ZFIN ID: ZDB-FIG-111018-23
Mouillesseaux et al., 2011 - Mutation in utp15 Disrupts Vascular Patterning in a p53-Dependent Manner in Zebrafish Embryos. PLoS One   6(9):e25013 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Prim-15

Fig. 6 Loss of function of utp15 induces p53 activity.

A, Brightfield Phase contrast images of 30 hpf embryos. LA1908 mutant embryos manifest defects in the brain and the vasculature as well as a significantly shorter body when compared to their wild type siblings. Injection of 2 ng p53MO is sufficient to prevent CNS necrosis (A), restore normal CVP morphology (A′), and normalize the gross morphology of the mutant embryos. A′, Magnified image of boxed regions of Tg[kdrl:GFP] embryos in (A). Note normal ISV patterning and presence of EC sprouting within the CVP (white arrows), resulting in a prototypical plexus. B, Chi-squared analysis of WT:Mutant ratios from 3 independently replicated experiments. n = 220 uninjected control embryos, 298 2 ng p53MO-injected embryos, * = p-value<0.001. Data are average +/- SEM. C–E, Analysis of tbx20 expression, a marker of arterial endothelial cells, demonstrated p53MO also restores normal expression levels and patterning (E) which were lost in mutants (D) relative to WT (C) embryos. Embryos are representative of experiments performed in triplicate on a minimum of 5 embryos per condition. F–H, thbs1 is expressed in a p53-dependent manner. Under normal conditions (F), expression is observed in the floor plate, dorsal neural tube, and in the ventral somites/axial vasculature. In mutant embryos, thbs1 expression is substantially upregulated (G). Knockdown of p53 reduced thbs1 expression in a dose-dependent manner. Roughly half (18/41) of the embryos injected with 2 ng p53MO have thbs1 levels as shown in (H), with the remainder (23/41) displaying WT (F) levels. When the dose is increased to 4 ng (I), 82% of embryos (31/38) had WT (F) levels of thbs1 mRNA, while only 18% (7/38) appeared as in (H).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
tbx20 la116Tg standard conditions Prim-5 artery blood vessel endothelial cell ISH
utp15la1908/la1908; la116Tg standard conditions Prim-5 artery blood vessel endothelial cell ISH
utp15la1908/la1908; la116Tg + MO4-tp53 standard conditions Prim-5 artery blood vessel endothelial cell ISH
thbs1b la116Tg standard conditions Prim-5 axial vasculature ISH
Prim-5 floor plate ISH
Prim-5 neural tube dorsal region ISH
Prim-5 somite ventral region ISH
utp15la1908/la1908; la116Tg standard conditions Prim-5 axial vasculature ISH
Prim-5 floor plate ISH
Prim-5 neural tube dorsal region ISH
Prim-5 somite ventral region ISH
utp15la1908/la1908; la116Tg + MO4-tp53 standard conditions Prim-5 axial vasculature ISH
Prim-5 floor plate ISH
Prim-5 neural tube dorsal region ISH
Prim-5 somite ventral region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
utp15la1908/la1908; la116Tg standard conditions Prim-5 negative regulation of angiogenesis increased occurrence, abnormal
Prim-15 caudal vein plexus complexity, abnormal
Prim-15 caudal vein plexus decreased thickness, abnormal
Prim-15 caudal vein plexus endothelial tip cell decreased amount, abnormal
Prim-15 central nervous system necrotic, abnormal
Prim-15 intersegmental vessel decreased amount, abnormal
Prim-15 intersegmental vessel hypoplastic, abnormal
Prim-15 sprouting angiogenesis delayed, abnormal
Prim-15 whole organism anterior-posterior axis decreased length, abnormal
utp15la1908/la1908; la116Tg + MO4-tp53 standard conditions Prim-5 regulation of angiogenesis process quality, normal
Prim-15 caudal vein plexus morphology, normal
Prim-15 central nervous system structure, normal
Prim-15 intersegmental vessel morphology, normal
Prim-15 whole organism morphology, normal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One