ZFIN ID: ZDB-FIG-161018-4
Lu et al., 2016 - IGFBP1 increases β-cell regeneration by promoting α- to β-cell transdifferentiation. The EMBO journal   35(18):2026-44 Full text @ EMBO J.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Conditions:
Anatomical Terms:
Stage: Day 6
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Day 6

Fig. 4

Igfbp1a increases βcell regeneration by potentiating α to βcell transdifferentiation

A-E. Control and Tg(bactin:igfbp1a)overexpressing larvae were treated with MTZ from 3 to 4 dpf to ablate β cells and analyzed at 6 dpf, after 2 days of regeneration. Representative confocal images (A, B) of control and Tg(bactin:igfbp1a)overexpressing Tg(ins:dsRed);Tg(gcg:GFP);Tg(ins:FlagNTR) larvae. A bihormonal glucagon and insulinexpressing cell (gcg+;ins+) is indicated by an arrow. Representative confocal images (C, D) of control and Tg(bactin:igfbp1a)overexpressing Tg(gcg:GFP);Tg(ins:FlagNTR) larvae stained for Pdx1. Pdx1 and glucagonexpressing cells (pdx1+;gcg+) are indicated by arrows. Scale bars: 20 µm. (E) Quantification of gcg+;ins+ and pdx1+;gcg+ cells in the control and Tg(bactin:igfbp1a) group; ***< 0.001, **= 0.0012; = 18 and 18 in the control groups, = 15 and 10 in the Tg(bactin:igfbp1a) groups, for the gcg+;ins+ and pdx1+;gcg+ quantification, respectively.

F-I. Lineagetracing evidence supports α to βcell transdifferentiation. (F) Schema for temporal and spatial lineage tracing. loxPmediated excision of the STOP cassette permits heatinducible expression of the stable fusionprotein H2BGFP. Control and bactin:igfbp1aoverexpressing Tg(hsp:loxPmCherrySTOPloxPH2BGFP);Tg(gcg:CRE);Tg(ins:FlagNTR) larvae were first heatshocked at 3 dpf to label the glucagonexpressing cells with the stable fluorescent protein H2BGFP, thereafter treated with MTZ from 3 to 4 dpf to ablate the β cells, and analyzed at 6 dpf after 2 days of regeneration. Note that the temporal control of the lineage tracing is mediated by a heatshock. (G) Quantification of insulinexpressing cells that originate from α cells. Control larvae, n = 13; bactin:igfbp1a larvae, = 16. **= 0.0051. Representative confocal images (H, I) at 6 dpf (after 2 days of regeneration) of control and bactin:igfbp1aoverexpressing larvae stained for insulin, glucagon, topro, and H2BGFP. ins+;H2BGFP+ cells are indicated by the arrowheads, and H′ and I′ show staining against only insulin and H2BGFP. Moreover, most ins+;H2BGFP+ cells no longer express glucagon. Scale bars: 15 µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed ia1Tg; m1018Tg; s950Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 regenerating tissue pancreatic B cell IFL
ki103Tg ; m1018Tg ; ia1Tg ; s950Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 regenerating tissue pancreatic B cell IFL
GFP ia1Tg ; s950Tg ; ki103Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 glucagon secreting cell IHC
Day 6 regenerating tissue pancreatic B cell IHC
ia1Tg; m1018Tg; s950Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 glucagon secreting cell IFL
Day 6 regenerating tissue pancreatic B cell IFL
ki103Tg ; m1018Tg ; ia1Tg ; s950Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 glucagon secreting cell IFL
Day 6 regenerating tissue pancreatic B cell IFL
s923Tg; s950Tg; s962Tg heat shock, chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 glucagon secreting cell IHC
Day 6 regenerating tissue pancreatic B cell IHC
s950Tg ; ia1Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 glucagon secreting cell IHC
Day 6 regenerating tissue pancreatic B cell IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab4-ins IHC s923Tg; s950Tg; s962Tg heat shock, chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 regenerating tissue pancreatic B cell
Phenotype Details
Fish Conditions Stage Phenotype
ki103Tg ; m1018Tg ; ia1Tg ; s950Tg chemical ablation: pancreatic B cell, chemical treatment by environment: metronidazole Day 6 pancreatic B cell regeneration increased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal The EMBO journal for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ EMBO J.