학술논문
An inducible model of chronic hyperglycemia.
Document Type
Academic Journal
Author
Tucker TR; Department of Developmental and Cell Biology, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Knitter CA; Department of Molecular Biology and Biochemistry, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Khoury DM; Department of Developmental and Cell Biology, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Eshghi S; Department of Developmental and Cell Biology, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Tran S; Department of Developmental and Cell Biology, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Sharrock AV; School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand.; Wiles TJ; Department of Molecular Biology and Biochemistry, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.; Ackerley DF; School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand.; Mumm JS; Department of Ophthalmology, The Wilmer Eye Institute, Johns Hopkins University, Baltimore, MD, USA.; Parsons MJ; Department of Developmental and Cell Biology, University of California, Irvine, Natural Sciences II, Irvine, CA 92697, USA.
Source
Publisher: Company of Biologists Ltd Country of Publication: England NLM ID: 101483332 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1754-8411 (Electronic) Linking ISSN: 17548403 NLM ISO Abbreviation: Dis Model Mech Subsets: MEDLINE
Subject
Language
English
Abstract
Transgene driven expression of Escherichia coli nitroreductase (NTR1.0) renders animal cells susceptible to the antibiotic metronidazole (MTZ). Many NTR1.0/MTZ ablation tools have been reported in zebrafish, which have significantly impacted regeneration studies. However, NTR1.0-based tools are not appropriate for modeling chronic cell loss as prolonged application of the required MTZ dose (10 mM) is deleterious to zebrafish health. We established that this dose corresponds to the median lethal dose (LD50) of MTZ in larval and adult zebrafish and that it induced intestinal pathology. NTR2.0 is a more active nitroreductase engineered from Vibrio vulnificus NfsB that requires substantially less MTZ to induce cell ablation. Here, we report on the generation of two new NTR2.0-based zebrafish lines in which acute β-cell ablation can be achieved without MTZ-associated intestinal pathology. For the first time, we were able to sustain β-cell loss and maintain elevated glucose levels (chronic hyperglycemia) in larvae and adults. Adult fish showed significant weight loss, consistent with the induction of a diabetic state, indicating that this paradigm will allow the modeling of diabetes and associated pathologies.
Competing Interests: Competing interests J.S.M. has been awarded patents for the creation (US patent no. 7,514,595) and use of zebrafish expressing NTR enzymes for gene (US patent no. 8,071,838) and drug discovery (US patent no. 8,431,768) applications. J.S.M. serves as a consultant at Luminomics, a biotechnology start-up that offers phenotypic drug-based screening services. The remaining authors declare no competing interests.
(© 2023. Published by The Company of Biologists Ltd.)
Competing Interests: Competing interests J.S.M. has been awarded patents for the creation (US patent no. 7,514,595) and use of zebrafish expressing NTR enzymes for gene (US patent no. 8,071,838) and drug discovery (US patent no. 8,431,768) applications. J.S.M. serves as a consultant at Luminomics, a biotechnology start-up that offers phenotypic drug-based screening services. The remaining authors declare no competing interests.
(© 2023. Published by The Company of Biologists Ltd.)