ZFIN ID: ZDB-FIG-160316-23
Xu et al., 2016 - Four and a Half LIM Domains 1b (Fhl1b) Is Essential for Regulating the Liver versus Pancreas Fate Decision and for β-Cell Regeneration. PLoS Genetics   12:e1005831 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-15 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Long-pec

Fig. 3

Decreased Fhl1b activity augments induction of pancreatic endocrine cells.

(A-H) Confocal images showing Tg(P0-pax6b:GFP)ulg515 (A and B, green), Insulin (C and D, red), Somatostatin (E and F, grey), and Glucagon (G and H, blue) expression at 30 hpf, comparing control embryos (A, C, E, and G) and fhl1b morphants (B, D, F, and H). The number of Tg(P0-pax6b:GFP)ulg515 (B)- and Insulin (D)-expressing cells was significantly increased in fhl1b morphants compared to that of control embryos (A and C, respectively). The number of Somatostatin-expressing cells was also increased (F), while that of Glucagon-expressing cells appeared unaffected (H) in fhl1b morphants compared to that of control embryos (E and G, respectively). (I) Quantification of the number (mean±SD) of total and individual pancreatic endocrine hormone-expressing cells, comparing that of control embryos and fhl1b morphants at 30 hpf. The absolute number of Tg(P0-pax6b:GFP)ulg515-, Insulin-, and Somatostatin-expressing cells was increased from 82.6±4.5, 30.6±1.5, and 20.7±0.8, respectively, in control embryos, to 103±2.0, 44.6±2.0, and 26.7±2.0, respectively, in fhl1b morphants, while that of Glucagon-expressing cells appeared unaffected (26.5±0.7 cells in controls vs. 24.6±1.5 cells in fhl1b morphants). Cells in 20 planes of confocal images from 5 individual embryos were counted. Asterisks indicate statistical significance: **, P < 0.01; ***, P < 0.001. (J-K′) Confocal images of Tg(ins:GFP)zf5;Tg(ins:dsRed)m1018 control embryos (J and J′) and fhl1b morphants (K and K′) at 48 hpf. Compared with the control embryos (J), fhl1b morphants showed an increased number of GFP-only-positive β-cells (K, white arrows). (L) Quantification of the number (mean±SD) of GFP- and dsRed-double positive (yellow) and GFP-only-positive (green) β-cells, comparing that of control embryos and fhl1b morphants at 48 hpf. In control embryos, 5.0±0.7 β-cells were GFP-only-positive, while 12.2±2.3 β-cells were GFP-only-positive in fhl1b morphants. Cells in 20 planes of confocal images from 5 individual embryos were counted. Asterisks indicate statistical significance: ***, P < 0.001. A-H and J-K′, confocal projection images, ventral views, anterior to the top. Scale bars, 20 µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed zf5Tg ; m1018Tg control Long-pec insulin secreting cell IHC
zf5Tg ; m1018Tg + MO1-fhl1b + MO2-fhl1b standard conditions Long-pec insulin secreting cell IHC
GFP ulg515Tg control Prim-15 pancreatic bud endocrine cell IHC
ulg515Tg + MO1-fhl1b + MO2-fhl1b standard conditions Prim-15 pancreatic bud endocrine cell IHC
zf5Tg ; m1018Tg control Long-pec insulin secreting cell IHC
zf5Tg ; m1018Tg + MO1-fhl1b + MO2-fhl1b standard conditions Long-pec insulin secreting cell IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-isl IHC WT control Prim-15 insulin secreting cell
IHC WT + MO1-fhl1b + MO2-fhl1b standard conditions Prim-15 insulin secreting cell
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-fhl1b + MO2-fhl1b standard conditions Prim-15 endocrine pancreas pancreas induction increased occurrence, abnormal
Prim-15 insulin secreting cell ab2-isl labeling increased distribution, abnormal
Prim-15 insulin secreting cell increased amount, abnormal
Prim-15 somatostatin secreting cell increased amount, abnormal
ulg515Tg + MO1-fhl1b + MO2-fhl1b standard conditions Prim-15 endocrine pancreas pancreas induction increased occurrence, abnormal
Prim-15 pancreas endocrine cell increased amount, abnormal
Prim-15 pancreatic bud endocrine cell GFP expression increased distribution, abnormal
zf5Tg ; m1018Tg + MO1-fhl1b + MO2-fhl1b standard conditions Long-pec endocrine pancreas pancreas induction increased occurrence, abnormal
Long-pec insulin secreting cell DsRed expression increased distribution, abnormal
Long-pec insulin secreting cell GFP expression increased distribution, abnormal
Long-pec insulin secreting cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.