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A Drosophila Enhancer of split [E(spl)] bHLH protein, m delta, was misexpressed in Xenopus embryos along with green fluorescent protein (GFP) as a lineage label. The Drosophila protein translocated to the nucleus of Xenopus cells and led to neural hypertrophy in the GFP-labeled dorsal ectoderm, a phenotype similar to that caused by the misexpression of activated Xotch. Our data indicate a strong conservation in E(spl)bHLH function in the Notch signaling pathway of flies and vertebrates.
FIG. 1. (A) Whole·mount staining of a neural plate stage embryo expressing mo protein in some epidermal nuclei (arrows). Embryos were
fixed in Dent's fixative and bleached in H20 2 to remove native pigment. Detection of the antigen was with mab323 and a horseradish peroxidasecoupled
anti-mouse secondary antibody (Jackson Labs). (B) Nuclear localization of mo protein. Higher power view of stained region in A. (C)
Cement gland reduction in a living mo RNA-injected embryo. Fluorescence from GFP coexpression (within dotted lines) was used to follow the
mo expressing regions. This is a ventral view of a neural tube stage embryo (anterior up). The cement gland (cg) is the darkly pigmented tissue.
Its development has been inhibited in the GFP area. This figure, along with D and G, was taken with a Hoechst filter to show both the GFP
localization and the rest of the animal because in micrographs with the fluorescein filter alone (E), it was difficult to see anything but the GFPlabeled
areas. (D and E) Neural expansion in a living embryo caused by expression of mo RNA on the right side (anterior to left) of a tailbud
embryo. Again, fluorescence from GFP coexpression was used to follow the md expressing regions. The dotted Jines mark the peripheral boundaries
of the dorsal CNS and the midline. The eyebud region (e) could not be seen on the right. (F and G) Neural tube expansion in a late tailbud
embryo. mb and GFP were expressed on the left side. The midline is dotted, and a solid line through the hindbrain region indicates the approximate
extent of neural tube in this region. (H) Cross-section through hindbrain region in animal injected on right side with m!5 and GFP RNA.
Note the obvious expansion of the neural tube (nt) and the somitic mesoderm {s) on the mh expressing side.
