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ZFIN ID: ZDB-FIG-150224-15
Kjaer-Sorensen et al., 2014 - Papp-a2 modulates development of cranial cartilage and angiogenesis in zebrafish embryos. Journal of Cell Science   127(23):5027-37 Full text @ J. Cell Sci.
ADDITIONAL FIGURES
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 3

papp-a2 knockdown causes ventral curvature and notochord defects. (A,B) RT-PCR analysis of the effect of papp-a2-targeted morpholino ei1i on papp-a2 pre-mRNA splicing showing a fast-migrating band (LMW) (A) resulting from the use of a cryptic splice donor site in exon 1, and a slow-migrating band (HMW) (B) resulting from the retention of intron 1 as determined by sequencing. These mis-spliced transcripts encode amino acid residue 1–242 or 1–275 of the prepro-protein plus eight or 30 residues, respectively, encoded by intron 1 followed by a stop codon. Values for e1i1 are the amount of morpholino in ng per embryo. WT, wild type. (C) RT-PCR analysis of the effect of papp-a2-targeted morpholino i3e4 on papp-a2 pre-mRNA splicing. Values for i3e4 are the amount of morpholino in ng per embryo. (D) 24 hpf embryos injected with 5.0ng control morpholino (cMO), 5.0ng papp-a2-targeted splice inhibiting morpholino (e1i1), 2.5ng translation inhibiting morpholino (ATG), or 2.0ng splice-inhibiting morpholino (i3e4) with or without 5ng p53-targeted morpholino (p53). Necrosis of the central nervous system was observed from both e1i1 and ATG, indicating non-specific p53-activating activity (Ekker and Larson, 2001; Robu et al., 2007). Co-injection with p53-targeting morpholino (Robu et al., 2007) specifically diminished the observed necrosis, whereas the other phenotypes remained unaffected. p53-targeting morpholino was included in all subsequent knockdown experiments. Insets show a magnified in view from the area indicated by the arrow. Notochord outlines are indicated with dashed lines. (E) noto whole mount in situ hybridization of 9 hpf embryos. e1i1+P2 indicates co-injection of e1i1 morpholino with 500pg Papp-a2-encoding mRNA. Numbers indicate fractions of embryos displaying the depicted phenotype. Animal pole view, dorsal to the top.

Gene Expression Details No data available
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-pappa2 + MO4-tp53 standard conditions Prim-5 notochord undulate, abnormal
Prim-5 trunk curved ventral, abnormal
WT + MO2-pappa2 + MO4-tp53 standard conditions Prim-5 notochord undulate, abnormal
Prim-5 trunk curved ventral, abnormal
WT + MO3-pappa2 + MO4-tp53 standard conditions Prim-5 notochord undulate, abnormal
Prim-5 trunk curved ventral, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Journal of Cell Science for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ J. Cell Sci.