Ectopic Expression of Induces Spinal Defects, Urogenital Defects, and Anorectal Malformations in Mice
Danforth's short tail (Sd) is a semidominant mutation on mouse chromosome 2, characterized by spinal defects, urogenital defects, and anorectal malformations. However, the gene responsible for the Sd phenotype was unknown. In this study, we identified the molecular basis of the Sd mutation. By positional cloning, we identified the insertion of an early transposon in the Sd candidate locus approximately 12-kb upstream of Ptf1a. We found that insertion of the transposon caused overexpression of three neighboring genes, Gm13344, Gm13336, and Ptf1a, in Sd mutant embryos and that the Sd phenotype was not caused by disruption of an as-yet-unknown gene in the candidate locus. Using multiple knockout and knock-in mouse models, we demonstrated that misexpression of Ptf1a, but not of Gm13344 or Gm13336, in the notochord, hindgut, cloaca, and mesonephros was sufficient to replicate the Sd phenotype. The ectopic expression of Ptf1a in the caudal embryo resulted in attenuated expression of Cdx2 and its downstream target genes T, Wnt3a, and Cyp26a1; we conclude that this is the molecular basis of the Sd phenotype. Analysis of Sd mutant mice will provide insight into the development of the spinal column, anus, and kidney.
Vyšlo v časopise:
Ectopic Expression of Induces Spinal Defects, Urogenital Defects, and Anorectal Malformations in Mice. PLoS Genet 9(2): e32767. doi:10.1371/journal.pgen.1003204
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1003204
Souhrn
Danforth's short tail (Sd) is a semidominant mutation on mouse chromosome 2, characterized by spinal defects, urogenital defects, and anorectal malformations. However, the gene responsible for the Sd phenotype was unknown. In this study, we identified the molecular basis of the Sd mutation. By positional cloning, we identified the insertion of an early transposon in the Sd candidate locus approximately 12-kb upstream of Ptf1a. We found that insertion of the transposon caused overexpression of three neighboring genes, Gm13344, Gm13336, and Ptf1a, in Sd mutant embryos and that the Sd phenotype was not caused by disruption of an as-yet-unknown gene in the candidate locus. Using multiple knockout and knock-in mouse models, we demonstrated that misexpression of Ptf1a, but not of Gm13344 or Gm13336, in the notochord, hindgut, cloaca, and mesonephros was sufficient to replicate the Sd phenotype. The ectopic expression of Ptf1a in the caudal embryo resulted in attenuated expression of Cdx2 and its downstream target genes T, Wnt3a, and Cyp26a1; we conclude that this is the molecular basis of the Sd phenotype. Analysis of Sd mutant mice will provide insight into the development of the spinal column, anus, and kidney.
Zdroje
1. DunnLC (1940) A new mutation in the mouse affecting spinal column and urogenital system. Journal of heredity 31: 343–348.
2. GrünebergH (1958) Genetical studies on the skeleton of the mouse. XXII. The development of Danforth's short-tail. J Embryol Exp Morphol 6: 124–148.
3. FavreA, BrianoS, MazzolaC, BrizzolaraA, TorreM, et al. (1999) Anorectal malformations associated with enteric dysganglionosis in Danforth's short tail (Sd) mice. J Pediatr Surg 34: 1818–1821.
4. Gluecksohn-SchoenheimerS (1943) The morphological manifestations of a dominant mutation in mice affecting tail and urogenital system. Genetics 28: 341–348.
5. Gluecksohn-SchoenheimerS (1945) The Embryonic development of mutants of the Sd-strain in mice. Genetics 30: 29–38.
6. GrünebergH (1953) Genetical studies on the skeleton of the mouse. VI. Danforth's short-tail. J Genet 51: 317–326.
7. AndoT, SembaK, SudaH, SeiA, MizutaH, et al. (2011) The floor plate is sufficient for development of the sclerotome and spine without the notochord. Mech Dev 128: 129–140.
8. KosekiH, WallinJ, WiltingJ, MizutaniY, KispertA, et al. (1993) A role for Pax-1 as a mediator of notochordal signals during the dorsoventral specification of vertebrae. Development 119: 649–660.
9. Brand-SaberiB, EbenspergerC, WiltingJ, BallingR, ChristB (1993) The ventralizing effect of the notochord on somite differentiation in chick embryos. Anat Embryol (Berl) 188: 239–245.
10. EbenspergerC, WiltingJ, Brand-SaberiB, MizutaniY, ChristB, et al. (1995) Pax-1, a regulator of sclerotome development is induced by notochord and floor plate signals in avian embryos. Anat Embryol (Berl) 191: 297–310.
11. ZachgoJ, KornR, GosslerA (1998) Genetic interactions suggest that Danforth's short tail (Sd) is a gain-of-function mutation. Dev Genet 23: 86–96.
12. LanePW (1993) BirkenmeierCS (1993) Urogenital syndrome (us): a developmental mutation on chromosome 2 of the mouse. Mamm Genome 4: 481–484.
13. AlfredJB, RanceK, TaylorBA, PhillipsSJ, AbbottCM, et al. (1997) Mapping in the region of Danforth's short tail and the localization of tail length modifiers. Genome Res 7: 108–117.
