ZFIN ID: ZDB-FIG-170510-5
Kasper et al., 2017 - MicroRNAs Establish Uniform Traits during the Architecture of Vertebrate Embryos. Developmental Cell   40:552-565.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Conditions:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 6

miR-139 and miR-24 Mutants Are Broadly Affected by Stress

(A and B) Schematic depicts the duration of chemical exposure or environmental condition used to test stress sensitivity of miR-139-regulated (A) and miR-24-regulated (B) vascular traits in the absence of miRNA activity. Stressors were administered at doses that minimally affect wild-type as indicated (see Table S1).

(C and D) Arrows point to ectopic ISV branches in 54-hpf miR-139Δ/Δ embryos treated with pro-angiogenic drugs: BIO (n = 24) and GSI (n = 50–55 embryos). Controls were treated with DMSO vehicle (n = 17–24 embryos).

(E) Arrows highlight stunted ISV branches in 27-hpf miR-139Δ/Δ embryos treated with anti-angiogenic VEGF inhibitor SU5416 (n = 25–27 embryos). Controls were treated with DMSO vehicle (n = 17–25 embryos).

(F) Arrows show ectopic ISV branches in 54-hpf miR-139Δ/Δ embryos exposed to high-temperature stress (n = 31–44 embryos/genotype).

(G) Yellow arrows show region captured in zoom images. White arrow points to hypersprouting of DLAV endothelial cells in 52 hpf miR-139Δ/Δ embryos exposed to 4 hr of hypoxia (n = 40–45 embryos/genotype).

(H) Arrows indicate the HA of BIO-treated embryos.

(I–M) Quantification of 56-hpf HA length (μm) upon treatment of BIO or DMSO (n = 12–24 for all genotypes except n = 6 for quadruple mutant embryos, I); GSI or DMSO (n = 15, J); SU5416 or DMSO (n = 11–16, K); high or control temperature (n = 38–50, L); and hypoxia or normoxia (n = 24, M). S = single, D = double, T = triple, Q = quadruple mutant embryos.

All images were captured with a 25× objective. All bar plots represent mean ± SEM, and significance comparisons between samples are as indicated. n.s., not significant (p > 0.05); p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001, two-tailed Mann-Whitney U test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP la116Tg chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg chemical treatment by environment: EC 3.4.23.46 (memapsin 2) inhibitor Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg chemical treatment by environment: semaxanib Prim-5 dorsal aorta IHC
Prim-5 intersegmental vessel IHC
Prim-5 posterior cardinal vein IHC
Prim-5 trunk vasculature IHC
la116Tg heat exposure Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg hypoxia Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg ; mir139ya302/ya302 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg ; mir139ya302/ya302 chemical treatment by environment: EC 3.4.23.46 (memapsin 2) inhibitor Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg ; mir139ya302/ya302 chemical treatment by environment: semaxanib Prim-5 dorsal aorta IHC
Prim-5 intersegmental vessel IHC
Prim-5 posterior cardinal vein IHC
Prim-5 trunk vasculature IHC
la116Tg ; mir139ya302/ya302 heat exposure Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg ; mir139ya302/ya302 hypoxia Long-pec dorsal aorta IHC
Long-pec intersegmental vessel IHC
Long-pec posterior cardinal vein IHC
Long-pec trunk vasculature IHC
la116Tg ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-1ya324/ya324 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-1ya324/ya324 ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-4ya327/ya327 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
la116Tg ; mir24-4ya327/ya327 ; mir24-3ya326/ya326 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-1ya324/ya324 ; la116Tg ; mir24-2ya325/ya325 ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-1ya324/ya324 ; la116Tg ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-2ya325/ya325 ; la116Tg ; mir24-3ya326/ya326 ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-2ya325/ya325 ; mir24-1ya324/ya324 ; mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-3ya326/ya326 ; la116Tg chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec cranial vasculature IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
la116Tg ; mir139ya302/ya302 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec intersegmental vessel increased branchiness, abnormal
la116Tg ; mir139ya302/ya302 chemical treatment by environment: EC 3.4.23.46 (memapsin 2) inhibitor Long-pec intersegmental vessel increased branchiness, abnormal
la116Tg ; mir139ya302/ya302 chemical treatment by environment: semaxanib Prim-5 intersegmental vessel decreased length, abnormal
la116Tg ; mir139ya302/ya302 heat exposure Long-pec intersegmental vessel increased branchiness, abnormal
la116Tg ; mir139ya302/ya302 hypoxia Long-pec dorsal longitudinal anastomotic vessel increased branchiness, abnormal
la116Tg ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-1ya324/ya324 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-1ya324/ya324 ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-4ya327/ya327 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
la116Tg ; mir24-4ya327/ya327 ; mir24-3ya326/ya326 ; mir24-2ya325/ya325 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-1ya324/ya324 ; la116Tg ; mir24-2ya325/ya325 ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-1ya324/ya324 ; la116Tg ; mir24-4ya327/ya327 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-2ya325/ya325 ; la116Tg ; mir24-3ya326/ya326 ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-2ya325/ya325 ; mir24-1ya324/ya324 ; mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-2ya325/ya325 ; mir24-1ya324/ya324 ; mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: EC 3.4.23.46 (memapsin 2) inhibitor Long-pec hypobranchial artery decreased length, abnormal
mir24-2ya325/ya325 ; mir24-1ya324/ya324 ; mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: semaxanib Long-pec hypobranchial artery decreased length, abnormal
mir24-2ya325/ya325 ; mir24-1ya324/ya324 ; mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 hypoxia Long-pec hypobranchial artery decreased length, abnormal
mir24-3ya326/ya326 ; la116Tg chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
mir24-4ya327/ya327 ; la116Tg ; mir24-3ya326/ya326 ; mir24-1ya324/ya324 chemical treatment by environment: 6-bromoindirubin-3'-oxime Long-pec hypobranchial artery decreased length, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 40, Kasper, D.M., Moro, A., Ristori, E., Narayanan, A., Hill-Teran, G., Fleming, E., Moreno-Mateos, M., Vejnar, C.E., Zhang, J., Lee, D., Gu, M., Gerstein, M., Giraldez, A., Nicoli, S., MicroRNAs Establish Uniform Traits during the Architecture of Vertebrate Embryos, 552-565.e5, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell