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ZFIN ID: ZDB-FIG-160823-1
Kuusela et al., 2016 - Tankyrase inhibition aggravates kidney injury in the absence of CD2AP. Cell Death & Disease   7:e2302 Full text @ Cell Death Dis.
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Condition:
Knockdown Reagent:
Anatomical Term:
Stage: Day 5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Day 5

Fig. 7

Simultaneous disruption of cd2ap expression and tankyrase activity aggravates kidney injury in zebrafish larvae. (a and b) Knockdown of cd2ap with morpholino antisense oligonucleotides (cd2ap-MO) in zebrafish causes pericardial edema (arrowhead) at 5 days post fertilization (dpf) indicative of kidney dysfunction (b). Control-MO (C-MO)-injected larvae appear developmentally normal (a). (c and d) Inhibition of tankyrase activity with XAV939 in the cd2ap morphant between 3 and 5 dpf induces yolk sac edema and more severe pericardial edema (d, arrowhead) than knockdown of cd2ap alone (b), whereas XAV939 treatment has no effect on the control morphant (c). (e and g) Histological sections of 5 dpf control morphant (e) and control morphant treated with XAV939 (g) show normal glomeruli (arrowheads) and proximal convoluted tubules (arrows). (f) cd2ap morphant shows a disturbed glomerular structure (arrowhead) with cysts (asterisk). (h) Inhibition of tankyrase activity with XAV939 in cd2ap morphants leads to even more destructed glomerular structure with a dilated (asterisk) and stretched septal glomerulus in the midline (arrowhead) and dilated proximal convoluted tubule (arrow). (i) Immunoblotting for podocin and active β-catenin (ABC) in control morphants (C-MO) and cd2ap morphants (cd2ap-MO) with and without XAV939 treatment. α-Tubulin is included as a control. (j) Quantification of three replicate blots as in (i) shows significant downregulation of podocin and upregulation of ABC in cd2ap morphants compared with control morphants. XAV939 treatment further downregulates podocin in cd2ap morphants. (k) Knockdown of cd2ap with cd2ap-MO in zebrafish leads to an increase in total PARylation at 5 dpf compared with control-MO treated zebrafish. P-values were calculated with one-way ANOVA (**P<0.01; ***P<0.001). Scale bar: (e-h) 25 ยตm

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ctnnb1 Ab3-ctnnb1 WT control Day 5 whole organism WB
Ab3-ctnnb1 WT + MO1-cd2ap standard conditions Day 5 whole organism WB
Ab3-ctnnb1 WT + MO1-cd2ap chemical treatment: XAV939 Day 5 whole organism WB
nphs2 Ab2-nphs2 WT control Day 5 whole organism WB
Ab2-nphs2 WT + MO1-cd2ap standard conditions Day 5 whole organism WB
Ab2-nphs2 WT + MO1-cd2ap chemical treatment: XAV939 Day 5 whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-cd2ap standard conditions Day 5 pericardium edematous, abnormal
Day 5 pronephric glomerular capillary distended, abnormal
Day 5 pronephric glomerulus morphology, abnormal
Day 5 protein poly-ADP-ribosylation increased occurrence, abnormal
Day 5 whole organism nphs2 expression decreased amount, abnormal
Day 5 whole organism ctnnb1 expression increased amount, abnormal
WT + MO1-cd2ap chemical treatment: XAV939 Day 5 pericardium edematous, abnormal
Day 5 pronephric capsular space dilated, abnormal
Day 5 pronephric glomerulus cystic, abnormal
Day 5 pronephric glomerulus dilated, abnormal
Day 5 proximal convoluted tubule dilated, abnormal
Day 5 whole organism nphs2 expression decreased amount, abnormal
Day 5 whole organism ctnnb1 expression increased amount, abnormal
Day 5 yolk syncytial layer edematous, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Death & Disease for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Death Dis.