Inducible male infertility by targeted cell ablation in zebrafish testis.

Marine Biotechnology
Chia-Chun HsuGuor Mour Her

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 approximately 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 disapp...Continue Reading

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