and spermatogonia transplantation as effective methods for long-term storage and line reconstitution in zebrafish. During freezing, utilization of 1.3 M of dimethyl sulfoxide (Me2SO) displayed the highest spermatogonia viability (~60%), while sugar and protein supplementation had no effects. Needle-immersed vitrification also yielded high spermatogonia viability rates (~50%). Both optimal slow-rate freezing and vitrification protocols proved to be reproducible in six tested zebrafish lines after displaying viability rates of >50% in all lines. Both fresh and cryopreserved spermatogonia retained their ability to colonize the recipient gonads after intraperitoneal transplantation of vasa::egfp and actb:yfp spermatogonia into wild-type AB recipient larvae. Colonization rate was significantly higher in dnd-morpholino sterilized recipients than in non-sterilized recipients. Lastly, wild-type recipients produced donor-derived sperm and donor-derived offspring through natural spawning. The method demonstrated in this study can be used for long-term storage of valuable zebrafish genetic resources and for reconstitution of whole zebrafish lines which will greatly improve the current preservation practices.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espermatogonias / Testículo / Pez Cebra / Criopreservación Tipo de estudio: Guideline Límite: Animals Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espermatogonias / Testículo / Pez Cebra / Criopreservación Tipo de estudio: Guideline Límite: Animals Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Reino Unido