例如:"lncRNA", "apoptosis", "WRKY"

Nitroreductase-mediated gonadal dysgenesis for infertility control of genetically modified zebrafish.

Mar. Biotechnol.2010 Oct;12(5):569-78. doi:10.1007/s10126-009-9244-8. Epub 2009 Nov 27
Shao-Yang Hu 1 , Pei-Yu Lin , Chia-Hsuan Liao , Hong-Yi Gong , Gen-Hwa Lin , Koichi Kawakami , Jen-Leih Wu
Shao-Yang Hu 1 , Pei-Yu Lin , Chia-Hsuan Liao , Hong-Yi Gong , Gen-Hwa Lin , Koichi Kawakami , Jen-Leih Wu
+ et al

[No authors listed]

Author information
  • 1 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan.

摘要


Genetically modified (GM) fish with desirable features such as rapid growth, disease resistance, and cold tolerance, among other traits, have been established in aquaculture. However, commercially available GM fish are restricted because of global concerns over the incomplete assessments of food safety and ecological impact. The ecological impact concerns include gene flow and escape of the GM fish, which may cause extinction of wild natural fish stocks. Infertility control is a core technology for overcoming this obstacle. Although polyploidy technology, GnRH-specific antisense RNA, and against GnRH gene expression have been used to cause infertility in fish, these approaches are not 100% reliable and are not heritable. In the present study, zebrafish was used as a model to establish an inducible platform of infertility control in GM fish. Nitroreductase, which converts metronidazole substrate into cytotoxin, was fused with EGFP and expressed specifically by oocytes in the Tg(ZP:NTR-EGFP) by a zona pellucida promoter. Through consecutive immersion of metronidazole from 28 to 42 days posthatching, oocyte-specific EGFP expression was eliminated, and atrophy of the gonads was detected by anatomical analysis. These findings reveal that oocyte-specific nitroreductase-mediated catalysis of metronidazole blocks oogenesis and leads to an undeveloped oocyte. Furthermore, oocyte cell death via apoptosis was detected by a TUNEL assay. We found that the gonadal dysgenesis induced by metronidazole resulted in activation of the ovarian killer gene bok, which is a proapoptotic gene member of the Bcl-2 family and led to infertility. These results show that oocyte-specific nitroreductase-mediated catalysis of metronidazole can cause reliable infertility in zebrafish and could potentially be used as a model for other aquaculture fish species.