Microarray comparison of anterior and posterior Drosophila wing imaginal disc cells

ABSTRACT: Signaling between cells in the Anterior (A) and Posterior (P) compartments directs Drosophila wing disc development and is dependent on expression of the homeodomain transcription factor Engrailed (En) in P cells. Downstream of en, posteriorly expressed Hedgehog (Hh) protein signals across the A/P border to establish a developmental organizer that directs pattern formation and growth throughout the wing primordium. Here we extend investigations of the processes downstream of en by using expression array analysis to compare A and P cells. 102 candidate genes were identified that express differentially in the A and P compartments; four were characterized: Stubble (Sb) expression is restricted to A cells due to repression by en. CG15905, CG16884, and CG10200/hase und igel (hui) are expressed in A cells downstream of Hh signaling; and RNA interference for hui, Stubble, and CG16884 revealed that each is essential to wing development. We carried out a global screen for genes with compartment-specific expression using expression array hybridization to compare transcript levels in A and P wing disc cells. GFP-labeled wing imaginal discs (ptc-Gal4/UAS-GFP and hh-Gal4/UAS-GFP) were micro-dissected under a fluorescence dissecting microscope. RNA isolation, amplification and microarray procedures were previously described (Klebes et al., 2002; Klebes et al., 2005; Klebes and Kornberg, 2008). In brief, hybridization probes were generated by two rounds of T7-catalyzed linear RNA amplification and labeled with Cy3 and Cy5 dyes. Reciprocally labeled probes (M-bM-^@M-^Xdye flipM-bM-^@M-^Y) were hybridized to custom produced glass microarrays (GPL2581) that contained approximately 14,000 100-600 bp exon sequences that were generated by PCR. Signal intensities were collected with a GenePix 4000B Scanner and processed with GenePix software (Molecular Devices) and global median normalized with NOMAD (http://ucsf-nomad.sourceforge.net/).

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Ansgar Klebes 

PROVIDER: E-GEOD-46601 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress