ZFIN ID: ZDB-FIG-101104-1
Lanahan et al., 2010 - VEGF Receptor 2 Endocytic Trafficking Regulates Arterial Morphogenesis. Developmental Cell   18(5):713-724 Full text @ Dev. Cell
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-25
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: 14-19 somites to Prim-25

Fig. 5  Myo6aKD Impairs Arterial Development in Zebrafish; Functional Effects of Mysosin VI Gene Disruption in Mice

(A and B) Transverse section through the trunk of a 28 hpf zebrafish embryo stained by whole-mount in situ hybridization using a myo6a specific antisense (A) or sense (B) riboprobe. Myo6a expression was observed in the neural tube (NT), somites (S), and both the dorsal aorta (black arrow) and posterior cardinal vein (white arrow). The observed expression pattern is specific, as no signal was observed upon hybridization using the sense probe.

(C and D) Dorsal view of 16 hpf Tg(fli1:EGFP)y1 embryo, head on top. In control embryos (C), GFP+ angioblasts migrated in an ordered anteroposteriorly directed zipper-like pattern from the lateral plate mesoderm toward the midline, where they assembled into the primitive axial vessels. In contrast, in myo6aKD embryos (D), a portion of angioblasts (arrows) stalled along their lateromedial movement and others failed to maintain their correct stereotyped trajectory, resulting in apparently chaotic migration pattern.

(E and F) Transverse sections through the trunk of Tg(fli1:EGFP)y1embryos of 30 hpf, following a whole-mount immunostaining using an anti-GFP antibody (green) and counterstained with DAPI (blue); head on top. Compared to control embryos (E and E′), myo6aKD embryos had a strikingly thinner dorsal aorta (arrow) with an obvious reduced lumen size, while the posterior cardinal vein (arrowhead) remained unaffected (F and F′). (E′) and (F′) are magnifications of the axial vessels in (E) and (F), respectively.

(G and H) Lateral view on a trunk segment of Tg(fli1:EGFP)y1embryos at 40 hpf; head to the left. In control embryos (G), ISVs sprouted bilaterally from the dorsal aorta adjacent to the ventral somite boundaries and navigated upwards to the laterodorsal roof of the neural tube, where they split, elongated, and fused to form the DLAV. However, in myo6aKD embryos (H), ISVs often consisted of slender endothelial cells (arrowheads) and/or stalled along their dorsal trajectory (arrows), thereby impairing proper DLAV formation (asteriks).

Scale bars represent 10 μm in (A), (B), and (E–F′) and 5 μm in (C), (D), (G), (H). See also Figures S3 and S4.

(I and J) Representative reconstructed micro-CT images of whole kidneys (16 μm resolution; n = 3) from age- and gender-matched (I) myosinVI+/+ and (J) myosinVI-/- mice. Note marked reduction in branching in myosinVI-/- mice.

(K) Quantitative analysis of micro-CT images indicates a marked decrease in total number of <100 µm diameter vessels in myosinVI-/- mice (white bars) relative to myosinVI+/+ mice (black bars). (Mean ± SEM, *p < 0.05).

(L) Quantitative analysis of laser Doppler images indicates significant alterations in hindlimb reperfusion 14 days after femoral artery ligation in myosinVI-/- mice (white bars) relative to myosinVI+/+ mice (black bars). (Mean ± SEM, *p < 0.05).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP y1Tg standard conditions 14-19 somites angioblastic mesenchymal cell IFL
14-19 somites axial blood vessel IFL
Prim-15 dorsal aorta IHC
Prim-15 posterior cardinal vein IHC
Prim-25 dorsal aorta IFL
Prim-25 dorsal longitudinal anastomotic vessel IFL
Prim-25 intersegmental vessel IFL
y1Tg + MO1-myo6a standard conditions 14-19 somites angioblastic mesenchymal cell IFL
14-19 somites axial blood vessel IFL
Prim-15 dorsal aorta IHC
Prim-15 posterior cardinal vein IHC
Prim-25 dorsal aorta IFL
Prim-25 dorsal longitudinal anastomotic vessel IFL
Prim-25 intersegmental vessel IFL
myo6a WT standard conditions Prim-5 dorsal aorta ISH
Prim-5 neural tube ISH
Prim-5 posterior cardinal vein ISH
Prim-5 somite ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
y1Tg + MO1-myo6a standard conditions 14-19 somites angioblast cell migration from lateral mesoderm to midline disrupted, abnormal
14-19 somites angioblastic mesenchymal cell mislocalised, abnormal
14-19 somites artery development disrupted, abnormal
14-19 somites axial blood vessel malformed, abnormal
Prim-15 dorsal aorta decreased diameter, abnormal
Prim-25 artery development disrupted, abnormal
Prim-25 axial blood vessel malformed, abnormal
Prim-25 dorsal aorta decreased diameter, abnormal
Prim-25 dorsal longitudinal anastomotic vessel malformed, abnormal
Prim-25 intersegmental vessel blood vessel endothelial cell slender, 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, 18(5), Lanahan, A.A., Hermans, K., Claes, F., Kerley-Hamilton, J.S., Zhuang, Z.W., Giordano, F.J., Carmeliet, P., and Simons, M., VEGF Receptor 2 Endocytic Trafficking Regulates Arterial Morphogenesis, 713-724, Copyright (2010) with permission from Elsevier. Full text @ Dev. Cell