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doi: 10.1242/10.1242/dev.00293

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Discovery of genes with highly restricted expression patterns in the Drosophila wing disc using DNA oligonucleotide microarrays

Miranda J. Butler1, Thomas L. Jacobsen2, Donna M. Cain2, Michael G. Jarman1, Michael Hubank3, J. Robert S. Whittle1, Roger Phillips1,*,{dagger} and Amanda Simcox1,2,*,{dagger}

1 School of Biological Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK
2 Department of Molecular Genetics, 484 West 12th Avenue, The Ohio State University, Columbus, OH 43210, USA
3 Institute of Child Health, University College, London, 30 Guilford Street, London WC1N 1EH, UK



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  		Fig. 1. Drosophila wing disc and fate maps. (A) Third instar wing disc. To generate the wing/hinge and body wall fragments, the discs were cut across above the folds corresponding to the hinge region (arrowheads). (B) Fate map of the wing disc showing the anterior-posterior (AP) and dorsal-ventral (DV) compartment boundaries and major regions in the disc. In the adult, the wing pouch (green) gives rise to the wing blade, the hinge (yellow) constricts to form a mobile link to the body wall (blue) or mesonotum of the fly. (C) Cell layers of the wing disc. There are three cell layers: the squamous epithelium or peripodial membrane, the columnar epithelium that gives rise to the adult epidermis, and the adepithial layer comprised of myoblasts, which develop into the flight muscles of the thorax, and tracheal cells of the larval and future adult airways.

 


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  		Fig. 2. Expression patterns of transcripts enriched in the body wall sample. The expression patterns of the transcripts are shown in the overall rank order according to fold change with the most highly enriched shown first (Table 1). (A) tailup (tup) is expressed in cells of the dorsal posterior epithelium. (B) CG11100 is expressed in the myoblasts. (C) CG15064 is expressed in the myoblasts. (D) CG15353 is expressed in tracheal cells. (E) CG6921 is highly expressed at the proximal tip of the disc and weakly in tracheal cells. (F) BM-40/SPARC is expressed in the myoblasts. (G) Obp56a/CG11797 is expressed in stalk cells at the proximal tip of the disc. (H) zinc finger homology 1 (zfh1) is expressed in the myoblasts. (I) viking (vkg) is expressed in myoblasts and blood cells (arrowheads, one group of cells is out of the plane of focus). (J) Elongation factor 1 alpha100E (Ef1 alpha) is expressed in the myoblasts. (K) Obp99a/CG18111 is expressed in the epithelium in cells of the presumptive scutellum. (L) CG10126 is expressed in stalk cells at the proximal tip of the disc. (M) CG9338 is expressed in tracheal cells. (N) Cg25C is expressed in myoblasts and blood cells (arrowhead). (O) Actin 57B (Act57B) is expressed in myoblasts. (P) CG5397 is expressed in tracheal cells and blood cells (arrowhead). (Q) Imaginal disc growth factor 4 (Idgf4) is expressed in the dorsal peripodial membrane. (R) CG4386 is expressed in the dorsal branch of the tracheal system. (S) CG3244 is expressed in two patches of cells in the prescutum (the smaller cluster is indicated with an arrowhead). (T) CG2663 is expressed in the trachea. (U) CG10275 is expressed in the myoblasts, hinge columnar epithelium (white arrowhead), and peripodial membrane (black arrowhead). (V) CG8689 is expressed in the myoblasts. (W) thrombospondin/CG11326 (tsp) is expressed in the dorsal posterior body-wall region and the anterior ventral wing hinge. (X) Glutamine synthetase (GS2) is expressed in tracheal cells. (Y) Glucose dehydrogenase (Gld) is expressed in the posterior dorsal epithelium in cells of the presumptive postnotum and a line of cells in the hinge region (arrowhead).

