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ZFIN ID: ZDB-FIG-190624-44
Savage et al., 2019 - tmem33 is essential for VEGF-mediated endothelial calcium oscillations and angiogenesis. Nature communications   10:732 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 7

SOCE inhibition limits sprouting angiogenesis and disrupts signalling pathways downstream of VEGF. a, b SKF9635 impairs angiogenesis in Tg(fli1a:EGFP) embryos (white arrowheads) and induces abnormal tip cell morphology (blue arrowhead) by 27 hpf. Scale bars 50 µm. c SKF-96365-treated embryos display reduced SeA length by 27 hpf (unpaired t-test; p = < 0.0001; t = 11.22 DF = 14; 2 repeats, n = 4 embryos per group). d, e SKF-96365 treatment impairs DLAV formation (white asterisks) in comparison with control embryos, which form the DLAV normally by 50 hpf (white arrowheads). Abnormal tip cell morphology (blue arrowhead) remain in SKF-96365-treated embryos by 50 hpf. Scale bars 50 µm. f SKF-96365-treated embryos display increased frequency of abnormal vascular branching (stunted SeAs or incomplete DLAV anastomosis) by 50 hpf (unpaired t-test; p = < 0.0001; t = 14.28 DF = 18. n = 2 repeats, n = 5 embryos per group). g tmem33 expression is not significantly altered by SKF-96365 treatment (unpaired t-test; t = 0.911, DF = 4, n = 3 repeats). hj SKF-96365 reduces frequency of endothelial Ca2+ oscillations in Tg(fli1a:gal4FFubs3;uas-gcamp7a) embryos. Intensity projection over time showing calcium oscillations in control embryos, highlighting intensity change over duration of time lapse (unpaired t-test; p = < 0.05; t = 2.594, DF = 10, n = 2 repeats, 3 embryos per group). k, l Notch reporter expression is reduced in SeAs (white arrowheads) and DA (red arrowhead) of Tg(csl:venus) embryos. Scale bars 50 µm. m DA expression of Notch reporter is reduced by SKF treatment (unpaired t-test, **p = 0.0097, t = 3.982 DF = 15, 2 repeats, n = 4 embryos per group). n, o SKF-96365 treatment reduces endothelial filopodia number (black arrowheads) and abnormal spade-shaped tip cell morphology (blue arrowhead), in Tg(fli1a:lifeact-mClover) embryos. Scale bars 5 µm. p SKF-96365 treatment reduces mean filopodia number per SeA per embryo by 26 hpf (unpaired t-test; p = < 0.05; t = 2.525, DF = 10, n = 2 repeats, n = 5 embryos per group). Source data are provided as a Source Data file

 

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP y1Tg control Prim-5 dorsal aorta blood vessel endothelial cell IFL
Prim-5 intersegmental vessel blood vessel endothelial cell IFL
Prim-5 intersegmental vessel endothelial tip cell IFL
Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec dorsal longitudinal anastomotic vessel blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
y1Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 dorsal aorta blood vessel endothelial cell IFL
Prim-5 intersegmental vessel blood vessel endothelial cell IFL
Prim-5 intersegmental vessel endothelial tip cell IFL
Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec dorsal longitudinal anastomotic vessel blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
GCaMP ubs3Tg ; sh392Tg control Prim-5 intersegmental vessel endothelial tip cell IFL
ubs3Tg ; sh392Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 intersegmental vessel endothelial tip cell IFL
mClover sh467Tg control Prim-5 endothelial tip cell filamentous actin IFL
Prim-5 intersegmental vessel endothelial tip cell IFL
sh467Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 endothelial tip cell filamentous actin IFL
Prim-5 intersegmental vessel endothelial tip cell IFL
tmem33 WT control Prim-5 whole organism RTPCR
WT chemical treatment by environment: SKF-96365 hydrochloride Prim-5 whole organism RTPCR
Venus qmc61Tg control Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
qmc61Tg chemical treatment by environment: SKF-96365 hydrochloride Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
qmc61Tg chemical treatment by environment: SKF-96365 hydrochloride Long-pec dorsal aorta blood vessel endothelial cell Venus expression decreased amount, abnormal
Long-pec dorsal aorta Notch signaling pathway decreased occurrence, abnormal
Long-pec intersegmental vessel blood vessel endothelial cell Venus expression decreased amount, abnormal
sh467Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 intersegmental vessel blood vessel endothelial cell spade-shaped, abnormal
Prim-5 intersegmental vessel endothelial tip cell has fewer parts of type endothelial tip cell filopodium, abnormal
Prim-5 intersegmental vessel endothelial tip cell filopodium assembly decreased occurrence, abnormal
ubs3Tg ; sh392Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 endothelial tip cell calcium-mediated signaling decreased occurrence, abnormal
Prim-5 endothelial tip cell regulation of cytosolic calcium ion concentration decreased occurrence, abnormal
Prim-5 endothelial tip cell regulation of store-operated calcium channel activity decreased occurrence, abnormal
Prim-5 intersegmental vessel endothelial tip cell GCaMP expression decreased amount, abnormal
y1Tg chemical treatment by environment: SKF-96365 hydrochloride Prim-5 intersegmental vessel decreased length, abnormal
Prim-5 intersegmental vessel blood vessel endothelial cell spade-shaped, abnormal
Prim-5 intersegmental vessel sprouting angiogenesis decreased occurrence, abnormal
Long-pec dorsal longitudinal anastomotic vessel malformed, abnormal
Long-pec dorsal longitudinal anastomotic vessel split, abnormal
Long-pec dorsal longitudinal anastomotic vessel sprouting angiogenesis decreased occurrence, abnormal
Long-pec intersegmental vessel decreased length, 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.