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Histochemistry and Cell Biology

, Volume 136, Issue 6, pp 663–675 | Cite as

Shh signaling is essential for rugae morphogenesis in mice

  • Jong-Min Lee
  • Seita Miyazawa
  • Jeong-Oh Shin
  • Hyuk-Jae Kwon
  • Dae-Woon Kang
  • Byung-Jai Choi
  • Jae-Ho Lee
  • Shigeru Kondo
  • Sung-Won Cho
  • Han-Sung JungEmail author
Original Paper

Abstract

Palatal ridges, or rugae palatinae, are corrugated structures observed in the hard palate region. They are found in most mammalian species, but their number and arrangement are species-specific. Nine palatal rugae are found in the mouse secondary palate. Previous studies have shown that epithelial Shh signaling in the palatal ridge plays an important role during rugae development. Moreover, Wnt family members, including LEF1, play a functional role in orofacial morphogenesis. To explore the function of Shh during rugae development, we utilized the maternal transfer of 5E1 (anti-Shh antibody) to mouse embryos. 5E1 induced abnormal rugae patterning characterized by a spotted shape of palatal ridge rather than a stripe. The expression patterns of Shh and Shh-related genes, Sostdc1, Lef1 and Ptch1, were disrupted following 5E1 injection. Moreover, rugae-specific cell proliferation and inter-rugae-specific apoptosis were affected by inhibition of Shh signaling. We hypothesize that the altered gene expression patterns and the change in molecular events caused by the inhibition of Shh signaling may have induced abnormal rugae patterning. Furthermore, we propose a reaction–diffusion model generated by Wnt, Shh and Sostdc1 signaling. In this study, we show that Sostdc1, a secreted inhibitor of the Wnt pathway, is a downstream target of Shh and hypothesize that the interaction of Wnt, Shh and Sostdc1 is a pivotal mechanism controlling the spatial patterning of palatal rugae.

Keywords

Rugae patterning Wnt Shh Sostdc1 Reaction–diffusion 

Notes

Acknowledgment

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (R13-2003-013-05001-0).

Supplementary material

418_2011_870_MOESM1_ESM.tif (6 mb)
Supplementary figure 1. Gli1, Bmp4 and Lrp4 expression patterns in rugae following 5E1 injection. (a, a′, d, d′) Gli1 expression is reduced in the palatal ridge 24 h after 5E1 delivery. (b, b′, e, e′, c, c′, f, f′) Bmp4 and Lrp4 expression are not altered 24 h after 5E1 injection. (g, g′, j, j′) Gli1 expression is clearly reduced in the palatal ridge 48 h after 5E1 injection. (i, i′, l, l′) Lrp4 expression is not altered 48 h after 5E1 injection. (h, h′, k, k′) In the control group, Bmp4 is expressed along the first three rugae lane. However, Bmp4 expression was detected in only in the first rugae following 5E1 treatment. Scale bar; a-l: 500 μm, a′–l′ : 100 μm. (TIFF 6138 kb)
418_2011_870_MOESM2_ESM.tif (305 kb)
Supplementary figure 2. Alteration of Wnt/β-catenin signaling levels following 5E1 injection. Gene expression levels are examined by microarray. The Wnt/β-catenin downstream genes, Axin2, Dkk1 and Gata3, expression levels are up-regulated after 5E1 treatment. (TIFF 305 kb)

Supplementary movie 1. An additional row of cells is inserted into the anterior part of the #8 ruga. Newly formed rugae appeared following the reaction–diffusion model that consists of activator, mediator and inhibitor. The lateral extension is simulated by the addition of a new column of cells to the edge of the field between #3 and #4 rugae formations. (MP4 1272 kb)

