ZFIN Image: Pan et al., 2014, Fig. 3

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Figure Caption

Fig. 3 Stable heat shock-induced overexpression of Vangl2 FL causes defects in FBM neuron migration (dominant-negative phenotype). (A) Schematic outlining the Gateway cloning strategy for generating Tol2 vectors for various Vangl2 constructs used to generate stable transgenic lines. (B) In triple transgenic 48 hpf Tg(hsp70:Gal4); Tg(UAS:Vangl2FL-nlsRFP); Tg(isl1:Gfp) experimental embryos heat shocked at 10 hpf, RFP-expressing cells (left panel, white arrowhead) were distributed broadly throughout the embryo. Myc-immunostaining (middle panel, brown) showed Vangl2 protein expressed on cell membranes (inset, white arrowhead). Importantly, anti-GFP immunostaining (right panel, brown) revealed that a majority of FBM neurons failed to migrate out of r4 (black arrowhead), while several neurons had migrated into r5 (white arrowhead). (C) By contrast, in double transgenic Tg(UAS:Vangl2FL-nlsRFP); Tg(isl1:Gfp) control embryos heat shocked at 10 hpf, RFP and Vangl2-expressing cells were absent, and FBM neurons migrated normally from r4 to r6 (white arrowhead). Asterisks in RFP panels in B and C indicate autofluorescence in the yolk tube.

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Acknowledgments

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Reprinted from Mechanisms of Development, 131, Pan, X., Sittaramane, V., Gurung, S., and Chandrasekhar, A., Structural and temporal requirements of Wnt/PCP protein Vangl2 function for convergence and extension movements and facial branchiomotor neuron migration in zebrafish, 1-14, Copyright (2014) with permission from Elsevier. Full text @ Mech. Dev.