ZFIN ID: ZDB-FIG-170807-7
Wu et al., 2017 - tomm22 Knockdown-Mediated Hepatocyte Damages Elicit Both the Formation of Hybrid Hepatocytes and Biliary Conversion to Hepatocytes in Zebrafish Larvae.. Gene Expression   17(3):237-249 Full text @ Gene Expr.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Day 5 to Days 7-13
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagent:
Observed In:
Stage Range: Day 5 to Days 7-13

Fig. 6

Macrophage ablation impairs the formation of hybrid hepatocytes in tomm22 MO-injected larvae. (A) Confocal single-optical section images showing the expression of mpeg1:Dendra2 (green, macrophages) and fabp10a:CFP-NTR (blue, hepatocytes) in the liver at 5 dpf. Arrows point to macrophages. (B) Confocal single-optical section images showing the expression of mpeg1:Dendra2 (green), Tp1:H2B-mCherry (red), and fabp10a:CFP-NTR (blue) in the liver at 7 dpf. Arrows point to macrophages in the control liver; arrowheads point to a macrophage engulfing a hepatocyte in tomm22 MO-injected larvae. (C) Fluorescence images showing macrophage NTR-mCherry expression before and after metronidazole (Mtz) treatment. Mtz (10 mM) was treated from 4 to 7 dpf. Note the great reduction in the number of NTR-mCherry+ macrophages in the Mtz-treated larvae at 7 dpf compared with the DMSO-treated controls. Lateral views, dorsal up, anterior to the left. Scale bar: 250 μm. (D) Scheme illustrating the period of Mtz treatment and harvest stage for macrophage ablation for (E–I). (E) Confocal single-optical section images showing the expression of Tp1:H2B-mCherry (red) and Hnf4a (gray) in the liver at 7 dpf. Arrows point to H2B-mCherry/Hnf4a double-positive cells; arrowheads point to Hnf4a single-positive cells. (F) Graph showing the percentage of H2B-mCherry cells among Hnf4a+ cells. Red marks indicate the larvae shown in (E). (G) Confocal single-optical section images showing the expression of fabp10a:mAGFP-gmnn (green), Tp1:H2B-mCherry (red), and Hnf4a (gray) in the liver at 7 dpf. Arrows point to mAGFP-gmnn/H2B-mCherry/Hnf4a triple-positive cells; arrowheads point to mAGFP-gmnn+/H2B-mCherry/Hnf4a+ cells. (H) Graph showing the percentage of fabp10a:mAGFP-gmnn+ cells among Hnf4a+ cells. Red marks indicate the larvae shown in (G). (I) Graph showing the percentage of H2B-mCherry+ or H2B-mCherry proliferating Hnf4a+ cells. Red marks indicate the larvae shown in (G). n indicates the number of larvae examined. (J) Confocal single-optical section images showing the expression of WRE:d2GFP (green, Wnt activity) and Tp1:H2B-mCherry (red) in the liver at 6 dpf. Anti-GFP antibody was used to reveal WRE:d2GFP expression. Note the complete absence of hepatic d2GFP expression in the XAV939-treated, tomm22 MO-injected larvae, whereas its faint expression in macrophage-ablated, tomm22 MO-injected larvae. Scale bars: 20 μm, except (C); error bars: ±SEM.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
CFP s931Tg; s939Tg; uwm12Tg control Days 7-13 hepatocyte IHC
s931Tg; s939Tg; uwm12Tg + MO1-tomm22 standard conditions Days 7-13 hepatocyte IHC
s931Tg; uwm12Tg control Day 5 hepatocyte IHC
s931Tg; uwm12Tg + MO1-tomm22 standard conditions Day 5 hepatocyte IHC
d2EGFP c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 control Day 6 liver IHC
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Day 6 liver IHC
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical treatment by environment: XAV939 Day 6 liver IHC
Dendra2 s931Tg; s939Tg; uwm12Tg control Days 7-13 liver macrophage IHC
s931Tg; s939Tg; uwm12Tg + MO1-tomm22 standard conditions Days 7-13 liver macrophage IHC
s931Tg; uwm12Tg control Day 5 liver macrophage IHC
s931Tg; uwm12Tg + MO1-tomm22 standard conditions Day 5 liver macrophage IHC
mAGFP c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 control Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
mCherry c264Tg; gl24Tg control Days 7-13 macrophage IHC
c264Tg; gl24Tg chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 Not Detected macrophage IHC
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 control Day 6 liver IHC
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Day 6 liver IHC
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical treatment by environment: XAV939 Day 6 liver IHC
c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 control Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
c264Tg; gl24Tg; s939Tg + MO1-tomm22 control Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
c264Tg; gl24Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 cholangiocyte IHC
Days 7-13 hepatocyte IHC
s931Tg; s939Tg; uwm12Tg control Days 7-13 cholangiocyte IHC
s931Tg; s939Tg; uwm12Tg + MO1-tomm22 standard conditions Days 7-13 cholangiocyte IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
c264Tg; gl24Tg chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 macrophage decreased amount, abnormal
Days 7-13 macrophage mCherry expression absent, abnormal
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 control Day 6 liver d2EGFP expression increased amount, abnormal
Day 6 liver Wnt signaling pathway increased occurrence, abnormal
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Day 6 liver d2EGFP expression amount, ameliorated
Day 6 liver Wnt signaling pathway occurrence, ameliorated
c264Tg; gl24Tg; kyu1Tg; s939Tg + MO1-tomm22 chemical treatment by environment: XAV939 Day 6 liver d2EGFP expression amount, ameliorated
Day 6 liver Wnt signaling pathway occurrence, ameliorated
c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 control Days 7-13 cholangiocyte mAGFP expression increased amount, abnormal
Days 7-13 hepatocyte mCherry expression increased amount, abnormal
c264Tg; gl24Tg; pt608Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 cholangiocyte mAGFP expression increased amount, abnormal
Days 7-13 hepatocyte mCherry expression increased amount, abnormal
c264Tg; gl24Tg; s939Tg + MO1-tomm22 control Days 7-13 hepatocyte mCherry expression increased amount, abnormal
c264Tg; gl24Tg; s939Tg + MO1-tomm22 chemical ablation: macrophage, chemical treatment by environment: metronidazole Days 7-13 hepatocyte mCherry expression increased amount, abnormal
s931Tg; s939Tg; uwm12Tg + MO1-tomm22 standard conditions Days 7-13 liver macrophage increased amount, abnormal
Days 7-13 liver macrophage shape, abnormal
s931Tg; uwm12Tg + MO1-tomm22 standard conditions Day 5 liver macrophage increased amount, abnormal
Acknowledgments:
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