Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation

Abstract 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 n...

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Autores principales:	Zoran Marinović, Qian Li, Jelena Lujić, Yoshiko Iwasaki, Zsolt Csenki, Béla Urbányi, Goro Yoshizaki, Ákos Horváth
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0a1824f86c6e441c9c6c5e03f09b4428
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spelling	oai:doaj.org-article:0a1824f86c6e441c9c6c5e03f09b44282021-12-02T15:08:45ZPreservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation10.1038/s41598-019-50169-12045-2322https://doaj.org/article/0a1824f86c6e441c9c6c5e03f09b44282019-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-50169-1https://doaj.org/toc/2045-2322Abstract 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.Zoran MarinovićQian LiJelena LujićYoshiko IwasakiZsolt CsenkiBéla UrbányiGoro YoshizakiÁkos HorváthNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-10 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zoran Marinović Qian Li Jelena Lujić Yoshiko Iwasaki Zsolt Csenki Béla Urbányi Goro Yoshizaki Ákos Horváth Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
description	Abstract 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.
format	article
author	Zoran Marinović Qian Li Jelena Lujić Yoshiko Iwasaki Zsolt Csenki Béla Urbányi Goro Yoshizaki Ákos Horváth
author_facet	Zoran Marinović Qian Li Jelena Lujić Yoshiko Iwasaki Zsolt Csenki Béla Urbányi Goro Yoshizaki Ákos Horváth
author_sort	Zoran Marinović
title	Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
title_short	Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
title_full	Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
title_fullStr	Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
title_full_unstemmed	Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
title_sort	preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/0a1824f86c6e441c9c6c5e03f09b4428
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Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation

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