 


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  		Fig. 3. Expression patterns of transcripts enriched in the wing/hinge sample. The expression patterns of the transcripts are shown in the overall rank order according to fold change with the most highly enriched shown first (Table 2). All genes are expressed in the columnar epithelium except where noted. (A) CG17278 is strongly expressed throughout the wing pouch, but not in cells at the DV margin, and expressed in a small cluster of cells in the dorsal hinge. (B) defective proventriculous (dve) is weakly expressed throughout the wing pouch, but not in cells at the DV margin. (C) POU domain protein 2 (pdm2) is expressed throughout the wing pouch. Expression levels vary with higher expression in cells probably corresponding to vein precursors. (D) CG8780 is expressed in cells of the mesopleura and ventral hinge. (E) ß-galactosidase/CG3132 (ß-gal) is expressed in a cluster of cells in the dorsal hinge and in blood cells that adhere to the anterior ventral margin of the disc. (F) Nep1/CG5894 is expressed in the mesopleura (arrowhead) and dorsal and ventral hinge. (G) odd paired (opa) is expressed in the mesopleura (arrowhead) and dorsal and ventral hinge. (H) CG15001 is weakly expressed throughout the wing pouch. (I) CG15489 is expressed weakly throughout the wing pouch. Expression levels vary with higher expression in cells probably corresponding to vein precursors. (J) Cyp310a1 (Cyp) is expressed throughout the wing pouch except in cells at the DV and AP margins. (K) CG15488 is expressed throughout the wing pouch. Expression levels vary with higher expression in cells probably corresponding to vein precursors. (L) CG15000 is weakly expressed throughout the wing pouch. (M) CG14534 is specifically expressed in cells that give rise to the posterior margin of the wing. (N) BG:DS00797.2/CG9008 (797.2) is strongly expressed throughout the wing pouch with some modulation in levels that does not appear to correlate with known features such as veins. Transcripts are also present at low levels in adeptihial cells (arrowhead, out of the plane of focus). (O) CG4861 is expressed very weakly throughout the wing pouch. (P) CG6469 is expressed in peripodial cells with higher levels of expression in ventral cells. (Q) CG14301 expression is weak but elevated expression was seen in cells at the anterior disc margin and in four clusters of cells in the wing pouch and overlying peripodial membrane. (R) Doc2/CG5187 is expressed in four patches of cells including a group of cells in the body wall (arrowhead). (S) UDP-glucosyl transferase 86Di/CG6658 (Ugt) is expressed in a diffuse pattern with elevated expression along the anterior disc margin, extending into the body wall region, and in intervein regions. (T) anachronism (ana) is expressed in clusters of cells and individual cells, probably neuroblasts. A cluster of cells in the body wall region express ana (arrowhead). (U) CG5758 is expressed in the hinge region. Expression is modulated so that the pattern has a patch-like appearance. (V) CG8381 is expressed strongly in the wing pouch with modulation at the DV margin and the presumptive veins. The gene is also expressed at low levels in several clusters of cells in the body wall region. (W) CG8483 is expressed along the DV margin and in several clusters of cells in the hinge and body wall regions. (X) Dorsocross 1 (Doc1) is expressed in four patches of cells including a group of cells in the body wall (arrowhead). (Y) wengen/CG6531 is expressed in the wing pouch with stronger expression in future veins and weakly in the body wall region (cells out of plane of focus).

 


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  		Fig. 4. Regulation by wg. In wild-type discs, CG17278 (A), Cyp310a1 (D) and CG8381 (G) are expressed broadly in the wing pouch but excluded from cells at the DV boundary. Misexpression of wg in the dorsal and ventral wing pouch (71B-GAL4; UAS-wg) inhibits expression of the genes in a broad band of cells in the wing pouch (B,E,H). Inhibition of Wg signaling at the DV margin (C96-GAL4; UAS-DNdTcf) induces more cells at the margin to express the genes (C,F,I). At the center of a wild-type disc approximately four rows of cells do not express CG17278 (A, arrowheads), whereas expressing cells are adjacent at this point when Wg is inhibited (C, arrowheads). When Wg is inhibited, Cyp310a1, expression spans the DV boundary for its entire length (F). Expression of CG8381 in the wild-type wing pouch is excluded from a band of six to seven cells at the DV boundary (G, arrowheads) and this is reduced to approximately two cells at the equivalent position when Wg is inhibited (I, arrowheads).

 




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