Abstract
The Drosophila disconnected (disco) gene encodes a C2H2-type zinc finger transcription factor required for the development of the central and peripheral nervous systems. We report that disco participates in a positive feedback loop with the Dll gene, a master regulator of ventral appendage development. Dll function is not only required for proper disco expression in antenna and leg discs, but is also sufficient for ectopic expression of disco in the developing retina and wing imaginal discs. Conversely, disco gene function is required for the maintenance of Dll expression. We show that Dll phenotypes are partially rescued by the up-regulation of disco expression in the Dll domain. Reduction in disco gene function disrupts antenna and leg development, and the phenotypes closely resemble that produced by Dll alleles. These observations demonstrate that disco plays a fundamental role in the Dll-dependent patterning of antenna and leg, perhaps as a regulator of Dll gene expression.
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References
Abu-Shaar M, Mann RS (1998) Generation of multiple antagonistic domains along the proximodistal axis during Drosophila leg development. Development 126:3821–3830
Adachi-Yamada T, Harumoto T, Sakurai K, Ueda R, Saigo K, O’Connor MB, Nakato H (2005) Wing-to-leg homeosis by spineless causes apoptosis regulated by fish-lips, a novel leucine-rich repeat transmembrane protein. Mol Cell Biol 25:3140–3150
Ashburner M (1989) Drosophila: a laboratory handbook. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Basler K, Struhl G (1994) Compartment boundaries and the control of Drosophila limb pattern by Hedgehog protein. Nature 368:208–214
Bolinger RA, Boekhoff-Falk G (2005) Distal-less functions in subdividing the Drosophila thoracic limb primordium. Dev Dyn 232:801–816
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415
Brand AH, Manoukian AS, Perrimon N (1994) Ectopic expression in Drosophila. Methods Cell Biol 44:635–654
Calleja M, Moreno E, Pelaz S, Morata G (1996) Visualization of gene expression in living adult Drosophila. Science 274:252–255
Campbell G (2005) Regulation of gene expression in the distal region of the Drosophila leg by the Hox11 homolog, C15. Dev Biol 278:607–618
Campbell G, Tomlinson A (1995) Initiation of the proximo-distal axis in insect legs. Development 12:619–628
Campbell G, Tomlinson A (1998) The roles of the homeobox genes aristaless and distal-less in patterning the legs and wings of Drosophila. Development 125:4483–4493
Campbell G, Weaver T, Tomlinson A (1993) Axis specification in the developing Drosophila appendage: the role of wingless, decapentaplegic, and the homeobox gene aristaless. Cell 74:1113–1123
Campos AR, Fischbach KF, Steller H (1992) Survival of photoreceptor neurons in the compound eye of Drosophila depends on connections with the optic ganglia. Development 114:355–366
Campos AR, Lee KJ, Steller H (1995) Establishment of neuronal connectivity during development of the Drosophila larval visual system. J Neurobiol 283:13–29
Chen R, Amoui M, Zhang Z, Mardon G (1997) Dachshund and eyes absent proteins form a complex and function synergistically to induce ectopic eye development in Drosophila. Cell 91:893–903
Chu J, Dong PD, Panganiban G (2002) Limb type-specific regulation of bric a brac contributes to morphological diversity. Development 129:695–704
Cohen SM (1990) Specification of limb development in the Drosophila embryo by positional cues from segmentation genes. Nature 343:173–177
Cohen SM, Jurgen G (1989) Proximal-distal pattern formation in Drosophila: cell autonomous requirement for distal-less gene activity in limb development. EMBO J 8:2045–2055
Cohen SM, Bronner G, Kuttner F, Jurgens G, Jackle H (1989) Distal-less encodes a homoeodomain protein required for limb development in Drosophila. Nature 338:432–434
Cohen B, Wimmer EA, Cohen SM (1991) Early development of leg and wing primordia in the Drosophila embryo. Mech Dev 33:229–240
Collins RT, Treisman JE (2000) Osa-containing Brahma chromatin remodeling complexes are required for the repression of wingless target genes. Genes Dev 14:3140–3152
Diaz-Benjumea FJ, Cohen B, Cohen SM (1994) Cell interaction between compartments establishes the proximal-distal axis of Drosophila legs. Nature 372:175–179
Dominguez M, Casare F (2005) Organ specification-growth control connection: new in-sights from the Drosophila eye-antennal disc. Dev Dyn 232:673–684
Dong PDS, Chu J, Panganiban G (2000) Coexpression of the homeobox genes distal-less and homothorax determines Drosophila antennal identity. Development 127:209–216
Dong PD, Chu J, Panganiban G (2001) Proximodistal domain specification and interactions in developing Drosophila appendages. Development 128:2365–2372
