A Systematic Screen for Tube Morphogenesis and Branching Genes in the Tracheal System


Many signaling proteins and transcription factors that induce and pattern organs have been identified, but relatively few of the downstream effectors that execute morphogenesis programs. Because such morphogenesis genes may function in many organs and developmental processes, mutations in them are expected to be pleiotropic and hence ignored or discarded in most standard genetic screens. Here we describe a systematic screen designed to identify all Drosophila third chromosome genes (∼40% of the genome) that function in development of the tracheal system, a tubular respiratory organ that provides a paradigm for branching morphogenesis. To identify potentially pleiotropic morphogenesis genes, the screen included analysis of marked clones of homozygous mutant tracheal cells in heterozygous animals, plus a secondary screen to exclude mutations in general “house-keeping” genes. From a collection including more than 5,000 lethal mutations, we identified 133 mutations representing ∼70 or more genes that subdivide the tracheal terminal branching program into six genetically separable steps, a previously established cell specification step plus five major morphogenesis and maturation steps: branching, growth, tubulogenesis, gas-filling, and maintenance. Molecular identification of 14 of the 70 genes demonstrates that they include six previously known tracheal genes, each with a novel function revealed by clonal analysis, and two well-known growth suppressors that establish an integral role for cell growth control in branching morphogenesis. The rest are new tracheal genes that function in morphogenesis and maturation, many through cytoskeletal and secretory pathways. The results suggest systematic genetic screens that include clonal analysis can elucidate the full organogenesis program and that over 200 patterning and morphogenesis genes are required to build even a relatively simple organ such as the Drosophila tracheal system.


Vyšlo v časopise: A Systematic Screen for Tube Morphogenesis and Branching Genes in the Tracheal System. PLoS Genet 7(7): e32767. doi:10.1371/journal.pgen.1002087
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002087

Souhrn

Many signaling proteins and transcription factors that induce and pattern organs have been identified, but relatively few of the downstream effectors that execute morphogenesis programs. Because such morphogenesis genes may function in many organs and developmental processes, mutations in them are expected to be pleiotropic and hence ignored or discarded in most standard genetic screens. Here we describe a systematic screen designed to identify all Drosophila third chromosome genes (∼40% of the genome) that function in development of the tracheal system, a tubular respiratory organ that provides a paradigm for branching morphogenesis. To identify potentially pleiotropic morphogenesis genes, the screen included analysis of marked clones of homozygous mutant tracheal cells in heterozygous animals, plus a secondary screen to exclude mutations in general “house-keeping” genes. From a collection including more than 5,000 lethal mutations, we identified 133 mutations representing ∼70 or more genes that subdivide the tracheal terminal branching program into six genetically separable steps, a previously established cell specification step plus five major morphogenesis and maturation steps: branching, growth, tubulogenesis, gas-filling, and maintenance. Molecular identification of 14 of the 70 genes demonstrates that they include six previously known tracheal genes, each with a novel function revealed by clonal analysis, and two well-known growth suppressors that establish an integral role for cell growth control in branching morphogenesis. The rest are new tracheal genes that function in morphogenesis and maturation, many through cytoskeletal and secretory pathways. The results suggest systematic genetic screens that include clonal analysis can elucidate the full organogenesis program and that over 200 patterning and morphogenesis genes are required to build even a relatively simple organ such as the Drosophila tracheal system.


Zdroje

1. AffolterMBellusciSItohNShiloBThieryJP 2003 Tube or not tube: remodeling epithelial tissues by branching morphogenesis. Dev Cell 4 11 18

2. HoganBLKolodziejPA 2002 Organogenesis: molecular mechanisms of tubulogenesis. Nat Rev Genet 3 513 523

3. LubarskyBKrasnowMA 2003 Tube morphogenesis: making and shaping biological tubes. Cell 112 19 28

4. NelsonWJ 2003 Tube morphogenesis: closure, but many openings remain. Trends Cell Biol 13 615 621

5. WieschausEF 1996 From molecular patterns to morphogenesis-The lessons from studies on the fruit fly Drosophila (Nobel Lecture). Angewandte Chemie International Edition in English 35 2188 2194

6. SpradlingACSternDBeatonARhemEJLavertyT 1999 The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes. Genetics 153 135 177

7. MetzsteinMMKrasnowMA 2006 Functions of the nonsense-mediated mRNA decay pathway in Drosophila development. PLoS Genet 2 e180

8. ManningGKrasnowMA 1993 Development of the Drosophila tracheal system. BateMMartinez AriasA The Development of Drosophila melanogaster Cold Spring Harbor, NY Cold Spring Harbor Press 609 685

