Figure 5

Mutations affecting neuroblast proliferation and chromosome separation. (A-D) Mutations affecting cell proliferation. DNA staining (blue); Miranda (red); CD8::GFP (green); Bar: 10 μm. Wild type clones (A) contain a single large neuroblast and a number of smaller progeny. B18 clones (B) contain fewer progeny cells and we rarely observe mitotic neuroblasts. In clones of E45, which is allelic to makos, we also see few progeny but neuroblasts have strong crescents of Miranda and appear arrested in a metaphase-like state, although some separation of sister chromatids is observed (C). (D-I) Mutations affecting chromosome separation. DNA staining (blue); Miranda (red, D-H'); CD8::GFP (green, D, E'); Inscuteable (green, F, F", I, I'); Centrosomin (green, H, H'); γ-tubulin (red, I, I'); Bar: 10 μm. Both DL42 (D) and C93 (E, E') clones contain large cells with a high DNA content which never appear to divide. C93 is allelic to separase, and C93/Df(3L)Exel6106 hemizygote (F-F'') or C93/sse^M13transheterozygote (G) neuroblasts are large with a high DNA content. In hemizygous neuroblasts we observe multiple crescents of Miranda (F, F') and Inscuteable (F, F''), which never overlap. Labelling with anti-Centrosomin (H, H') or γ-tubulin (I, I') reveals that C93 hemizygous neuroblasts contain multiple centrosomes.