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Fig. 10. Localization of JMD-deleted E-cadherin and E-catenin-EGFP in fixed and living embryos. (A) Immunohistological examination of mE-cadhJMD expression in transverse sections of stage 21 embryos. Embryos were injected at the 16-cell stage and fixed for sectioning at stage 21. After sectioning, immunohistochemical staining was performed to localize mE-cadherinJMD proteins, and sections were examined with a fluorescence (a-c) or a laser scanner confocal microscope (d-f). Embryos injected with mE-cadhJMD (500 pg) alone show a high and uniform concentration of mE-cadherinJMD at the cell-cell contacts in the cranial NCCs (a,d). Expression of mE-cadherinJMD in embryos co-injected with dominant negative (DN) Rac (50 pg) is more scattered (b,e) while an overall reduced staining is observed upon co-injection of 150 pg wild type (WT) RhoA RNA (c,f). All sections are oriented with dorsal side up. Staining in the ectoderm is indicated by an arrow, and the boxed area in the migrating neural crest cells is examined by confocal microscopy. Scale bar, 20 µm. (B) Confocal in vivo analysis of the ectodermal cell layer in tailbud-stage embryos injected at the 16-cell stage in the ventral-animal region with a plasmid encoding E-catenin-EGFP alone (a) or with the plasmid in combination with 500 pg mE-cadhJMD RNA (b) and additionally 50 pg DN Rac (c) or 150 pg WT RhoA RNA (d). While injection of mE-cadherinJMD induces strong localization of E-catenin-EGFP at the cell-cell contacts, this is greatly reduced upon co-injection of DN Rac but not by WT RhoA. Scale bars, 20 µm.

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