ZFIN ID: ZDB-FIG-180827-49
Zhao et al., 2018 - Deficiency in class III PI3-kinase confers postnatal lethality with IBD-like features in zebrafish. Nature communications   9:2639 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Day 6 to Days 7-13
PHENOTYPE:
Fish:
Observed In:
Stage: Days 7-13

Fig. 3

Pro-inflammatory responses in the intestine of pik3c3 mutants. a Heatmap of inflammation-related genes shows the gradual stimulation of inflammatory response from 6 to 8 dpf in the mutants. b qRT-PCR analysis of the indicated inflammation-related genes. cxcl8a is the zebrafish homolog of IL8. Experiments were performed and data analyzed as described in Fig. 2b. c mpx:GFP is a transgenic zebrafish line with neutrophils labeled by GFP. In 8 dpf mutants, neutrophils mainly accumulate in the digestive tract. Scale bar, 500 µm. d High magnification confocal imaging of (c). Scale bar, 50 µm. e Cross sections of intestinal bulb in D8 embryos. Neutrophil infiltration of intestinal epithelium is clearly seen in the mutants. Scale bar, 50 µm. f mpeg1:GFP-labeled macrophage lineage cells appear normal in the mutants. Scale bar, 500 µm. g qRT-PCR analysis of the expression of IBD candidate genes in dissected intestine of pik3c3 mutants at 8 dpf. Inflammatory response genes (il1b, card9, mmp9) and igfbp1a (one of the homologs of human IGFBP1, a gene related to the prognosis of CD) are significantly up-regulated in the mutants. Bacteria sensing and autophagy-related genes (nod2, atg16l1, and irgm) and tnfa are not stimulated. Barrier function-related genes hnf4a and ttc7a are severely down-regulated in the mutants while the hp gene, which functions to open up tight junctions, is up-regulated in the mutants. actb1 is used as internal control and data represent mean ± SD from three biological repeats. p-values are determined by unpaired two-tailed Student’s t-test (NS, non-significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
atg16l1 TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
card9 TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 whole organism RTPCR
cxcl8a TU standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 Not Detected whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 whole organism RTPCR
EGFP rj30Tg standard conditions Days 7-13 intestine neutrophil IFL
rj30Tg ; pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine neutrophil IFL
rj30Tg ; pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine neutrophil IFL
hnf4a TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
hp TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
igfbp1a TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
igfbp1b TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
il1b TU standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 Not Detected whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
lect2.1 TU standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 Not Detected whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 whole organism RTPCR
mmp9 TU standard conditions Day 6 whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
mpx TU standard conditions Day 6 whole organism RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 whole organism RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 whole organism RTPCR
Days 7-13 whole organism RTPCR
nod2 TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
tnfa TU standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 Not Detected whole organism RTPCR
pik3c3zf2160/zf2160 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
pik3c3zf2161/zf2161 standard conditions Day 6 Not Detected whole organism RTPCR
Days 7-13 intestine RTPCR
Days 7-13 whole organism RTPCR
ttc7a TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
zmp:0000000951 TU standard conditions Days 7-13 intestine RTPCR
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine RTPCR
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestine hnf4a expression decreased amount, abnormal
Days 7-13 intestine hp expression increased amount, abnormal
Days 7-13 intestine igfbp1a expression increased amount, abnormal
Days 7-13 intestine il1b expression increased amount, abnormal
Days 7-13 intestine mmp9 expression increased amount, abnormal
Days 7-13 intestine ttc7a expression decreased amount, abnormal
Days 7-13 whole organism card9 expression increased amount, abnormal
Days 7-13 whole organism cxcl8a expression increased amount, abnormal
Days 7-13 whole organism il1b expression increased amount, abnormal
Days 7-13 whole organism lect2.1 expression increased amount, abnormal
Days 7-13 whole organism mmp9 expression increased amount, abnormal
Days 7-13 whole organism mpx expression increased amount, abnormal
Days 7-13 whole organism tnfa expression increased amount, abnormal
pik3c3zf2160/zf2160 ; ihb20Tg standard conditions Days 7-13 macrophage normal amount, normal
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestine hnf4a expression decreased amount, abnormal
Days 7-13 intestine hp expression increased amount, abnormal
Days 7-13 intestine igfbp1a expression increased amount, abnormal
Days 7-13 intestine il1b expression increased amount, abnormal
Days 7-13 intestine mmp9 expression increased amount, abnormal
Days 7-13 intestine ttc7a expression decreased amount, abnormal
Days 7-13 whole organism card9 expression increased amount, abnormal
Days 7-13 whole organism cxcl8a expression increased amount, abnormal
Days 7-13 whole organism il1b expression increased amount, abnormal
Days 7-13 whole organism lect2.1 expression increased amount, abnormal
Days 7-13 whole organism mmp9 expression increased amount, abnormal
Days 7-13 whole organism mpx expression increased amount, abnormal
Days 7-13 whole organism tnfa expression increased amount, abnormal
pik3c3zf2161/zf2161 ; ihb20Tg standard conditions Days 7-13 macrophage normal amount, normal
rj30Tg ; pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelium has extra parts of type neutrophil, abnormal
Days 7-13 intestine neutrophil EGFP expression increased amount, abnormal
rj30Tg ; pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelium has extra parts of type neutrophil, abnormal
Days 7-13 intestine neutrophil EGFP expression increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Nature communications for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Nat. Commun.