Abstract
Many fish species exhibit remarkable sexual dimorphism, with males possessing numerous advantageous traits for commercial production by aquaculture such as faster growth rate, more efficient food energy utilization for muscle development, and better breeding performance. Several studies have shown that a decrease in the number of primordial germ cells (PGCs) during early development leads predominantly to male progeny. In this study, we developed a method to obtain all-male zebrafish (Danio rerio) by targeted PGC ablation using the nitroreductase/metronidazole (NTR/Mtz) system. Embryos generated by female heterozygous Tg(nanos3:nfsB-mCherry-nanos3 3′UTR) and male wild-types (WTs) were treated with vehicle or Mtz. Compared to vehicle-treated controls, 5.0 and 10.0 mM Mtz treatment for 24 h significantly reduced the number of PGCs and yielded an exclusively male phenotype in adulthood. The gonads of offspring treated with 5.0 mM Mtz exhibited relatively normal morphology and histological characteristics. Furthermore, these males were able to chase females, spawn, and produce viable offspring, while about 20.0% of males treated with 10.0 mM Mtz were unable to produce viable offspring. The 5.0 mM Mtz treatment protocol may thus be suitable for large-scale production of fertile male offspring. Moreover, about half of these males were WT as evidenced by the absence of nfsB gene expression. It may thus be possible to breed an all-male WT fish population by Mtz-mediated PGC ablation.
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This work was supported by the Guangdong Basic and Applied Basic Research Foundation (2021A1515010875) and the Weifang University Doctor Initiation Fund Project (208-44121015).
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Conceptualization, Fang Wang and Kai-Bin Li; data curation, Yong-Yong Feng; formal analysis, Xu-Guang Wang; funding acquisition, Jian Zhao and Kai-Bin Li; investigation, Fang Wang and Mi Ou; project administration, Kun-Ci Chen; visualization, Fang Wang and Xin-Cheng Zhang; writing-original draft, Fang Wang and Mi Ou; writing-review and editing, Kai-Bin Li. All authors have read and agreed to the published version of the manuscript.
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The Laboratory Animal Ethics Committee of Pearl River Fisheries Research Institute, CAFS under contract (LAEC-PRFRI-2021-01-02 ) approved this experiment, and the experimental process complied with international guidelines for the ethical use of animals in research.
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Wang, F., Feng, YY., Wang, XG. et al. Production of all-male non-transgenic zebrafish by conditional primordial germ cell ablation. Fish Physiol Biochem 49, 1215–1227 (2023). https://doi.org/10.1007/s10695-023-01252-y
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DOI: https://doi.org/10.1007/s10695-023-01252-y