ZFIN ID: ZDB-FIG-160205-113
Khaliq et al., 2015 - Id2a is required for hepatic outgrowth during liver development in zebrafish. Mechanisms of Development   138 Pt 3:399-414 Full text @ Mech. Dev.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-25 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-25 to Protruding-mouth

Fig. 3

id2a knockdown reduces liver size but does not block hepatoblast specification or hepatocyte differentiation. (A) Epifluorescence images revealing a decreased number of Tp1:GFP+ BECs in the liver of id2a MO-injected embryos at 72 hpf compared with controls (squares). Higher magnification images of the square regions are shown in insets. Lateral view, anterior to the left. (B) Confocal projection images revealing fabp10a:dsRed (hepatocytes; red), Tp1:GFP (BECs; green) and Anxa4 (the hepatopancreatic ductal system; blue) expression (Zhang et al., 2014). In id2a MO-injected embryos, liver size was greatly reduced and intrahepatic BECs appeared aggregated, displaying a branching defect. (C–H) id2a MO-injected and uninjected control embryos were processed for WISH with hhex (C), prox1a (D), fabp10a (E), cp (F), sepp1b (G), and cdx1b (H) probes. Overall liver size was greatly reduced in id2a MO-injected embryos as revealed by the hepatoblast markers (hhex and prox1a) and the hepatocyte markers (fabp10a, cp, and sepp1b). However, the expression of these genes was clearly detected in the MO-injected embryos (C–G, arrows), indicating normal hepatoblast specification and hepatocyte differentiation upon id2a knockdown. The induction of the intestinal bulb as assessed by cdx1b expression appeared normal in id2a MO-injected embryos at 36 hpf; however, the intestinal bulb failed to grow at 48 hpf (H, brackets). The percentage of id2a MO-injected embryos exhibiting the representative phenotype shown is indicated in the upper left corner (n = 10–20). The remaining percentage of embryos exhibited an intermediate liver/intestinal bulb phenotype: their liver/intestinal bulb size was still smaller than that of the control embryos. Arrows point to the liver. Scale bar: 250 (A), 20 (B), and 100 (C–H) µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
cdx1b WT control Prim-25 intestinal bulb primordium ISH
Long-pec intestinal bulb ISH
WT + MO1-id2a standard conditions Prim-25 intestinal bulb primordium ISH
Long-pec intestinal bulb ISH
cp WT control Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
WT + MO1-id2a standard conditions Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
DsRed gz15Tg; um14Tg control Protruding-mouth hepatocyte IHC
gz15Tg; um14Tg + MO1-id2a standard conditions Protruding-mouth hepatocyte IHC
EGFP um14Tg ; jh1Tg standard conditions Protruding-mouth liver IFL
um14Tg ; jh1Tg + MO1-id2a standard conditions Protruding-mouth Not Detected liver IFL
fabp10a WT control Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
WT + MO1-id2a standard conditions Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
GFP gz15Tg; um14Tg control Protruding-mouth intrahepatic bile duct epithelial cell IHC
gz15Tg; um14Tg + MO1-id2a standard conditions Protruding-mouth intrahepatic bile duct epithelial cell IHC
hhex WT control Prim-25 hepatoblast ISH
Long-pec hepatoblast ISH
WT + MO1-id2a standard conditions Prim-25 hepatoblast ISH
Long-pec hepatoblast ISH
prox1a WT control Prim-25 hepatoblast ISH
Long-pec hepatoblast ISH
WT + MO1-id2a standard conditions Prim-25 hepatoblast ISH
Long-pec hepatoblast ISH
selenop2 WT control Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
WT + MO1-id2a standard conditions Long-pec hepatocyte ISH
Protruding-mouth hepatocyte ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab-2F11 IHC gz15Tg; um14Tg control Protruding-mouth intrahepatic bile duct
IHC gz15Tg; um14Tg + MO1-id2a standard conditions Protruding-mouth intrahepatic bile duct
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-id2a standard conditions Prim-25 liver decreased size, abnormal
Long-pec intestinal bulb decreased size, abnormal
Long-pec liver decreased size, abnormal
Protruding-mouth liver decreased size, abnormal
gz15Tg; um14Tg + MO1-id2a standard conditions Protruding-mouth intrahepatic bile duct aggregated, abnormal
Protruding-mouth intrahepatic bile duct decreased branchiness, abnormal
Protruding-mouth intrahepatic bile duct epithelial cell GFP expression decreased amount, abnormal
Protruding-mouth intrahepatic bile duct epithelial cell GFP expression spatial pattern, abnormal
Protruding-mouth liver decreased size, abnormal
um14Tg ; jh1Tg + MO1-id2a standard conditions Protruding-mouth liver EGFP expression absent, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Mechanisms of Development for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Mechanisms of Development, 138 Pt 3, Khaliq, M., Choi, T.Y., So, J., Shin, D., Id2a is required for hepatic outgrowth during liver development in zebrafish, 399-414, Copyright (2015) with permission from Elsevier. Full text @ Mech. Dev.