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ZFIN ID: ZDB-FIG-110527-4
Krueger et al., 2011 - Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo. Development (Cambridge, England)   138(10):2111-2120 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-15
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-15

Fig. 4 Tip and stalk cell marker expression and Notch signaling in flt1 morphants. (A,B) Expression of kdrl, flt4 and dll4 in control and flt1 morphants. Expression of the tip cell marker kdrl in segmental sprouts in Tg(kdrl:hras-mcherry)s896 transgenic zebrafish embryos (A, left) and kdrl in toto in situ hybridization (A, overview and detail). Note the ectopic expression of flt4 in the dorsal aorta of flt1 morphants (A, fourth panel). Expression of the stalk cell markers nrarpa and nrarpb is reduced, but expression of dll4 is maintained in morphants (A, fifth panel; B). (C,D) In situ hybridization and Taqman analysis of the trunk region show reduced expression of the receptors notch1a, notch1b, notch2 and notch3 and of the Notch ligands jag1a, jag1b and jag2 in flt1 morphants. Reduced expression of efnb2a suggests loss of Notch signaling in flt1 morphants. (E) Conditional overexpression of notch1a intracellular cleaved domain (NICD) in flt1 morphants (right) rescues segmental arterial branching defects. (F) Segmental branching pattern in dll4 morphants (red arrowhead) or in embryos treated with the Notch γ-secretase inhibitor DAPT (red arrow) did not phenocopy the branching pattern of flt1 morphants (yellow arrowhead). The boxed regions are shown at higher magnification in the right-hand panels. DA, dorsal aorta; PCV, posterior cardinal vein. *, P<0.05; **, P<0.01; Student′s t-test. Error bars indicate s.e.m. f.c., fold change relative to age-matched control embryos.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
dll4 WT control Prim-15 dorsal aorta ISH
Prim-15 intersegmental vessel ISH
Prim-15 whole organism RTPCR
WT + MO3-flt1 standard conditions Prim-15 dorsal aorta ISH
Prim-15 intersegmental vessel ISH
Prim-15 whole organism RTPCR
flt4 WT control Prim-15 posterior cardinal vein ISH
WT + MO3-flt1 standard conditions Prim-15 dorsal aorta ISH
Prim-15 posterior cardinal vein ISH
kdrl WT control Prim-15 dorsal aorta ISH
Prim-15 intersegmental vessel endothelial tip cell ISH
WT + MO3-flt1 standard conditions Prim-15 dorsal aorta ISH
Prim-15 intersegmental vessel endothelial tip cell ISH
mCherry s896Tg control Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
s896Tg + MO3-flt1 standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
nrarpa WT control Prim-15 whole organism RTPCR
WT + MO3-flt1 standard conditions Prim-15 whole organism RTPCR
nrarpb WT control Prim-15 whole organism RTPCR
WT + MO3-flt1 standard conditions Prim-15 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s896Tg + MO3-flt1 standard conditions Prim-15 intersegmental vessel endothelial tip cell increased width, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development