9. GhabrialALuschnigSMetzsteinMMKrasnowMA 2003 Branching morphogenesis of the Drosophila tracheal system. Annu Rev Cell Dev Biol 19 623 647

10. CabernardCNeumannMAffolterM 2004 Cellular and molecular mechanisms involved in branching morphogenesis of the Drosophila tracheal system. J Appl Physiol 97 2347 2353

11. SamakovlisCHacohenNManningGSutherlandDCGuilleminK 1996 Development of the Drosophila tracheal system occurs by a series of morphologically distinct but genetically coupled branching events. Development 122 1395 1407

12. SamakovlisCManningGStenebergPHacohenNCanteraR 1996 Genetic control of epithelial tube fusion during Drosophila tracheal development. Development 122 3531 3536

13. GhabrialASKrasnowMA 2006 Social interactions among epithelial cells during tracheal branching morphogenesis. Nature 441 746 749

14. GuilleminKGroppeJDuckerKTreismanRHafenE 1996 The pruned gene encodes the Drosophila serum response factor and regulates cytoplasmic outgrowth during terminal branching of the tracheal system. Development 122 1353 1362

15. RibeiroCNeumannMAffolterM 2004 Genetic control of cell intercalation during tracheal morphogenesis in Drosophila. Curr Biol 14 2197 2207

16. TsarouhasVSentiKAJayaramSATiklovaKHemphalaJ 2007 Sequential pulses of apical epithelial secretion and endocytosis drive airway maturation in Drosophila. Dev Cell 13 214 225

17. WigglesworthVB 1983 The physiology of insect tracheoles. Adv Insect Physiol 17 88 148

18. BarTGuldnerFHWolffJR 1984 “Seamless” endothelial cells of blood capillaries. Cell Tissue Res 235 99 106

19. KameiMSaundersWBBaylessKJDyeLDavisGE 2006 Endothelial tubes assemble from intracellular vacuoles in vivo. Nature 442 453 456

20. YoshidaYYamadaMWakabayashiKIkutaFKumanishiT 1989 Endothelial basement membrane and seamless-type endothelium in the repair process of cerebral infarction in rats. Virchows Arch A Pathol Anat Histopathol 414 385 392

21. BerryKLBulowHEHallDHHobertO 2003 A C. elegans CLIC-like protein required for intracellular tube formation and maintenance. Science 302 2134 2137

22. JurgensGKludingGNusslein-VolhardCWieschausE 1984 Mutations affecting the pattern of the larval cuticle in Drosophila melanogaster. II. Zygotic loci on the third chromosome. Roux's Archives of Developmental Biology 193 283 295

23. IsaacDDAndrewDJ 1996 Tubulogenesis in Drosophila: a requirement for the trachealess gene product. Genes Dev 10 103 117

24. WilkRWeizmanIShiloBZ 1996 trachealess encodes a bHLH-PAS protein that is an inducer of tracheal cell fates in Drosophila. Genes Dev 10 93 102

25. GlazerLShiloBZ 1991 The Drosophila FGF-R homolog is expressed in the embryonic tracheal system and appears to be required for directed tracheal cell extension. Genes Dev 5 697 705

26. KlambtCGlazerLShiloBZ 1992 breathless, a Drosophila FGF receptor homolog, is essential for migration of tracheal and specific midline glial cells. Genes Dev 6 1668 1678

27. ShishidoEHigashijimaSEmoriYSaigoK 1993 Two FGF-receptor homologues of Drosophila: one is expressed in mesodermal primordium in early embryos. Development 117 751 761

28. BeitelGJKrasnowMA 2000 Genetic control of epithelial tube size in the Drosophila tracheal system. Development 127 3271 3282

29. SutherlandDSamakovlisCKrasnowMA 1996 branchless encodes a Drosophila FGF homolog that controls tracheal cell migration and the pattern of branching. Cell 87 1091 1101

30. IkeyaTHayashiS 1999 Interplay of Notch and FGF signaling restricts cell fate and MAPK activation in the Drosophila trachea. Development 126 4455 4463

31. LinXBuffEMPerrimonNMichelsonAM 1999 Heparan sulfate proteoglycans are essential for FGF receptor signaling during Drosophila embryonic development. Development 126 3715 3723

32. ChiharaTHayashiS 2000 Control of tracheal tubulogenesis by Wingless signaling. Development 127 4433 4442

33. LlimargasM 2000 Wingless and its signalling pathway have common and separable functions during tracheal development. Development 127 4407 4417

34. MyatMMLightfootHWangPAndrewDJ 2005 A molecular link between FGF and Dpp signaling in branch-specific migration of the Drosophila trachea. Dev Biol 281 38 52

35. BaerMMBilsteinALeptinM 2007 A clonal genetic screen for mutants causing defects in larval tracheal morphogenesis in Drosophila. Genetics 176 2279 2291

