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Regeneration of the pancreas in adult zebrafish.

Moss JB, Koustubhan P, Greenman M, Parsons MJ, Walter I, Moss LG - Diabetes (2009)

Bottom Line: Dividing cells were primarily associated with affected islets and ducts but, with the exception of surgical partial pancreatectomy, were not extensively beta-cells.The ability of the zebrafish to regenerate a functional pancreas using chemical, genetic, and surgical approaches enabled us to identify patterns of cell proliferation in islets and ducts.Further study of the origin and contribution of proliferating cells in reestablishing islet function could provide strategies for treating human diseases.

View Article: PubMed Central - PubMed

Affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina, USA. jennifer.b.moss@duke.edu

ABSTRACT

Objective: Regenerating organs in diverse biological systems have provided clues to processes that can be harnessed to repair damaged tissue. Adult mammalian beta-cells have a limited capacity to regenerate, resulting in diabetes and lifelong reliance on insulin. Zebrafish have been used as a model for the regeneration of many organs. We demonstrate the regeneration of adult zebrafish pancreatic beta-cells. This nonmammalian model can be used to define pathways for islet-cell regeneration in humans.

Research design and methods: Adult transgenic zebrafish were injected with a single high dose of streptozotocin or metronidazole and anesthetized at 3, 7, or 14 days or pancreatectomized. Blood glucose measurements were determined and gut sections were analyzed using specific endocrine, exocrine, and duct cell markers as well as markers for dividing cells.

Results: Zebrafish recovered rapidly without the need for insulin injections, and normoglycemia was attained within 2 weeks. Although few proliferating cells were present in vehicles, ablation caused islet destruction and a striking increase of proliferating cells, some of which were Pdx1 positive. Dividing cells were primarily associated with affected islets and ducts but, with the exception of surgical partial pancreatectomy, were not extensively beta-cells.

Conclusions: The ability of the zebrafish to regenerate a functional pancreas using chemical, genetic, and surgical approaches enabled us to identify patterns of cell proliferation in islets and ducts. Further study of the origin and contribution of proliferating cells in reestablishing islet function could provide strategies for treating human diseases.

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Related in: MedlinePlus

β-cell destruction by STZ. A: GFP fluorescence of zebrafish pancreatic explants after 3 days in culture (100×). B: Intact zebrafish: lateral views of right side without skin (50×). Top panel: GFP fluorescence in the main pancreas (rostral) and auxiliary islets (green) of vehicle-injected zebrafish. Bottom panel: STZ-injected zebrafish killed after 3 days. C: Vehicle-injected, killed InsGFP zebrafish. Main pancreas was imaged after in situ treatment with propidium iodide (propidium iodide: red fluorescence). D: STZ-injected zebrafish pancreas + propidium iodide after 3 days. (A high-quality digital representation of this figure is available in the online issue.)
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Figure 1: β-cell destruction by STZ. A: GFP fluorescence of zebrafish pancreatic explants after 3 days in culture (100×). B: Intact zebrafish: lateral views of right side without skin (50×). Top panel: GFP fluorescence in the main pancreas (rostral) and auxiliary islets (green) of vehicle-injected zebrafish. Bottom panel: STZ-injected zebrafish killed after 3 days. C: Vehicle-injected, killed InsGFP zebrafish. Main pancreas was imaged after in situ treatment with propidium iodide (propidium iodide: red fluorescence). D: STZ-injected zebrafish pancreas + propidium iodide after 3 days. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: We initially tested whether zebrafish islets were susceptible to the toxic effects of STZ, a nitrosourea causing DNA damage after entering β-cells through the Glut2 receptor. Adult zebrafish containing a stable transgene expressing GFP from the zebrafish insulin promoter only in β-cells (InsGFP [(16)]) were anesthetized, and the main GFP-positive pancreatic tissue was excised and incubated for 3 days in media used for culturing rat primary islets (17). At 0 or 20 mmol/l STZ, no gross changes in GFP fluorescence or morphology were detectable. However, at 100 and 200 mmol/l STZ, significant loss of GFP-positive cells was observed in culture (Fig. 1A). These data provided the basis for examining the physiological effects of β-cell destruction in living InsGFP zebrafish.