14. SembaK, ArakiK, LiZ, MatsumotoK, SuzukiM, et al. (2006) A novel murine gene, Sickle tail, linked to the Danforth's short tail locus, is required for normal development of the intervertebral disc. Genetics 172: 445–456.
15. MaatmanR, ZachgoJ, GosslerA (1997) The Danforth's short tail mutation acts cell autonomously in notochord cells and ventral hindgut endoderm. Development 124: 4019–4028.
16. ArakiK, ArakiM, YamamuraK (2002) Site-directed integration of the cre gene mediated by Cre recombinase using a combination of mutant lox sites. Nucleic Acids Res 30: e103.
17. ZhaoG, LiZ, ArakiK, HarunaK, YamaguchiK, et al. (2008) Inconsistency between hepatic expression and serum concentration of transthyretin in mice humanized at the transthyretin locus. Genes Cells 13: 1257–1268.
18. ArakiK, OkadaY, ArakiM, YamamuraK (2010) Comparative analysis of right element mutant lox sites on recombination efficiency in embryonic stem cells. BMC Biotechnol 10: 29.
19. KawaguchiY, CooperB, GannonM, RayM, MacDonaldRJ, et al. (2002) The role of the transcriptional regulator Ptf1a in converting intestinal to pancreatic progenitors. Nat Genet 32: 128–134.
20. SavoryJGBN, PierreV, RijliFM, De RepentignyY, KotharyR, LohnesD (2009) Cdx2 regulation of posterior development through non-Hox targets. Development 136: 4099–4110.
21. TakadaS, StarkKL, SheaMJ, VassilevaG, McMahonJA, et al. (1994) Wnt-3a regulates somite and tailbud formation in the mouse embryo. Genes Dev 8: 174–189.
22. GrecoTL, TakadaS, NewhouseMM, McMahonJA, McMahonAP, et al. (1996) Analysis of the vestigial tail mutation demonstrates that Wnt-3a gene dosage regulates mouse axial development. Genes Dev 10: 313–324.
23. ChawengsaksophakK, JamesR, HammondVE, KontgenF, BeckF (1997) Homeosis and intestinal tumours in Cdx2 mutant mice. Nature 386: 84–87.
24. MeislerMH (1997) Mutation watch: mouse brachyury (T), the T-box gene family, and human disease. Mamm Genome 8: 799–800.
25. Abu-AbedS, DolleP, MetzgerD, BeckettB, ChambonP, et al. (2001) The retinoic acid-metabolizing enzyme, CYP26A1, is essential for normal hindbrain patterning, vertebral identity, and development of posterior structures. Genes Dev 15: 226–240.
26. SakaiY, MenoC, FujiiH, NishinoJ, ShiratoriH, et al. (2001) The retinoic acid-inactivating enzyme CYP26 is essential for establishing an uneven distribution of retinoic acid along the anterio-posterior axis within the mouse embryo. Genes Dev 15: 213–225.
27. GaoN, WhiteP, KaestnerKH (2009) Establishment of intestinal identity and epithelial-mesenchymal signaling by Cdx2. Dev Cell 16: 588–599.
28. BaillieGJ, van de LagemaatLN, BaustC, MagerDL (2004) Multiple groups of endogenous betaretroviruses in mice, rats, and other mammals. J Virol 78: 5784–5798.
29. PennisiE (2007) Evolution. Jumping genes hop into the evolutionary limelight. Science 317: 894–895.
30. KazazianHHJr (2004) Mobile elements: drivers of genome evolution. Science 303: 1626–1632.
31. MaksakovaIA, RomanishMT, GagnierL, DunnCA, van de LagemaatLN, et al. (2006) Retroviral elements and their hosts: insertional mutagenesis in the mouse germ line. PLoS Genet 2: e2 doi:10.1371/journal.pgen.0020002
32. KanoH, KurahashiH, TodaT (2007) Genetically regulated epigenetic transcriptional activation of retrotransposon insertion confers mouse dactylaplasia phenotype. Proc Natl Acad Sci U S A 104: 19034–19039.
33. KrappA, KnoflerM, LedermannB, BurkiK, BerneyC, et al. (1998) The bHLH protein PTF1-p48 is essential for the formation of the exocrine and the correct spatial organization of the endocrine pancreas. Genes Dev 12: 3752–3763.
34. SellickGS, BarkerKT, Stolte-DijkstraI, FleischmannC, ColemanRJ, et al. (2004) Mutations in PTF1A cause pancreatic and cerebellar agenesis. Nat Genet 36: 1301–1305.
35. HoshinoM, NakamuraS, MoriK, KawauchiT, TeraoM, et al. (2005) Ptf1a, a bHLH transcriptional gene, defines GABAergic neuronal fates in cerebellum. Neuron 47: 201–213.
36. ChesleyP (1935) Development of the short-tailed mutant in the house mouse. J Exp Zool 70: 429–459.
37. GrecoTL, TakadaS, NewhouseMM, McMahonJA, McMahonAP, et al. (1996) Analysis of the vestigial tail mutation demonstrates that Wnt-3a gene dosage regulates mouse axial development. Genes Dev 10: 313–324.
