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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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Regeneration after pancreatectomy. A and B: Right side of intact, living zebrafish before (sham, A) and 14 days after (Ptx, B) surgical removal of the GFP+ pancreas (red outline). The tip of the forceps used to remove the pancreas is visible (100× magnification). C: Paraffin section of sham-operated pancreas with few PCNA+ dividing cells (PCNA, red) except in the intestine (Int). β-Cells are green (arrow; 200× magnification). D: Many red PCNA+ dividing cells in ducts (arrowhead) and in nuclei of regenerating β-cells (yellow; 200× magnification). (A high-quality digital representation of this figure is available on the online issue.)
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Figure 5: Regeneration after pancreatectomy. A and B: Right side of intact, living zebrafish before (sham, A) and 14 days after (Ptx, B) surgical removal of the GFP+ pancreas (red outline). The tip of the forceps used to remove the pancreas is visible (100× magnification). C: Paraffin section of sham-operated pancreas with few PCNA+ dividing cells (PCNA, red) except in the intestine (Int). β-Cells are green (arrow; 200× magnification). D: Many red PCNA+ dividing cells in ducts (arrowhead) and in nuclei of regenerating β-cells (yellow; 200× magnification). (A high-quality digital representation of this figure is available on the online issue.)

Mentions: Anesthetized zebrafish were pancreatectomized (Ptx) or manipulated with forceps without removing GFP-positive tissue (sham-operated control). We removed as much of the GFP-positive tissue as possible without severely damaging the liver, gall bladder, or spleen. After 7 days, the wound had healed and GFP-positive cells were photographed in situ (Fig. 5A vs. B). Blood glucose readings in Ptx fish killed after 14 days were on average 78.9 mg/dl (n = 6). Sham-operated animals had normal blood sugars (52.4 mg/dl, n = 4). Fibrotic tissue was observed along with a proliferation of small islets. A histological assessment indicated that, like the STZ-treated fish, ductal elements were associated with regenerating tissue. Dividing cells were not prominent in the pancreas of sham-operated animals, whereas numerous PCNA-positive cells were found in pancreatic ducts (Fig. 5C). Surprisingly, unlike the chemical-ablation models, PCNA co-stained many insulin-positive cells (Fig. 5D). The partial physical ablation caused by pancreatectomy resulted in an increased division of existing β-cells that was not observed after chemical ablation. In contrast, ductal hyperplasia and a prominent association of regenerating islets with ducts were required for both wound repair and the regeneration we observed after STZ or MET treatment.


Regeneration of the pancreas in adult zebrafish.

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

Regeneration after pancreatectomy. A and B: Right side of intact, living zebrafish before (sham, A) and 14 days after (Ptx, B) surgical removal of the GFP+ pancreas (red outline). The tip of the forceps used to remove the pancreas is visible (100× magnification). C: Paraffin section of sham-operated pancreas with few PCNA+ dividing cells (PCNA, red) except in the intestine (Int). β-Cells are green (arrow; 200× magnification). D: Many red PCNA+ dividing cells in ducts (arrowhead) and in nuclei of regenerating β-cells (yellow; 200× magnification). (A high-quality digital representation of this figure is available on the online issue.)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 5: Regeneration after pancreatectomy. A and B: Right side of intact, living zebrafish before (sham, A) and 14 days after (Ptx, B) surgical removal of the GFP+ pancreas (red outline). The tip of the forceps used to remove the pancreas is visible (100× magnification). C: Paraffin section of sham-operated pancreas with few PCNA+ dividing cells (PCNA, red) except in the intestine (Int). β-Cells are green (arrow; 200× magnification). D: Many red PCNA+ dividing cells in ducts (arrowhead) and in nuclei of regenerating β-cells (yellow; 200× magnification). (A high-quality digital representation of this figure is available on the online issue.)
Mentions: Anesthetized zebrafish were pancreatectomized (Ptx) or manipulated with forceps without removing GFP-positive tissue (sham-operated control). We removed as much of the GFP-positive tissue as possible without severely damaging the liver, gall bladder, or spleen. After 7 days, the wound had healed and GFP-positive cells were photographed in situ (Fig. 5A vs. B). Blood glucose readings in Ptx fish killed after 14 days were on average 78.9 mg/dl (n = 6). Sham-operated animals had normal blood sugars (52.4 mg/dl, n = 4). Fibrotic tissue was observed along with a proliferation of small islets. A histological assessment indicated that, like the STZ-treated fish, ductal elements were associated with regenerating tissue. Dividing cells were not prominent in the pancreas of sham-operated animals, whereas numerous PCNA-positive cells were found in pancreatic ducts (Fig. 5C). Surprisingly, unlike the chemical-ablation models, PCNA co-stained many insulin-positive cells (Fig. 5D). The partial physical ablation caused by pancreatectomy resulted in an increased division of existing β-cells that was not observed after chemical ablation. In contrast, ductal hyperplasia and a prominent association of regenerating islets with ducts were required for both wound repair and the regeneration we observed after STZ or MET treatment.

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