36. Chanut-DelalandeHJungACLinLBaerMMBilsteinA 2007 A genetic mosaic analysis with a repressible cell marker screen to identify genes involved in tracheal cell migration during Drosophila air sac morphogenesis. Genetics 176 2177 2187

37. JungACRibeiroCMichautLCertaUAffolterM 2006 Polychaetoid/ZO-1 is required for cell specification and rearrangement during Drosophila tracheal morphogenesis. Curr Biol 16 1224 1231

38. LuschnigSBatzTArmbrusterKKrasnowMA 2006 serpentine and vermiform encode matrix proteins with chitin binding and deacetylation domains that limit tracheal tube length in Drosophila. Curr Biol 16 186 194

39. StahlMSchuhRAdryanB 2007 Identification of FGF-dependent genes in the Drosophila tracheal system. Gene Expr Patterns 7 202 209

40. ZhuMYWilsonRLeptinM 2005 A screen for genes that influence fibroblast growth factor signal transduction in Drosophila. Genetics 170 767 777

41. AdamsMDCelnikerSEHoltRAEvansCAGocayneJD 2000 The genome sequence of Drosophila melanogaster. Science 287 2185 2195

42. LeeTLuoL 2001 Mosaic analysis with a repressible cell marker (MARCM) for Drosophila neural development. Trends Neurosci 24 251 254

43. StowersRSSchwarzTL 1999 A genetic method for generating Drosophila eyes composed exclusively of mitotic clones of a single genotype. Genetics 152 1631 1639

44. AffolterMNellenDNussbaumerUBaslerK 1994 Multiple requirements for the receptor serine/threonine kinase thick veins reveal novel functions of TGF beta homologs during Drosophila embryogenesis. Development 120 3105 3117

45. JareckiJJohnsonEKrasnowMA 1999 Oxygen regulation of airway branching in Drosophila is mediated by branchless FGF. Cell 99 211 220

46. LeviBPGhabrialASKrasnowMA 2006 Drosophila talin and integrin genes are required for maintenance of tracheal terminal branches and luminal organization. Development 133 2383 2393

47. SwansonLEBeitelGJ 2006 Tubulogenesis: an inside job. Curr Biol 16 R51 53

48. JeonMZinnK 2009 Receptor tyrosine phosphatases control tracheal tube geometries through negative regulation of Egfr signaling. Development 136 3121 3129

49. ForsterDArmbrusterKLuschnigS 2009 Sec24-dependent secretion drives cell-autonomous expansion of tracheal tubes in Drosophila. Curr Biol 20 62 68

50. BlumRStephensDJSchulzI 2000 Lumenal targeted GFP, used as a marker of soluble cargo, visualises rapid ERGIC to Golgi traffic by a tubulo-vesicular network. J Cell Sci 113 3151-3159

51. GrueberWBYeBMooreAWJanLYJanYN 2003 Dendrites of distinct classes of Drosophila sensory neurons show different capacities for homotypic repulsion. Curr Biol 13 618 626

52. MobergKHSchelbleSBurdickSKHariharanIK 2005 Mutations in erupted, the Drosophila ortholog of mammalian tumor susceptibility gene 101, elicit non-cell-autonomous overgrowth. Dev Cell 9 699 710

53. DevineWPLubarskyBShawKLuschnigSMessinaL 2005 Requirement for chitin biosynthesis in epithelial tube morphogenesis. Proc Natl Acad Sci U S A 102 17014 17019

54. MoussianBTangETonningAHelmsSSchwarzH 2006 Drosophila Knickkopf and Retroactive are needed for epithelial tube growth and cuticle differentiation through their specific requirement for chitin filament organization. Development 133 163 171

55. TaponNItoNDicksonBJTreismanJEHariharanIK 2001 The Drosophila tuberous sclerosis complex gene homologs restrict cell growth and cell proliferation. Cell 105 345 355

56. CentaninLDekantyARomeroNIrisarriMGorrTA 2008 Cell autonomy of HIF effects in Drosophila: tracheal cells sense hypoxia and induce terminal branch sprouting. Dev Cell 14 547 558

57. JusticeRWZilianOWoodsDFNollMBryantPJ 1995 The Drosophila tumor suppressor gene warts encodes a homolog of human myotonic dystrophy kinase and is required for the control of cell shape and proliferation. Genes Dev 9 534 546

58. XuTWangWZhangSStewartRAYuW 1995 Identifying tumor suppressors in genetic mosaics: the Drosophila lats gene encodes a putative protein kinase. Development 121 1053 1063

59. KlambtC 1993 The Drosophila gene pointed encodes two ETS-like proteins which are involved in the development of the midline glial cells. Development 117 163 176

