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Inducible Male Infertility by Targeted Cell Ablation in Zebrafish Testis

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Abstract

To generate a zebrafish model of inducible male sterility, we expressed an Escherichia coli nitroreductase (Ntr) gene in the male germ line of zebrafish. The Ntr gene encodes an enzyme that can convert prodrugs such as metronidazole (Met) to cytotoxins. A fusion protein eGFP:Ntr (fusing Ntr to eGFP) under control of ∼ 2 kb putative promoters of the zebrafish testis-specific genes, A-kinase anchoring protein-associated protein (Asp), outer dense fibers (Odf), and sperm acrosomal membrane-associated protein (Sam) was expressed in the male germ line. Three independent and four compound transgenic zebrafish lines expressing eGFP:Ntr were established. Female carriers were fertile, while males exhibited different levels of sterility and appeared normal, otherwise. Developmental analysis shows that germ cells survived and testes were normal before Met treatment, but that the testes of all male transgenic zebrafish exhibited variously depleted prospermatogonia after Met treatment. Particularly in a triple-transgenic line, Tg(AOS-eGFP:Ntr)[Tg(Asp-eGFP:Ntr; Odf-eGFP:Ntr; Sam-eGFP:Ntr)], the transgenic males had very small testes that were virtually devoid of germ cells, and the residual germ cells had almost completely disappeared after 2 weeks of Met treatment. These zebrafish transgenic lines show the complete testis specificity of inducible male sterility after Met treatment and reveal a period of the Ntr/Met ablation activity just prior to formation of the definitive adult spermatogonial cell population. This study demonstrates that combined genetic and pharmacological methods for developing an “infertile breeding technology” have practical application in controlling genetically modified (GM) fish breeding and meet the standards of biological and environment safety for other GM species.

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Acknowledgments

We thank Yin-Shan Chang and Yun-Sheng Shieh for assistance with zebrafish maintenance and breeding. This work was supported by grants from Taiwan National Science Council grant NSC 97-2317-B-019-001 and Ministry of Education for Promoting Academic Excellence of Center for Marine Bioscience Biotechnology (CMBB) grant 98529002AB to Guor Mour Her.

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Author notes

  1. Chia-Chun Hsu

    Present address: Department of Radiology, Buddhist Tzu Chi General Hospital, Taichung Branch, No. 66, Sec. 1, Fongsing Rd., Tanzih Township, Taichung County 427, Taichung, Taiwan, Republic of China

Authors and Affiliations

  1. Department of Radiology, Tri-Service General Hospital, 325, Cheng-Kung Road Sec. 2, Taipei, 114, Taiwan, Republic of China

    Chia-Chun Hsu

  2. Institute of Bioscience and Biotechnology, National Taiwan Ocean University (Room 202), 2 Pei Ning Road, Keelung, 20224, Taiwan, Republic of China

    Min-Fon Hou & Guor Mour Her

  3. Institute of Biotechnology, National Cheng-Kung University, Tainan, 701, Taiwan, Republic of China

    Jiann-Ruey Hong

  4. Institute of Cellular and Organismic Biology, Academia Sinica, 128, Academia Road, Sec. 2, Taipei, 115, Taiwan, Republic of China

    Jen-Leih Wu

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  1. Chia-Chun Hsu
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  2. Min-Fon Hou
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Correspondence to Guor Mour Her.

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Grant Sponsor

Taiwan National Science Council grant NSC 97-2317-B-019-001 and Ministry of Education for Promoting Academic Excellence of Center for Marine Bioscience Biotechnology (CMBB) grant 98529002AB

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Hsu, CC., Hou, MF., Hong, JR. et al. Inducible Male Infertility by Targeted Cell Ablation in Zebrafish Testis. Mar Biotechnol 12, 466–478 (2010). https://doi.org/10.1007/s10126-009-9248-4

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