FIGURE

Fig. 3

ID
ZDB-FIG-161018-3
Publication
Lu et al., 2016 - IGFBP1 increases β-cell regeneration by promoting α- to β-cell transdifferentiation
Other Figures
All Figure Page
Back to All Figure Page
Fig. 3

Cellular mechanisms of Igfbp1a′s effect on βcell regeneration

A-C. Tg(ins:GFP);Tg(tp1:H2BmCherry);Tg(ins:FlagNTR) transgenics, with or without Tg(bactin:igfbp1a), were treated with MTZ from 3 to 4 dpf to ablate the β cells and were then allowed to regenerate from 4 to 6 dpf. Representative confocal images at 6 dpf of control (A) and Tg(bactin:igfbp1a) (B) larvae showing a modest number of ins+ tp1+ coexpressing cells, indicated by arrows, after 2 days of regeneration. Scale bars: 15 µm. (C) Quantification of the total number of β cells (green bars) at 6 dpf, and of β cells expressing Tg(tp1:H2BmCherry), that is, of ductal origin (yellow bars). ****< 0.0001, ns= nonsignificant. = 23 larvae in the control group, = 17 larvae in the Tg(bactin:igfbp1a) group.

D-F. Tg(ins:VenuszGeminin) was examined in control and Tg(bactin:igfbp1a) larvae at 6 dpf, after βcell ablation 3-4 dpf by using Tg(ins:FlagNTR). Representative confocal images (D, E) at 6 dpf of control and Tg(bactin:igfbp1a) larvae showing cell cycle activation of β cells in green. Scale bars: 15 µm. (F) Quantification of the total number of β cells with activated cell cycle at 6 dpf, ***< 0.001, = 32 larvae in the control group, = 41 larvae in the Tg(bactin:igfbp1a) group. The number of β cells with activated cell cycle is displayed together with the total number of β cells in experiments with the same setup, ****< 0.0001. = 39 larvae in the control group, = 33 larvae in the Tg(bactin:igfbp1a) group.

G-I. EdU was used as a marker for cell cycle progression. Tg(ins:H2BGFP);Tg(ins:FlagNTR) transgenics, with or without Tg(bactin:igfbp1a), were treated with MTZ from 3 to 4 dpf to ablate their β cells, and subsequently incubated with EdU during regeneration from 4 to 6 dpf. Representative confocal images (G, H) at 6 dpf of control and Tg(bactin:igfbp1a) larvae showing β cells in green and the β cells that had incorporated EdU in yellow (green and red overlap; arrowheads). Scale bars: 20 µm. (I) Quantification of the total number of β cells (green bars) at 6 dpf and of β cells that incorporated EdU (yellow bars) during βcell regeneration from 4 to 6 dpf. **< 0.01, ns = nonsignificant. = 16 in both the control and the Tg(bactin:igfbp1a) group.

J-L. Tg(ins:kaede);Tg(ins:CFPNTR) transgenics, with or without Tg(bactin:igfbp1a), were injected with a control morpholino or a morpholino that knocked down arx, which is necessary for αcell differentiation and thus glucagon expression (J, K). Scale bars: 20 µm. (L) Quantification of the total number of regenerating β cells at 4 dpf (after βcell ablation at 2-3 dpf). ****< 0.0001, *< 0.05, ns=nonsignificant. = 42, 20, 48 and 20, respectively.

Expression Data
Genes:
Antibody:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage: Day 6

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Day 6

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO J.