60. AraujoSJAslamHTearGCasanovaJ 2005 mummy/cystic encodes an enzyme required for chitin and glycan synthesis, involved in trachea, embryonic cuticle and CNS development--analysis of its role in Drosophila tracheal morphogenesis. Dev Biol 288 179 193

61. TonningAHemphalaJTangENannmarkUSamakovlisC 2005 A transient luminal chitinous matrix is required to model epithelial tube diameter in the Drosophila trachea. Dev Cell 9 423 430

62. BrownNHGregorySLRickollWLFesslerLIProutM 2002 Talin is essential for integrin function in Drosophila. Dev Cell 3 569 579

63. WuLNiemeyerBColleyNSocolichMZukerCS 1995 Regulation of PLC-mediated signalling in vivo by CDP-diacylglycerol synthase. Nature 373 216 222

64. MortimerNTMobergKH 2009 Regulation of Drosophila embryonic tracheogenesis by dVHL and hypoxia. Dev Biol 329 294 305

65. PotterCJHuangHXuT 2001 Drosophila Tsc1 functions with Tsc2 to antagonize insulin signaling in regulating cell growth, cell proliferation, and organ size. Cell 105 357 368

66. LoweSLPeterFSubramaniamVNWongSHHongW 1997 A SNARE involved in protein transport through the Golgi apparatus. Nature 389 881 884

67. HayJCChaoDSKuoCSSchellerRH 1997 Protein interactions regulating vesicle transport between the endoplasmic reticulum and Golgi apparatus in mammalian cells. Cell 89 149 158

68. GipsSJKandzariDEGoldschmidt-ClermontPJ 1994 Growth factor receptors, phospholipases, phospholipid kinases and actin reorganization. Semin Cell Biol 5 201 208

69. JayaramSASentiKATiklovaKTsarouhasVHemphalaJ 2008 COPI vesicle transport is a common requirement for tube expansion in Drosophila. PLoS One 3 e1964

70. SternlichtHFarrGWSternlichtMLDriscollJKWillisonK 1993 The t-complex polypeptide 1 complex is a chaperonin for tubulin and actin in vivo. Proc Natl Acad Sci U S A 90 9422 9426

71. NechiporukTFernandezTEVasioukhinV 2007 Failure of epithelial tube maintenance causes hydrocephalus and renal cysts in Dlg5-/- mice. Dev Cell 13 338 350

72. GenesteOCopelandJWTreismanR 2002 LIM kinase and Diaphanous cooperate to regulate serum response factor and actin dynamics. J Cell Biol 157 831 838

73. HauckeV 2005 Phosphoinositide regulation of clathrin-mediated endocytosis. Biochem Soc Trans 33 1285 1289

74. Martin-BelmonteFGassamaADattaAYuWRescherU 2007 PTEN-mediated apical segregation of phosphoinositides controls epithelial morphogenesis through Cdc42. Cell 128 383 397

75. MangoSE 2009 The molecular basis of organ formation: insights from the C. elegans foregut. Annu Rev Cell Dev Biol 25 597 628

76. HemphalaJUvACanteraRBraySSamakovlisC 2003 Grainy head controls apical membrane growth and tube elongation in response to Branchless/FGF signalling. Development 130 249 258

77. GaudetJMuttumuSHornerMMangoSE 2004 Whole-genome analysis of temporal gene expression during foregut development. PLoS Biol 2 e352

78. JonssonJCarlssonLEdlundTEdlundH 1994 Insulin-promoter-factor 1 is required for pancreas development in mice. Nature 371 606 609

79. SvenssonPWilliamsCLundebergJRydenPBergqvistI 2007 Gene array identification of Ipf1/Pdx1-/- regulated genes in pancreatic progenitor cells. BMC Dev Biol 7 129

80. BrandAHPerrimonN 1993 Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118 401 415

81. ShigaYTanaka-MatakatsuMHayashiS 1996 A nuclear GFP/β-galactosidase fusion protein as a marker for morphogenesis in living Drosophila. Dev Growth Differ 38 99 106

82. AshburnerM 1989 Drosophila: A laboratory handbook. Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press

83. BergerJSuzukiTSentiKAStubbsJSchaffnerG 2001 Genetic mapping with SNP markers in Drosophila. Nat Genet 29 475 481

84. MartinSGDobiKCSt JohnstonD 2001 A rapid method to map mutations in Drosophila. Genome Biol 2 RESEARCH0036

85. LeeTHacohenNKrasnowMMontellDJ 1996 Regulated Breathless receptor tyrosine kinase activity required to pattern cell migration and branching in the Drosophila tracheal system. Genes Dev 10 2912 2921

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