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First published online 11 October 2006
doi: 10.1242/dev.02643

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Uncoupling Dorsal-mediated activation from Dorsal-mediated repression in the Drosophila embryo

Girish S. Ratnaparkhi, Songtao Jia^* and Albert J. Courey

Department of Chemistry and Biochemistry, University of California at Los Angeles, Los Angeles, CA 90095-1569, USA.

View larger version (20K): [in a new window]   Fig. 1. Activation and repression by DL variants in S2 cells. (A) Structure of DL variants. (B) Activation of the D4-luciferase reporter by addition of increasing amounts (50 ng, 150 ng) of vectors encoding DL variants. (Inset) Anti-DL immunoblot of lysates of S2 cells transiently expressing the FLAG-DL constructs. (C) Synergistic activation of the DE5-luciferase reporter by vectors encoding DL variants (60 ng) and TWI (20 ng). An expression vector encoding GRO (500 ng) was also included in the indicated transfection mixtures. (D) Synergistic activation of the G5DE5-luciferase reporter by vectors encoding DL variants (60 ng) and TWI (20 ng). gro dsRNA (1 µg) was also included in the indicated transfection mixtures. (Inset) Anti-GRO immunoblot of lysates of S2 cells treated or not treated with gro dsRNA.

View larger version (46K): [in a new window]   Fig. 2. Activation and repression of DL target genes by maternally expressed DL variants. (A-F) The expression of target genes was monitored by in situ hybridization using digoxigenin-labeled probes against twi, sna, sog or brk (genes activated by DL), and dpp or zen (genes repressed by DL). Sagittal views of the embryos are oriented with anterior on the left and the dorsal side at the top. Maternal genotypes are described in the Materials and methods.

View larger version (39K): [in a new window]   Fig. 3. Activation and repression by DL variants visualized by double labeling. (A-D) TWI protein expression was monitored by antibody staining (red, left), while zen mRNA expression was monitored by in situ hybridization (blue, right). Sagittal, confocal sections of the embryos are oriented with anterior on the left and the dorsal side at the top. Maternal genotypes are described in the Materials and methods.

View larger version (48K): [in a new window]   Fig. 4. DV patterning of the cuticle is rescued by dedicated activator DL variants. (A-H) Cuticle preparations of fully developed embryos expressing DL variants produced by females of the indicated genotypes (see Materials and methods). Anterior is to the left. (I) DPP/BMP signaling is redundantly blocked during DV patterning in invertebrate and vertebrate embryos.