Regeneration of the pancreas in adult zebrafish.

Moss JB, Koustubhan P, Greenman M, Parsons MJ, Walter I, Moss LG - Diabetes (2009)

β-cell destruction by STZ. A: GFP fluorescence of zebrafish pancreatic explants after 3 days in culture (100×). B: Intact zebrafish: lateral views of right side without skin (50×). Top panel: GFP fluorescence in the main pancreas (rostral) and auxiliary islets (green) of vehicle-injected zebrafish. Bottom panel: STZ-injected zebrafish killed after 3 days. C: Vehicle-injected, killed InsGFP zebrafish. Main pancreas was imaged after in situ treatment with propidium iodide (propidium iodide: red fluorescence). D: STZ-injected zebrafish pancreas + propidium iodide after 3 days. (A high-quality digital representation of this figure is available in the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2712797&req=5

Figure 1: β-cell destruction by STZ. A: GFP fluorescence of zebrafish pancreatic explants after 3 days in culture (100×). B: Intact zebrafish: lateral views of right side without skin (50×). Top panel: GFP fluorescence in the main pancreas (rostral) and auxiliary islets (green) of vehicle-injected zebrafish. Bottom panel: STZ-injected zebrafish killed after 3 days. C: Vehicle-injected, killed InsGFP zebrafish. Main pancreas was imaged after in situ treatment with propidium iodide (propidium iodide: red fluorescence). D: STZ-injected zebrafish pancreas + propidium iodide after 3 days. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: We initially tested whether zebrafish islets were susceptible to the toxic effects of STZ, a nitrosourea causing DNA damage after entering β-cells through the Glut2 receptor. Adult zebrafish containing a stable transgene expressing GFP from the zebrafish insulin promoter only in β-cells (InsGFP [(16)]) were anesthetized, and the main GFP-positive pancreatic tissue was excised and incubated for 3 days in media used for culturing rat primary islets (17). At 0 or 20 mmol/l STZ, no gross changes in GFP fluorescence or morphology were detectable. However, at 100 and 200 mmol/l STZ, significant loss of GFP-positive cells was observed in culture (Fig. 1A). These data provided the basis for examining the physiological effects of β-cell destruction in living InsGFP zebrafish.

Bottom Line: Dividing cells were primarily associated with affected islets and ducts but, with the exception of surgical partial pancreatectomy, were not extensively beta-cells.The ability of the zebrafish to regenerate a functional pancreas using chemical, genetic, and surgical approaches enabled us to identify patterns of cell proliferation in islets and ducts.Further study of the origin and contribution of proliferating cells in reestablishing islet function could provide strategies for treating human diseases.

View Article: PubMed Central - PubMed

Affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina, USA. jennifer.b.moss@duke.edu

ABSTRACT

Objective: Regenerating organs in diverse biological systems have provided clues to processes that can be harnessed to repair damaged tissue. Adult mammalian beta-cells have a limited capacity to regenerate, resulting in diabetes and lifelong reliance on insulin. Zebrafish have been used as a model for the regeneration of many organs. We demonstrate the regeneration of adult zebrafish pancreatic beta-cells. This nonmammalian model can be used to define pathways for islet-cell regeneration in humans.

Research design and methods: Adult transgenic zebrafish were injected with a single high dose of streptozotocin or metronidazole and anesthetized at 3, 7, or 14 days or pancreatectomized. Blood glucose measurements were determined and gut sections were analyzed using specific endocrine, exocrine, and duct cell markers as well as markers for dividing cells.

Results: Zebrafish recovered rapidly without the need for insulin injections, and normoglycemia was attained within 2 weeks. Although few proliferating cells were present in vehicles, ablation caused islet destruction and a striking increase of proliferating cells, some of which were Pdx1 positive. Dividing cells were primarily associated with affected islets and ducts but, with the exception of surgical partial pancreatectomy, were not extensively beta-cells.

Conclusions: The ability of the zebrafish to regenerate a functional pancreas using chemical, genetic, and surgical approaches enabled us to identify patterns of cell proliferation in islets and ducts. Further study of the origin and contribution of proliferating cells in reestablishing islet function could provide strategies for treating human diseases.

Show MeSH
Related in: MedlinePlus