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An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention

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 re...

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Autores principales: Kulkarni, Abhishek A., Conteh, Abass M., Sorrell, Cody A., Mirmira, Anjali, Tersey, Sarah A., Mirmira, Raghavendra G., Linnemann, Amelia K., Anderson, Ryan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5896207/
https://www.ncbi.nlm.nih.gov/pubmed/29785241
http://dx.doi.org/10.1155/2018/1324739
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author Kulkarni, Abhishek A.
Conteh, Abass M.
Sorrell, Cody A.
Mirmira, Anjali
Tersey, Sarah A.
Mirmira, Raghavendra G.
Linnemann, Amelia K.
Anderson, Ryan M.
author_facet Kulkarni, Abhishek A.
Conteh, Abass M.
Sorrell, Cody A.
Mirmira, Anjali
Tersey, Sarah A.
Mirmira, Raghavendra G.
Linnemann, Amelia K.
Anderson, Ryan M.
author_sort Kulkarni, Abhishek A.
collection PubMed
description 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.
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spelling pubmed-58962072018-05-21 An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention Kulkarni, Abhishek A. Conteh, Abass M. Sorrell, Cody A. Mirmira, Anjali Tersey, Sarah A. Mirmira, Raghavendra G. Linnemann, Amelia K. Anderson, Ryan M. Oxid Med Cell Longev Research Article 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. Hindawi 2018-03-28 /pmc/articles/PMC5896207/ /pubmed/29785241 http://dx.doi.org/10.1155/2018/1324739 Text en Copyright © 2018 Abhishek A. Kulkarni et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Kulkarni, Abhishek A.
Conteh, Abass M.
Sorrell, Cody A.
Mirmira, Anjali
Tersey, Sarah A.
Mirmira, Raghavendra G.
Linnemann, Amelia K.
Anderson, Ryan M.
An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title_full An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title_fullStr An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title_full_unstemmed An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title_short An In Vivo Zebrafish Model for Interrogating ROS-Mediated Pancreatic β-Cell Injury, Response, and Prevention
title_sort in vivo zebrafish model for interrogating ros-mediated pancreatic β-cell injury, response, and prevention
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5896207/
https://www.ncbi.nlm.nih.gov/pubmed/29785241
http://dx.doi.org/10.1155/2018/1324739
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