An in vivo zebrafish model for interrogating ROS-mediated pancreatic β-cell injury, response, and prevention

Abhishek A. Kulkarni, Abass M. Conteh, Cody A. Sorrell, Anjali Mirmira, Sarah A. Tersey, Raghavendra G. Mirmira, Amelia K. Linnemann, Ryan M. Anderson

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

It is well known that a chronic state of elevated reactive oxygen species (ROS) in pancreatic β-cells impairs their ability to release insulin in response to elevated plasma glucose. Moreover, at its extreme, unmitigated ROS drives regulated cell death. This dysfunctional state of ROS buildup can result both from genetic predisposition and environmental factors such as obesity and overnutrition. Importantly, excessive ROS buildup may underlie metabolic pathologies such as type 2 diabetes mellitus. The ability to monitor ROS dynamics in β-cells in situ and to manipulate it via genetic, pharmacological, and environmental means would accelerate the development of novel therapeutics that could abate this pathology. Currently, there is a lack of models with these attributes that are available to the field. In this study, we use a zebrafish model to demonstrate that ROS can be generated in a β-cell-specific manner using a hybrid chemical genetic approach. Using a transgenic nitroreductase-expressing zebrafish line, Tg(ins:Flag-NTR)s950, treated with the prodrug metronidazole (MTZ), we found that ROS is rapidly and explicitly generated in β-cells. Furthermore, the level of ROS generated was proportional to the dosage of prodrug added to the system. At high doses of MTZ, caspase 3 was rapidly cleaved, β-cells underwent regulated cell death, and macrophages were recruited to the islet to phagocytose the debris. Based on our findings, we propose a model for the mechanism of NTR/MTZ action in transgenic eukaryotic cells and demonstrate the robust utility of this system to model ROS-related disease pathology.

Original languageEnglish (US)
Article number1324739
JournalOxidative medicine and cellular longevity
Volume2018
DOIs
StatePublished - 2018

ASJC Scopus subject areas

  • Biochemistry
  • Aging
  • Cell Biology

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