ZFIN Image: Andersson et al., 2012, Fig. 4

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Figure Caption

Fig. 4

NECA Enhances Proliferation of β Cells, and Not Cell Proliferation in General

(A–C) Proliferation of α cells was assessed by EdU incorporation in Tg(gcg:GFP)-expressing α cells. Tg(gcg:GFP);Tg(ins:flag-NTR) larvae were treated with MTZ from 3-4 dpf for ablation of the β cells and subsequently treated with DMSO/NECA and EdU during β cell regeneration from 4-6 dpf. (A) Confocal image of a DMSO-treated larva displaying one α cell that had incorporated EdU (arrow). (B) Confocal image of a NECA-treated larva that does not show any EdU incorporation in α cells, but substantial EdU incorporation in β cells (positive for Tg(ins:flag-NTR) expression). (C) Quantification of EdU incorporation in α cells (green bars) and β cells (red bars) in the presence of DMSO or NECA. Note that the number of α cells that incorporated EdU during treatment with NECA did not significantly increase, although incorporation of EdU in Tg(ins:flag-NTR)-expressing β cells increased markedly in the same larvae. n = 11 larvae for each group.

(D–F) Proliferation of δ cells was assessed by EdU incorporation in Tg(sst:RFP)-expressing δ cells. Tg(sst:RFP);Tg(ins:flag-NTR) larvae were treated with MTZ from 3-4 dpf for ablation of the β cells and subsequently treated with DMSO/NECA and EdU during β cell regeneration from 4-6 dpf. (D) Confocal image of a DMSO-treated larva displaying one δ cell that had incorporated EdU (arrow). (E) Confocal image of a NECA-treated larva displaying one δ cell that had incorporated EdU, and substantial EdU incorporation in β cells (positive for Tg(ins:flag-NTR) expression). (F) Quantification of EdU incorporation in δ cells (red bars) and β cells (green bars) in the presence of DMSO or NECA. n = 10 larvae for each group.

(G) Quantification of proliferation in the gut and liver in Tg(ins:CFP-NTR) larvae following β cell ablation from 3-4 dpf and subsequent treatment with DMSO/NECA from 4-6 dpf. The percentage of cells that incorporated EdU between 4-6 dpf during β cell regeneration did not change with NECA treatment when compared to DMSO-treated controls. n = 10 larvae for each group.

(H) The mitosis marker Phospho-Histone H3 (P-H3) colabels a β cell that expresses both insulin and Tg(neurod:GFP) (arrow) following β cell ablation from 3-4 dpf and subsequent treatment with NECA from 4-6 dpf. Note that the β cell that is positive for P-H3 is located in the periphery of the islet.

(I) Quantification of β cell proliferation, as marked by colocalization of insulin and P-H3, following β cell ablation from 3-4 dpf and subsequent treatment with DMSO/NECA from 4-6 dpf. n = 42 larvae for DMSO treated; n = 48 larvae for NECA treated. Error bars represent SEM. See also Figures S4 and S5.

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Acknowledgments

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Reprinted from Cell Metabolism, 15(6), Andersson, O., Adams, B.A., Yoo, D., Ellis, G.C., Gut, P., Anderson, R.M., German, M.S., and Stainier, D.Y., Adenosine Signaling Promotes Regeneration of Pancreatic beta Cells In Vivo, 885-894, Copyright (2012) with permission from Elsevier. Full text @ Cell Metab.