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ZFIN ID: ZDB-FIG-110527-5
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:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-15 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. 5 Distribution of Flt1 in vessels and nerves. (A) Comparison of flt1BAC:yfp and kdrl:cherry expression in segmental artery sprouts of 30 hpf Tg(kdrl:ras-cherry)s916 × Tg(flt1BAC:yfp) double-transgenic zebrafish embryos, in controls (top row) and flt1 morphants (bottom row). In controls, note the overlap of flt1BAC:yfp and kdrl:cherry expression in tip and stalk cells. At 30 hpf, expression of flt1BAC:yfp is restricted to the vascular compartment. After injection of flt1 ATG-blocking MO, expression of flt1BAC:yfp in sprouts appears less intense, but is not completely lost. (B) Injection of flt1 ATG-blocking MO in Tg(kdrl:ras-cherry)s916 × Tg(flt1BAC:yfp) double-transgenic embryos causes hyperbranching of segmental vessels and affects neurons. In controls at 48 hpf, flt1BAC:yfp marks segmental vessels and a subset of spinal cord neurons (arrowhead). Injection of the flt1 ATG-blocking MO induces aberrant vessel patterning and the neuronal flt1BAC:yfp expression domain is reduced. Injection of the flt1 ATG-blocking MO in Tg(kdrl:hras-mcherry)s896 × Tg(huC:egfp) double-transgenic embryos affects neurons. (C-E3) Flt1 immunostaining in Tg(huC:egfp) neuronal reporter embryos shows Flt1 throughout the neural tube. Overview of neural tube (C-C3) and different focal planes from dorsal to ventral (D-E3) are shown. (F) Flt1 immunostaining in Tg(flt1BAC:yfp) embryos shows co-localization of flt1BAC:yfp with Flt1 antibody staining in spinal cord neurons (boxed). (G) Immunostaining for Flt1 and zn-12 shows Flt1 staining of Rohon-Beard sensory neurons. (H) Overexpression of fli1ep:cherry-sflt1 in Tg(huC:egfp) embryos results in neuronal sFlt1-cherry expression throughout the neural tube. Arrowheads indicate co-localization of huC:egfp with sFlt1-cherry on neuronal cell bodies. (I) Overexpression of kdrl:egfp-sflt1 in Tg(kdrl:hras-mcherry)s896 embryos shows sFlt1-egfp expression in vessels and the neural tube (arrowheads). Nt, neural tube; Se, segmental artery.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP knu3Tg standard conditions Prim-15 spinal cord motor neuron IFL
Prim-15 spinal cord sensory neuron IFL
Prim-15 spinal cord interneuron IFL
knu3Tg; s896Tg standard conditions Long-pec spinal cord neuron IFL
knu3Tg; s896Tg + MO3-flt1 standard conditions Long-pec spinal cord neuron IFL
y1Tg standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel IFL
Prim-15 Rohon-Beard neuron IFL
flt1 hu4624Tg standard conditions Prim-25 spinal cord neuron IHC
knu3Tg standard conditions Prim-15 spinal cord motor neuron IHC
Prim-15 spinal cord sensory neuron IHC
Prim-15 spinal cord interneuron IHC
mCherry hu4624Tg; s916Tg standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
Long-pec dorsal aorta IFL
hu4624Tg; s916Tg + MO3-flt1 standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
Long-pec dorsal aorta IFL
knu3Tg; s896Tg standard conditions Long-pec dorsal aorta IFL
Long-pec dorsal longitudinal anastomotic vessel IFL
Long-pec intersegmental vessel IFL
knu3Tg; s896Tg + MO3-flt1 standard conditions Long-pec dorsal aorta IFL
Long-pec dorsal longitudinal anastomotic vessel IFL
Long-pec intersegmental vessel IFL
YFP hu4624Tg standard conditions Prim-25 dorsal longitudinal anastomotic vessel IFL
Prim-25 intersegmental vessel IFL
Prim-25 spinal cord neuron IFL
hu4624Tg; s916Tg standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
Long-pec intersegmental vessel IFL
Long-pec spinal cord neuron IFL
hu4624Tg; s916Tg + MO3-flt1 standard conditions Prim-15 dorsal aorta IFL
Prim-15 intersegmental vessel IFL
Prim-15 intersegmental vessel endothelial tip cell IFL
Long-pec intersegmental vessel IFL
Long-pec spinal cord neuron IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
zn-12 IHC y1Tg standard conditions Prim-15 Rohon-Beard neuron
Phenotype Details
Fish Conditions Stage Phenotype
hu4624Tg; s916Tg + MO3-flt1 standard conditions Long-pec intersegmental artery branched, abnormal
knu3Tg; s896Tg + MO3-flt1 standard conditions Long-pec spinal cord has fewer parts of type CNS neuron (sensu Vertebrata), 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