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ZFIN ID: ZDB-FIG-190624-36
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:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 14-19 somites to Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Day 4

Fig. 1

tmem33 knockdown inhibits angiogenesis and localises to the ER in ECs. adtmem33 is expressed ubiquitously during segmentation, but displays enrichment in the pronephros (black arrowheads) and somite boundaries, which is more pronounced from 24 hpf. Pronephric expression is evident in 26 hpf transverse sections (black arrows). eg Tmem33-EGFP protein localises to the nuclear envelope (blue arrowheads) and ER (white arrowheads) within the caudal artery in fli1a:DsRedEx2embryos (Scale bars 1 µm). hk tmem33 morphants injected with 0.4 ng morpholinos display delayed migration of Tg(fli1a:egfp) positive SeAs, which stall at the horizontal myoseptum (j, k, white arrowheads), compared with control Tg(fli1a:egfp) positive morphants (h, i), which begin to anastomose by 30 hpf (yellow arrowheads) (scale bars 50 µm). lo By 48 hpf, Tg(fli1a:EGFP;−0.8flt1:RFP) tmem33 morphant SeAs complete dorsal migration, but display incomplete DLAV formation (n, o, yellow arrowheads) and lack lymphatic vasculature (red arrowheads). At 48 hpf Tg(fli1a:EGFP; −0.8flt1:RFP) control morphants display secondary angiogenesis (l, m, yellow arrowheads) and parachordal lymphangioblasts (red arrowhead) (scale bars 50 µm). p tmem33 morphants injected with 0.4 ng morpholinos display reduced SeA length at 30 hpf (t-test ****p = < 0.0001; t = 4.075; DF = 24. n = 3 repeats, 10 embryos per group). q tmem33 morphants injected with 0.4 ng morpholinos display incomplete formation of DLAV (t-test ****p = < 0.0001; t = 5.618; DF = 28. n = 3 repeats, 9 or 10 embryos per group). r, s Thoracic duct formation is impaired in tmem33 morphants injected with 0.4 ng morpholinos (white asterisks), compared with control morphants (white arrowheads) (scale bars 50 µm). DA, dorsal aorta; PCV, posterior cardinal vein. Source data are provided as a Source Data file

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRedx sh495Tg standard conditions Prim-15 blood vessel endothelial cell IFL
EGFP hu5333Tg; y1Tg standard conditions Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
Long-pec posterior cardinal vein blood vessel endothelial cell IFL
Day 4 dorsal aorta blood vessel endothelial cell IFL
Day 4 intersegmental vessel blood vessel endothelial cell IFL
Day 4 posterior cardinal vein blood vessel endothelial cell IFL
hu5333Tg; y1Tg + MO1-tmem33 + MO2-tmem33 standard conditions Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental vessel blood vessel endothelial cell IFL
Long-pec posterior cardinal vein blood vessel endothelial cell IFL
Day 4 dorsal aorta blood vessel endothelial cell IFL
Day 4 intersegmental vessel blood vessel endothelial cell IFL
Day 4 posterior cardinal vein blood vessel endothelial cell IFL
y1Tg standard conditions Prim-15 dorsal aorta blood vessel endothelial cell IFL
Prim-15 intersegmental vessel blood vessel endothelial cell IFL
Prim-15 posterior cardinal vein blood vessel endothelial cell IFL
y1Tg + MO1-tmem33 + MO2-tmem33 standard conditions Prim-15 dorsal aorta blood vessel endothelial cell IFL
Prim-15 intersegmental vessel blood vessel endothelial cell IFL
Prim-15 posterior cardinal vein blood vessel endothelial cell IFL
RFP hu5333Tg; y1Tg standard conditions Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental artery blood vessel endothelial cell IFL
Day 4 dorsal aorta blood vessel endothelial cell IFL
Day 4 intersegmental artery blood vessel endothelial cell IFL
hu5333Tg; y1Tg + MO1-tmem33 + MO2-tmem33 standard conditions Long-pec dorsal aorta blood vessel endothelial cell IFL
Long-pec intersegmental artery blood vessel endothelial cell IFL
Day 4 dorsal aorta blood vessel endothelial cell IFL
Day 4 intersegmental artery blood vessel endothelial cell IFL
tmem33 WT standard conditions 14-19 somites pronephros ISH
Prim-5 pronephros ISH
Prim-5 trunk vasculature ISH
sh495Tg standard conditions Prim-5 blood vessel endothelial cell endoplasmic reticulum OTHR
Prim-5 blood vessel endothelial cell nuclear envelope OTHR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
hu5333Tg; y1Tg + MO1-tmem33 + MO2-tmem33 standard conditions Long-pec cardiovascular system lacks all parts of type vascular lymphangioblast, 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 cell migration involved in sprouting angiogenesis decreased occurrence, abnormal
Long-pec intersegmental vessel sprouting angiogenesis decreased occurrence, abnormal
Long-pec lymph vessel development decreased occurrence, abnormal
Day 4 cardiovascular system lacks all parts of type thoracic duct, abnormal
Day 4 lymph vessel development decreased occurrence, abnormal
y1Tg + MO1-tmem33 + MO2-tmem33 standard conditions Prim-15 intersegmental vessel cell migration involved in sprouting angiogenesis decreased occurrence, abnormal
Prim-15 intersegmental vessel endothelial tip cell spade-shaped, 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.