El Ouaamari et al., 2016 - SerpinB1 Promotes Pancreatic β Cell Proliferation. Cell Metabolism   23(1):194-205 Full text @ Cell Metab.

Fig. 3

Overexpression of Serpinb1 in Zebrafish Enhances β Cell Regeneration and Proliferation

(A) Schematic of experimental plan.

(B–E) Representative images at 6 dpf of Tg(ins:kaede);Tg(ins:CFP-NTR) transgenic larvae that had not been injected (control), or were injected at the 1-cell stage with transposase mRNA + bactin:H2A-mCherry, bactin:serpina7, or bactin:serpinb1; were subjected to β cell ablation by metronidazole during 3–4 dpf; and were subsequently allowed to regenerate for 2 days.

(F) Quantification of β cell regeneration at 6 dpf in control (n = 87), bactin:H2A-mCherry-overexpressing (n = 61), bactin:serpina7-overexpressing (n = 46), and bactin:serpinb1-overexpressing (n = 36) Tg(ins:kaede);Tg(ins:CFP-NTR) larvae.

(G–J) Control (n = 27) and bactin:serpinb1-overexpressing (n = 18) Tg(ins:H2B-GFP);Tg(ins:Flag-NTR) transgenics were treated with metronidazole from 3 to 4 dpf to ablate the β cells and subsequently incubated with EdU during regeneration from 4 to 6 dpf. Representative confocal images at 6 dpf of control (G) and bactin:serpinb1-overexpressing (H) larvae showing β cells in green and the β cells that had incorporated EdU in yellow (green and red overlap; arrowheads). (I) Quantification of the total number of β cells at 6 dpf. (J) Quantification of β cells that incorporated EdU during β cell regeneration from 4 to 6 dpf.

(K–N) To determine whether Serpinb1 affects β cell proliferation during regular development, we treated control (n = 25) and bactin:serpinb1-overexpressing Tg(ins:H2B-GFP) (n = 21) transgenic larvae with EdU from 4 to 6 dpf. Representative confocal images at 6 dpf of control (K) and bactin:serpinb1-overexpressing (L) larvae showing β cells in green and the β cells that had incorporated EdU in yellow (green and red overlap; arrowhead). (M) Quantification of the total number of β cells at 6 dpf. (N) Quantification of β cells that incorporated EdU from 4 to 6 dpf. Data shown are the mean ± SEM; ****p < 0.0001, **p < 0.01, *p < 0.05. Arrows indicate proliferating cells.

EXPRESSION / LABELING:
Gene:
Fish:
Condition:
Anatomical Term:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Day 4
Acknowledgments:
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Reprinted from Cell Metabolism, 23(1), El Ouaamari, A., Dirice, E., Gedeon, N., Hu, J., Zhou, J.Y., Shirakawa, J., Hou, L., Goodman, J., Karampelias, C., Qiang, G., Boucher, J., Martinez, R., Gritsenko, M.A., De Jesus, D.F., Kahraman, S., Bhatt, S., Smith, R.D., Beer, H.D., Jungtrakoon, P., Gong, Y., Goldfine, A.B., Liew, C.W., Doria, A., Andersson, O., Qian, W.J., Remold-O'Donnell, E., Kulkarni, R.N., SerpinB1 Promotes Pancreatic β Cell Proliferation, 194-205, Copyright (2016) with permission from Elsevier. Full text @ Cell Metab.