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Figure 6


Fig. 6. numb and spdo control VUM neuronal cell fate. Confocal images of stage 14-15 Drosophila embryos in sagittal view. (A-E,P) Wild-type, (F-J) numb4/numb4, (K-O) spdoG104/spdoG104, (Q) sim-Gal4 UAS-numb and (R) sim-Gal4 UAS-spdo. All embryos had sim-Gal4 UAS-tau-GFP (green) in the background, except I, which shows anti-Sim (green) staining. To identify the VUM neurons, numb mutants were double-labeled for En (not shown, except in H); spdo mutants were double-labeled with Tbh (not shown, except in L). In A-O, white arrowheads indicate cells expressing mVUM genes and yellow arrowheads indicate cells expressing iVUM genes. (A) In wild type, Zn finger homeodomain 1 (zfh1) was present in all three mVUMs and not in the iVUMs. (F) In numb, zfh1 expression was absent. (K) In spdo, zfh1 expression was expanded to five VUMs. (B) Tbh was expressed in three mVUMs in wild type. (G) In numb, Tbh was not expressed. (L) In spdo, five VUMs expressed Tbh. (C) In wild type, En was present in three iVUMs as well as other cell types, including the PMG (bracket). (H) In numb, En was present in five VUMs in addition to the PMG (bracket). (M) In spdo, En was absent from VUMs, but was present in the PMG (asterisk and bracket). (D) Glutamic acid decarboxylase 1 (Gad1) was expressed in three iVUMs in wild type. (I) Gad1 expression was expanded to six cells in numb (four of the six VUMs can be seen in this focal plane). (N) In spdo, Gad1 expression was present in only one VUM. (E) In wild type, Cas was present in two iVUMs (iVUM4,5) and two mVUMs (mVUM4,5). In (J) numb and (O) spdo mutant embryos, Cas was also present in iVUM4,5 and mVUM4,5. (P-R) Overexpression of numb causes a VUM cell fate change. (P) Wild-type expression of Tbh (magenta) in three mVUMs and of En protein (blue) in three iVUMs. (Q) In sim-Gal4 UAS-numb, six ventral Tbh+ En- mVUMs (two of the six cells are absent in this focal plane) were present. En in PMG (bracket) was unaffected. (R) sim-Gal4 UAS-spdo had a wild-type Tbh and En pattern (two of three Tbh+ mVUMs are present in this image).





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