38. YoshikawaY, FujimoriT, McMahonAP, TakadaS (1997) Evidence that absence of Wnt-3a signaling promotes neuralization instead of paraxial mesoderm development in the mouse. Dev Biol 183: 234–242.
39. HoriK, Cholewa-WaclawJ, NakadaY, GlasgowSM, MasuiT, et al. (2008) A nonclassical bHLH Rbpj transcription factor complex is required for specification of GABAergic neurons independent of Notch signaling. Genes Dev 22: 166–178.
40. BeresTM, MasuiT, SwiftGH, ShiL, HenkeRM, et al. (2006) PTF1 is an organ-specific and Notch-independent basic helix-loop-helix complex containing the mammalian Suppressor of Hairless (RBP-J) or its paralogue, RBP-L. Mol Cell Biol 26: 117–130.
41. ArnoldSJ, StappertJ, BauerA, KispertA, HerrmannBG, et al. (2000) Brachyury is a target gene of the Wnt/beta-catenin signaling pathway. Mech Dev 91: 249–258.
42. SaegusaM, HashimuraM, KuwataT, HamanoM, WaniY, et al. (2007) A functional role of Cdx2 in beta-catenin signaling during transdifferentiation in endometrial carcinomas. Carcinogenesis 28: 1885–1892.
43. GuoRJ, HuangE, EzakiT, PatelN, SinclairK, et al. (2004) Cdx1 inhibits human colon cancer cell proliferation by reducing beta-catenin/T-cell factor transcriptional activity. J Biol Chem 279: 36865–36875.
44. GuoRJ, FunakoshiS, LeeHH, KongJ, LynchJP (2010) The intestine-specific transcription factor Cdx2 inhibits beta-catenin/TCF transcriptional activity by disrupting the beta-catenin-TCF protein complex. Carcinogenesis 31: 159–166.
45. KorinekV, BarkerN, MorinPJ, van WichenD, de WegerR, et al. (1997) Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC−/− colon carcinoma. Science 275: 1784–1787.
46. MorinPJ, SparksAB, KorinekV, BarkerN, CleversH, et al. (1997) Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science 275: 1787–1790.
47. RubinfeldB, RobbinsP, El-GamilM, AlbertI, PorfiriE, et al. (1997) Stabilization of beta-catenin by genetic defects in melanoma cell lines. Science 275: 1790–1792.
48. CurrarinoG, ColnD, VottelerT (1981) Triad of anorectal, sacral, and presacral anomalies. AJR Am J Roentgenol 137: 395–398.
49. RossAJ, Ruiz-PerezV, WangY, HaganDM, SchererS, et al. (1998) A homeobox gene, HLXB9, is the major locus for dominantly inherited sacral agenesis. Nat Genet 20: 358–361.
50. BelloniE, MartuccielloG, VerderioD, PontiE, SeriM, et al. (2000) Involvement of the HLXB9 homeobox gene in Currarino syndrome. Am J Hum Genet 66: 312–319.
51. HarrisonKA, DrueyKM, DeguchiY, TuscanoJM, KehrlJH (1994) A novel human homeobox gene distantly related to proboscipedia is expressed in lymphoid and pancreatic tissues. J Biol Chem 269: 19968–19975.
52. HarrisonKA, ThalerJ, PfaffSL, GuH, KehrlJH (1999) Pancreas dorsal lobe agenesis and abnormal islets of Langerhans in Hlxb9-deficient mice. Nat Genet 23: 71–75.
53. LiH, ArberS, JessellTM, EdlundH (1999) Selective agenesis of the dorsal pancreas in mice lacking homeobox gene Hlxb9. Nat Genet 23: 67–70.
54. ThompsonN, GesinaE, ScheinertP, BucherP, Grapin-BottonA (2012) RNA profiling and chromatin immunoprecipitation-sequencing reveal that PTF1a stabilizes pancreas progenitor identity via the control of MNX1/HLXB9 and a network of other transcription factors. Mol Cell Biol 32: 1189–1199.
55. ThomasKR, CapecchiMR (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51: 503–512.
56. YagiT, TokunagaT, FurutaY, NadaS, YoshidaM, et al. (1993) A novel ES cell line, TT2, with high germline-differentiating potency. Anal Biochem 214: 70–76.
57. TaniwakiT, HarunaK, NakamuraH, SekimotoT, OikeY, et al. (2005) Characterization of an exchangeable gene trap using pU-17 carrying a stop codon-beta geo cassette. Dev Growth Differ 47: 163–172.
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2013 Číslo 2
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Complex Inheritance of Melanoma and Pigmentation of Coat and Skin in Grey Horses
- Coordination of Chromatid Separation and Spindle Elongation by Antagonistic Activities of Mitotic and S-Phase CDKs
- Autophagy Induction Is a Tor- and Tp53-Independent Cell Survival Response in a Zebrafish Model of Disrupted Ribosome Biogenesis
- Assembly of the Auditory Circuitry by a Genetic Network in the Mouse Brainstem