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Centroacinar Cells Are Progenitors That Contribute to Endocrine Pancreas Regeneration

View Article: PubMed Central - PubMed

ABSTRACT

Diabetes is associated with a paucity of insulin-producing β-cells. With the goal of finding therapeutic routes to treat diabetes, we aim to find molecular and cellular mechanisms involved in β-cell neogenesis and regeneration. To facilitate discovery of such mechanisms, we use a vertebrate organism where pancreatic cells readily regenerate. The larval zebrafish pancreas contains Notch-responsive progenitors that during development give rise to adult ductal, endocrine, and centroacinar cells (CACs). Adult CACs are also Notch responsive and are morphologically similar to their larval predecessors. To test our hypothesis that adult CACs are also progenitors, we took two complementary approaches: 1) We established the transcriptome for adult CACs. Using gene ontology, transgenic lines, and in situ hybridization, we found that the CAC transcriptome is enriched for progenitor markers. 2) Using lineage tracing, we demonstrated that CACs do form new endocrine cells after β-cell ablation or partial pancreatectomy. We concluded that CACs and their larval predecessors are the same cell type and represent an opportune model to study both β-cell neogenesis and β-cell regeneration. Furthermore, we show that in cftr loss-of-function mutants, there is a deficiency of larval CACs, providing a possible explanation for pancreatic complications associated with cystic fibrosis.

No MeSH data available.


CACs are progenitors of endocrine and ductal cell types. A–C: 4OHT injection labels 75% of Notch-responsive cells (green) with nuc-mCherry (red nuclei) in LT; Tp1:GFP fish. D–F: LT; ins:NTR regenerating islets at MTZ +7 days. nuc-mCherry labels insulin-expressing CAC progeny (yellow arrowheads). G–I: LT; Ptf1a:gfp fish at PPx +7 days. nuc-mCherry (red nuclei) labels regenerating duct, CACs (yellow asterisks), and insulin-expressing islet cells (white) but not eGFP-positive acinar cells (green). Anti-insulin (white), anti-glucagon (green in B), anti-mCherry (red), DAPI (blue). Scale bars = 25 μm.
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Figure 6: CACs are progenitors of endocrine and ductal cell types. A–C: 4OHT injection labels 75% of Notch-responsive cells (green) with nuc-mCherry (red nuclei) in LT; Tp1:GFP fish. D–F: LT; ins:NTR regenerating islets at MTZ +7 days. nuc-mCherry labels insulin-expressing CAC progeny (yellow arrowheads). G–I: LT; Ptf1a:gfp fish at PPx +7 days. nuc-mCherry (red nuclei) labels regenerating duct, CACs (yellow asterisks), and insulin-expressing islet cells (white) but not eGFP-positive acinar cells (green). Anti-insulin (white), anti-glucagon (green in B), anti-mCherry (red), DAPI (blue). Scale bars = 25 μm.

Mentions: To directly test whether CACs are a bona fide adult progenitor population, we used our cre-based strategy to lineage trace Notch-responsive cells (12,41). This system uses two transgenes: 1) a Notch-responsive cre driver, Tp1:CreERT2, and 2) the cre responder, βactin:lox-stop-lox-hmgb1-mCherry. Zebrafish carrying both transgenes are called lineage-tracing fish (LT). Addition of 4OHT to LT fish indelibly labels Notch-responsive cells with nuclear-mCherry (nuc-mCherry). To quantify the efficiency of 4OHT-dependent labeling in adults, we used LT fish that also carried the Notch-responsive reporter, Tp1:GFP. After injection of adult LT; Tp1:GFP fish with 4OHT daily for 3 days, 75% of GFP-positive cells were labeled with nuc-mCherry (n = 5) (Fig. 6A–C).


Centroacinar Cells Are Progenitors That Contribute to Endocrine Pancreas Regeneration
CACs are progenitors of endocrine and ductal cell types. A–C: 4OHT injection labels 75% of Notch-responsive cells (green) with nuc-mCherry (red nuclei) in LT; Tp1:GFP fish. D–F: LT; ins:NTR regenerating islets at MTZ +7 days. nuc-mCherry labels insulin-expressing CAC progeny (yellow arrowheads). G–I: LT; Ptf1a:gfp fish at PPx +7 days. nuc-mCherry (red nuclei) labels regenerating duct, CACs (yellow asterisks), and insulin-expressing islet cells (white) but not eGFP-positive acinar cells (green). Anti-insulin (white), anti-glucagon (green in B), anti-mCherry (red), DAPI (blue). Scale bars = 25 μm.
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Related In: Results  -  Collection

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Figure 6: CACs are progenitors of endocrine and ductal cell types. A–C: 4OHT injection labels 75% of Notch-responsive cells (green) with nuc-mCherry (red nuclei) in LT; Tp1:GFP fish. D–F: LT; ins:NTR regenerating islets at MTZ +7 days. nuc-mCherry labels insulin-expressing CAC progeny (yellow arrowheads). G–I: LT; Ptf1a:gfp fish at PPx +7 days. nuc-mCherry (red nuclei) labels regenerating duct, CACs (yellow asterisks), and insulin-expressing islet cells (white) but not eGFP-positive acinar cells (green). Anti-insulin (white), anti-glucagon (green in B), anti-mCherry (red), DAPI (blue). Scale bars = 25 μm.
Mentions: To directly test whether CACs are a bona fide adult progenitor population, we used our cre-based strategy to lineage trace Notch-responsive cells (12,41). This system uses two transgenes: 1) a Notch-responsive cre driver, Tp1:CreERT2, and 2) the cre responder, βactin:lox-stop-lox-hmgb1-mCherry. Zebrafish carrying both transgenes are called lineage-tracing fish (LT). Addition of 4OHT to LT fish indelibly labels Notch-responsive cells with nuclear-mCherry (nuc-mCherry). To quantify the efficiency of 4OHT-dependent labeling in adults, we used LT fish that also carried the Notch-responsive reporter, Tp1:GFP. After injection of adult LT; Tp1:GFP fish with 4OHT daily for 3 days, 75% of GFP-positive cells were labeled with nuc-mCherry (n = 5) (Fig. 6A–C).

View Article: PubMed Central - PubMed

ABSTRACT

Diabetes is associated with a paucity of insulin-producing β-cells. With the goal of finding therapeutic routes to treat diabetes, we aim to find molecular and cellular mechanisms involved in β-cell neogenesis and regeneration. To facilitate discovery of such mechanisms, we use a vertebrate organism where pancreatic cells readily regenerate. The larval zebrafish pancreas contains Notch-responsive progenitors that during development give rise to adult ductal, endocrine, and centroacinar cells (CACs). Adult CACs are also Notch responsive and are morphologically similar to their larval predecessors. To test our hypothesis that adult CACs are also progenitors, we took two complementary approaches: 1) We established the transcriptome for adult CACs. Using gene ontology, transgenic lines, and in situ hybridization, we found that the CAC transcriptome is enriched for progenitor markers. 2) Using lineage tracing, we demonstrated that CACs do form new endocrine cells after β-cell ablation or partial pancreatectomy. We concluded that CACs and their larval predecessors are the same cell type and represent an opportune model to study both β-cell neogenesis and β-cell regeneration. Furthermore, we show that in cftr loss-of-function mutants, there is a deficiency of larval CACs, providing a possible explanation for pancreatic complications associated with cystic fibrosis.

No MeSH data available.