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ZFIN ID: ZDB-FIG-180816-3
Davis et al., 2018 - ETS transcription factor Etsrp / Etv2 is required for lymphangiogenesis and directly regulates vegfr3 / flt4 expression. Developmental Biology   440(1):40-52 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Long-pec to Days 7-13
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Days 7-13

Fig. 3

Etv2 depletion by caged MO photoactivation inhibits lymphangiogenesis. (A-H) Etv2 depletion by caged MO photoactivation inhibits thoracic duct development in fli1a:GFP; kdrl:mCherry (A-D′) and TgBAC(prox1a:KalT4-UAS:uncTagRFP) (E-H) reporter zebrafish larvae at 4.5 dpf and 7 dpf, respectively. (A,E) Control uninjected larvae with normal thoracic duct (TD) below the dorsal aorta (DA). (B,F) Embryos injected with etv2 MO and caging strand solution (hereafter called “photomorph solution”), but never exposed to UV photoactivation, develop normal thoracic duct similar to uninjected controls. (C,G) Embryos injected with photomorph solution and exposed to UV photoactivation at 6 hpf (shield stage), develop abnormal vasculature (C) and no or discontinuous thoracic duct (G) compared to control embryos as expected. (D,H) Embryos injected with photomorph solution, exposed to UV photoactivation at 24 hpf, develop normal vasculature compared to control embryos but discontinuous or absent TD (asterisks). (I-L′) Etv2 is required for the formation of parachordal lymphangioblasts, as observed in fli1a:GFP; kdrl:mCherry reporter zebrafish larvae at 56 hpf. (I) Control uninjected larvae. (J) Embryos injected with photomorph solution, but never exposed to UV photoactivation, develop normal parachordal lymphangioblasts (PL, arrowheads) similar to uninjected controls. (K) Embryos injected with photomorph solution, and UV photoactivated at 6 hpf (shield stage), develop abnormal vasculature, while PLs cannot be visualized due to abnormal branching of ISVs. (L) Embryos injected with photomorph solution, and UV photoactivated at 24 hpf, develop normal vasculature compared to control embryos while PLs are absent (arrowheads). A′-D′ and I′-L′ show magnified areas indicated by boxes in A-D and I-L. Each experiment has been repeated at least three times. Number of larvae showing a displayed phenotype out of the total number of larvae is shown in the upper right corner.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP ci5Tg; y1Tg control Long-pec trunk vasculature IFL
Long-pec vascular lymphangioblast IFL
Day 4 thoracic duct IFL
Day 4 trunk vasculature IFL
ci5Tg; y1Tg standard conditions Long-pec trunk vasculature IFL
Long-pec vascular lymphangioblast IFL
Day 4 thoracic duct IFL
Day 4 trunk vasculature IFL
ci5Tg; y1Tg + MO2-etv2 standard conditions Long-pec trunk vasculature IFL
Long-pec Not Detected vascular lymphangioblast IFL
Day 4 Not Detected thoracic duct IFL
Day 4 trunk vasculature IFL
y1Tg ; nim5Tg control Days 7-13 dorsal aorta IFL
Days 7-13 trunk vasculature IFL
y1Tg ; nim5Tg standard conditions Days 7-13 dorsal aorta IFL
Days 7-13 trunk vasculature IFL
y1Tg ; nim5Tg + MO2-etv2 standard conditions Days 7-13 dorsal aorta IFL
Days 7-13 trunk vasculature IFL
mCherry ci5Tg; y1Tg control Long-pec lymph vasculature IFL
Day 4 trunk vasculature IFL
ci5Tg; y1Tg standard conditions Long-pec lymph vasculature IFL
Day 4 trunk vasculature IFL
ci5Tg; y1Tg + MO2-etv2 standard conditions Long-pec lymph vasculature IFL
Day 4 trunk vasculature IFL
TagRFP y1Tg ; nim5Tg control Days 7-13 lymph vasculature IFL
Days 7-13 thoracic duct IFL
y1Tg ; nim5Tg standard conditions Days 7-13 lymph vasculature IFL
Days 7-13 thoracic duct IFL
y1Tg ; nim5Tg + MO2-etv2 standard conditions Days 7-13 Not Detected thoracic duct IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ci5Tg; y1Tg + MO2-etv2 standard conditions Long-pec lymphangiogenesis decreased occurrence, abnormal
Long-pec trunk vasculature morphology, abnormal
Long-pec vascular lymphangioblast EGFP expression absent, abnormal
Long-pec vascular lymphangioblast absent, abnormal
Day 4 lymphangiogenesis decreased occurrence, abnormal
Day 4 thoracic duct absent, abnormal
Day 4 thoracic duct EGFP expression absent, abnormal
Day 4 trunk vasculature morphology, abnormal
y1Tg ; nim5Tg + MO2-etv2 control Days 7-13 thoracic duct absent, abnormal
Days 7-13 trunk vasculature morphology, abnormal
y1Tg ; nim5Tg + MO2-etv2 standard conditions Days 7-13 thoracic duct TagRFP expression absent, abnormal
Days 7-13 thoracic duct absent, abnormal
Days 7-13 trunk vasculature morphology, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 440(1), Davis, J.A., Koenig, A.L., Lubert, A., Chestnut, B., Liu, F., Desai, S.P., Winkler, T., Pociute, K., Choi, K., Sumanas, S., ETS transcription factor Etsrp / Etv2 is required for lymphangiogenesis and directly regulates vegfr3 / flt4 expression, 40-52, Copyright (2018) with permission from Elsevier. Full text @ Dev. Biol.