Dong PDS, Dicks JS, Panganiban G (2002) Distal-less and homothorax regulate multiple targets to pattern the Drosophila antenna. Development 129:1967–1974
Duncan DM, Burgess EA, Duncan I (1998) Control of distal antennal identity and tarsal development in Drosophila by spineless-aristapedia, a homolog of the mammalian dioxin receptor. Genes Dev 12:1290–1303
Emerald BS, Curtiss J, Mlodzik M, Cohen SM (2003) Distal antenna and distal antenna related encode nuclear proteins containing pipsqueak motifs involved in antenna development in Drosophila. Development 130:1171–1180
Fischbach K, Heisenberg M (1984) Neurogenetics and behavior in insects. J Expt Biol 112:65–93
Glossop NR, Shepherd D (1998) disconnected mutants show disruption to the central projections of proprioceptive neurons in Drosophila melanogaster. J Neurobiol 36:337–347
Godt D, Couderc JL, Cramton SE, Laski FA (1993) Pattern formation in the limbs of Drosophila: bric á brac is expressed in both a gradient and a wave-like pattern and is required for specification and proper segmentation of the tarsus. Development 119:799–812
Gorfinkiel N, Morata G, Guerrero I (1997) The homeobox gene distal-less induces ventral appendage development in Drosophila. Genes Dev 11:2259–2271
Gorfinkiel N, Sanchez L, Guerrero I (1999) Drosophila terminalia as an appendage-like structure. Mech Dev 868:113–123
Goto S, Hayashi S (1997) Specification of the embryonic limb primordium by graded activity of decapentaplegic. Development 124:125–132
Hardin PE, Hall JC, Rosbash M (1992) Behavioral and molecular analyses suggest that circadian output is disrupted by disconnected mutants in D. melanogaster. EMBO J 11:1–6
Hay BA, Maile R, Rubin GM (1997) P element insertion-dependent gene activation in the Drosophila eye. Proc Natl Acad Sci USA 94:5195–5200
Heisenberg M, Bohl K (1979) Isolation of anatomical brain mutants of Drosophila by histochemical means. Z Naturforsch 34:143–147
Heilig JS, Freeman M, Laverty T, Lee KJ, Campos AR, Rubin GM, Steller H (1991) Isolation and characterization of the disconnected gene of Drosophila melanogaster. EMBO J 10:809–815
Helfrich-Forster C (1998) Robust circadian rhythmicity of Drosophila melanogaster requires the presence of lateral neurons: a brain-behavioral study of disconnected mutants. J Comp Physiol 182:435–453
Ibeas JM, Bray SJ (2003) Bowl is required downstream of Notch for elaboration of distal limb patterning. Development 130:5943–5952
Kojima T (2004) The mechanism of Drosophila leg development along the proximodistal axis. Dev Growth Differ 46:115–129
KojimaT SM, Saigo K (2000) Formation and specification of distal leg segments in Drosophila by dual Bar homeobox genes, BarH1 and BarH2. Development 127:769–778
Kojima T, Tsuji T, Saigo K (2005) A concerted action of a paired-type homeobox gene, aristaless, and a homolog of Hox11/tlx homeobox gene, clawless, is essential for the distal tip development of the Drosophila leg. Dev Biol 279:434–445
Lee KJ, Freeman M, Steller H (1991) Expression of the disconnected gene during development of Drosophila melanogaster. EMBO J 10:817–826
Lee KJ, Mukhopadhyay M, Pelka P, Campos AR, Steller H (1999) Autoregulation of the Drosophila disconnected gene in the developing visual system. Dev Biol 214:385–398
Lee YS, Carthew RW (2003) Making a better RNAi vector for Drosophila: use of intron spacers. Methods 30:322–329
Mahaffey JW, Griswold CM, Cao QM (2001) The Drosophila genes disconnected and disco-related are redundant with respect to larval head development and accumulation of mRNAs from deformed target genes. Genetics 15:225–236
Mukhopadhyay M, Campos AR (1995) The larval optic nerve is required for the development of an identified serotonergic arborization in Drosophila melanogaster. Dev Biol 169:629–643
Panganiban G, Sebring A, Nagy L, Carroll S (1995) The development of crustacean limbs and the evolution of arthropods. Science 270:1363–1366
Patel M, Farzana L, Robertson LK, Hutchinson J, Grubbs N, Shepherd MN, Mahaffey JW (2007) The appendage role of insect disco genes and possible implications on the evolution of the maggot larval form. Dev Biol 309:56–69
Quiring R, Walldorf U, Kloter U, Gehring WJ (1994) Homology of the eyeless gene of Drosophila to the small eye gene in mice and aniridia in humans. Science 265:785–789
Raz E, Shilo B (1993) Establishment of ventral cell fates in the Drosophila embryonic ectoderm requires DER, the EGF receptor homolog. Genes Dev 7:1937–1948
Robertson LK, Bowling DB, Mahaffey JP, Imiolczyk B, Mahaffey JW (2004) An interactive network of zinc-finger proteins contributes to regionalization of the Drosophila embryo and establishes the domains of HOM-C protein function. Development 131:2781–2789
