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ZFIN ID: ZDB-FIG-170220-33
Wild et al., 2017 - Neuronal sFlt1 and Vegfaa determine venous sprouting and spinal cord vascularization. Nature communications   8:13991 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Pec-fin to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage Range: Pec-fin to Day 4

Fig. 3

Imaging and quantification of sprouting kinetics in flt1ka601 mutants.

(a) Time lapse imaging of sprout initiation and anastomosis formation in flt1ka601 mutant. Sprout initiation (60.0 hpf), elongation (65.2 hpf) and connection-anastomoses (72.5 hpf) with adjacent aISV. (b) Time lapse imaging of sprout initiation and retraction in flt1ka601 mutant. Endothelial cells produce filopodia (68.8 hpf), extend a sprout (71.6 hpf), which subsequently retracts (80.8 hpf). (c) Quantification of data in (a,b) showing % of sprouts retracting (top part, 45%) or connecting (bottom part, 55%) to adjacent ISVs. In the latter scenario sprouts in 95% of cases connected to aISV (red) and in 5% of cases to vISV (blue). Angiogenic behaviour was analysed in time-lapse confocal movies, n=20 embryos. (df) Filopodia directionality and length in flt1ka601 mutants (n=10 embryos, n=920 filopodia). (gj) Time lapse imaging of endothelial nuclei in Tg(fli1a:nGFP)y7; Tg(kdrl:hsa.HRAS-mcherry)s916 showing association between nuclear position and sprouting initiation point (SIP). Note that sprouts arise in close proximity to the position of the nucleus. Arrowheads indicate sprouts; nuclei at indicated time points (sprout initiation with actively migrating nucleus towards SIP I, II, III, IV and nucleus already located at SIP 1,2,3,4). (k) Schematic representation of nuclear position with respect to SIP. (l,m) Quantification of observations in gj. Red dot indicates sprout initiation time point. Note that sprouting preferentially occurs when endothelial nuclei are within less than 5 μm from SIP (SIP below dotted line in (l)). n=5 (l) and n=13 (m). (nq) Quantification of EC nuclei in aISV, vISVs, DLAV, DA and PCV of WT and flt1ka601 embryos at 4 dpf; mean±s.e.m., t-test, n=21 embryos per genotype. (r) Quantification of DLAV width in WT and flt1ka601 mutant, n=9 embryos per genotype. A, artery; aISV, arterial intersegmental vessel; DA, dorsal aorta; DLAV, dorsal longitudinal anastomotic vessel; EC, endothelial cell; PCV, posterior cardinal vein; SIP, sprout initiation point; V, vein; vISV, venous intersegmental vessel. Scale bar, 30 μm in a,b; 10 μm in d,e,gj; 50 μm in o,p.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP s843Tg ; flt1ka601/ka601 standard conditions Pec-fin to Protruding-mouth intersegmental artery IFL
Pec-fin to Protruding-mouth intersegmental vein vascular sprouts IFL
y7Tg standard conditions Day 4 trunk vasculature nucleus IFL
y7Tg ; flt1ka601/ka601 standard conditions Day 4 trunk vasculature nucleus IFL
y7Tg ; s916Tg ; flt1ka601/ka601 standard conditions Pec-fin dorsal longitudinal anastomotic vessel nucleus IFL
Pec-fin intersegmental vein vascular sprouts IFL
Pec-fin intersegmental vessel nucleus IFL
Pec-fin vascular sprouts nucleus IFL
mCherry y7Tg ; s916Tg ; flt1ka601/ka601 standard conditions Pec-fin dorsal longitudinal anastomotic vessel IFL
Pec-fin intersegmental vein IFL
Pec-fin intersegmental vein vascular sprouts IFL
Pec-fin intersegmental vessel IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s843Tg ; flt1ka601/ka601 standard conditions Pec-fin - Protruding-mouth intersegmental vein vascular sprouts increased amount, abnormal
y7Tg ; flt1ka601/ka601 standard conditions Day 4 dorsal aorta endothelial cell increased amount, abnormal
Day 4 dorsal longitudinal anastomotic vessel width, normal
Day 4 dorsal longitudinal anastomotic vessel endothelial cell increased amount, abnormal
Day 4 intersegmental artery endothelial cell increased amount, abnormal
Day 4 intersegmental vein endothelial cell increased amount, abnormal
Day 4 posterior cardinal vein endothelial cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Nature communications for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Nat. Commun.