Sanders LR, Patel M, Mahaffey JW (2008) The Drosophila gap gene giant has an anterior segment identity function mediated through disconnected and teashirt. Genetics 179:441–453
Sato T (1984) A new homeotic mutation affecting antenna and legs. Drosophila Information Service 60:180–182
Schock F, Joachim Reischl J, Wimmer E, Taubert H, Beverly A, Purnell BA, Jackle H (2000) Phenotypic suppression of empty spiracles is prevented by buttonhead. Nature 405:351–354
Spradling AC, Rubin GM (1982) Transposition of cloned P elements into Drosophila germ line chromosomes. Science 218:341–347
Steller H, Fischbach KF, Rubin GM (1987) disconnected: a locus required for neuronal pathway formation in the visual system of Drosophila. Cell 50:1139–1153
Sunkel CE, Whittle JRS (1987) Brista: a gene involved in the specification and differentiation of distal cephalic and thoracic structures in Drosophila melanogaster. Wilhelm Roux Arch Dev Biol 196:124–132
Wu J, Cohen SM (1999) Proximodistal axis formation in the Drosophila leg: subdivision into proximal and distal domains by homothorax and distal-less. Development 126:109–117
Acknowledgements
We thank Drs. Gerard Campbell, Grace Boekhoff-Falk, Ian Duncan, Isabel Guerrero, Frank Laski, University of Iowa Developmental Studies Hybridoma Bank, and Bloomington Drosophila Stock Center for kindly providing the antibodies and fly lines used in this study. We are also grateful to Dr. Richard Carthew for the pWIZ RNAi vector and to Drs. Andre Bedard, Roger Jacobs and Anindya Dutta for critical reading of the manuscript.
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This work is supported by a grant from the Natural Sciences and Engineering Research Council of Canada (NSERC) to A.R. Campos.
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Suppl. Fig. 1
a Endogenous disco mRNA is gradually down-regulated when UAS-disco RNAi is driven by HSP70-GAL4. Embryos of genotype HSP70-GAL4; UAS-disco RNAi, aged 0–14 h, were heat-shocked for 1 h at 37°C and allowed to recover at 25°C for 1, 2, or 3 h. Whole-mount in situ hybridization was carried out (see: Tautz D, Pfeifle C [1989] A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback.Chromosoma 98:81–85) with hybridization probes generated from a discocDNA cloned into Bluescript vector by using the DIG RNA Labeling Kit of Roche AppliedSciences (Top a–c stage-matched non-heat-shocked controls, Bottom d–f heat-shocked embryos). b Endogenous disco level of heat-shocked HSP70-GAL4; UAS-disco RNAi embryos was measured by real-time PCR. Embryos carrying HSP70-GAL4; UAS-disco RNAi elements and stage-matched OR control embryos, aged 0–14 h, were heat-shocked for 1 h at 37°C and allowed to recover at 25°C for 1, 2, or 3 h. RNA was isolated and real-time PCR was carried out with DNA Master SYBR green-1. An approximately two-, four-, and nine-fold reduction of disco RNA level was observed 1, 2, and 3 h, respectively, after heat-shock-induced expression of UAS-disco RNAi transgenes compared with that of wild type (OR) embryos. (DOC 1000 kb)
Suppl. Fig. 2
Expression of UAS-disco RNAi suppresses the phenotype of GMR-GAL4; UAS-disco. Environmental scanning electron micrographs of Drosophila compound eyes (a–d are shown at higher magnification in e–h). a, e Wild type. b, f GMR-GAL4; UAS-disco. Ectopic expression of UAS-disco posterior to the morphogenetic furrow by using the GMR-GAL4 driver causes small rough eyes lacking eye pigments. The lenses of the cone cells appear fused, the interommatidial sensory bristles are displaced and/or missing, and many tiny bristles of unknown origin are present. c, g GMR-GAL4; UAS-disco/UAS-disco RNAi. This phenotype is completely rescued by co-expressing UAS-disco RNAi. d, h GMR-GAL4; UAS-disco RNAi shows a normal eye. (DOC 3330 kb)
Suppl. Fig. 3
Function of disco is not required for proper expression of DAC in leg discs. a Wild type. b Dll-GAL4/CyO. c disco 1/disco 1. d Dll-GAL4; UAS-disco RNAi. (DOC 433 kb)
Suppl. Table 1
Dll alleles increase the lethality of disco 1. Dll 5, Dll9, and Dll 3 in combination with disco 1 hemizygous flies exhibit 100%, 75%, and 52.38% lethality, respectively, compared with disco 1 hemizygous alone, whereas the hth 0475 allele has no impact on lethality. (DOC 33 kb)
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Dey, B.K., Zhao, XL., Popo-Ola, E. et al. Mutual regulation of the Drosophila disconnected (disco) and Distal-less (Dll) genes contributes to proximal-distal patterning of antenna and leg. Cell Tissue Res 338, 227–240 (2009). https://doi.org/10.1007/s00441-009-0865-z
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DOI: https://doi.org/10.1007/s00441-009-0865-z