References

  1. Ahn Y, Sanderson BW, Klein OD, Krumlauf R (2010) Inhibition of Wnt signaling by Wise (Sostdc1) and negative feedback from Shh controls tooth number and patterning. Development 137:3221–3231PubMedCrossRefGoogle Scholar
  2. Andrade Filho PA, Letra A, Cramer A, Prasad JL, Garlet GP, Vieira AR, Ferris RL, Menezes R (2011) Insights from studies with oral cleft genes suggest associations between WNT-pathway genes and risk of oral cancer. J Dent Res 90:740–746PubMedCrossRefGoogle Scholar
  3. Bitgood MJ, McMahon AP (1995) Hedgehog and Bmp genes are coexpressed at many diverse sites of cell–cell interaction in the mouse embryo. Dev Biol 172:126–138PubMedCrossRefGoogle Scholar
  4. Blanton SH, Bertin T, Serna ME, Stal S, Mulliken JB, Hecht JT (2004) Association of chromosomal regions 3p21.2, 10p13, and 16p13.3 with nonsyndromic cleft lip and palate. Am J Med Genet A 125:23–27CrossRefGoogle Scholar
  5. Bosanac I, Maun HR, Scales SJ, Wen X, Lingel A, Bazan JF, de Sauvage FJ, Hymowitz SG, Lazarus RA (2009) The structure of SHH in complex with HHIP reveals a recognition role for the Shh pseudo active site in signaling. Nat Struct Mol Biol 16:691–697PubMedCrossRefGoogle Scholar
  6. Brown NL, Knott L, Halligan E, Yarram SJ, Mansell JP, Sandy JR (2003) Microarray analysis of murine palatogenesis: temporal expression of genes during normal palate development. Dev Growth Differ 45:153–165PubMedCrossRefGoogle Scholar
  7. Cai J, Cho SW, Kim JY, Lee MJ, Cha YG, Jung HS (2007) Patterning the size and number of tooth and its cusps. Dev Biol 304:499–507 Google Scholar
  8. Chen Y, Struhl G (1996) Dual roles for patched in sequestering and transducing Hedgehog. Cell 87:553–563PubMedCrossRefGoogle Scholar
  9. Chiang C, Litingtung Y, Lee E, Young KE, Corden JL, Wesphal H, Beachy P (1996) Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature 383:407–413PubMedCrossRefGoogle Scholar
  10. Cho SW, Kwak S, Woolley TE, Lee MJ, Kim EJ, Baker RE, Kim HJ, Shin JS, Tickle C, Maini PK, Jung HS (2011) Interactions between Shh, Sostdc1 and Wnt signaling and a new feedback loop for spatial patterning of the teeth. Development 138:1807–1816PubMedCrossRefGoogle Scholar
  11. Eisentraut M (1979) Das Gaumenfaltenmuster der Säugetiere und seine Bedeutung für Stammesgeschichtliche und taxonomische Untersuchungen. Bonner Zool Monogr 8:211–214Google Scholar
  12. Ferguson MW (1987) Palate development: mechanisms and malformations. Ir J Med Sci 156:309–315PubMedCrossRefGoogle Scholar
  13. Goodrich LV, Johnson RL, Milenkovic L, McMahon JA, Scott MP (1996) Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog. Genes Dev 10:301–312PubMedCrossRefGoogle Scholar
  14. Goodrich LV, Milenkovic L, Higgins KM, Scott MP (1997) Altered neural cell fates and medulloblastoma in mouse patched mutants. Science 277:1109–1113PubMedCrossRefGoogle Scholar
  15. Gritli-Linde A (2007) Molecular control of secondary palate development. Dev Biol 301:309–326PubMedCrossRefGoogle Scholar
  16. Grote D, Boualia SK, Souabni A, Merkel C, Chi X, Costantini F, Carroll T, Bouchard M (2008) Gata3 acts downstream of beta-catenin signaling to prevent ectopic metanephric kidney induction. PLoS Genet 4(12):e1000316PubMedCrossRefGoogle Scholar
  17. Ichikawa H, Matsuo S, Silos-Santiago I, Jacquin MF, Sugimoto T (2001) Developmental dependency of Merkel endings on trks in the palate. Brain Res Mol Brain Res 88:171–175PubMedCrossRefGoogle Scholar
  18. Jernvall J, Thesleff I (2000) Reiterative signaling and patterning during mammalian tooth morphogenesis. Mech Dev 92:19–29 Google Scholar
  19. Johnson EB, Hammer RE, Herz J (2005) Abnormal development of the apical ectodermal ridge and polysyndactyly in Megf7-deficient mice. Hum Mol Genet 14:3523–3538PubMedCrossRefGoogle Scholar
  20. Juriloff DM, Harris MJ, McMahon AP, Carroll TJ, Lidral AC (2006) Wnt9b is the mutated gene involved in multifactorial nonsyndromic cleft lip with or without cleft palate in A/WySn mice, as confirmed by a genetic complementation test. Birth Defects Res A Clin Mol Teratol 76:574–579PubMedCrossRefGoogle Scholar
  21. Kido MA, Muroya H, Yamaza T, Terada Y, Tanaka T (2003) Vanilloid receptor expression in the rat tongue and palate. J Dent Res 82:393–397PubMedCrossRefGoogle Scholar
  22. Kratochwil K, Galceran J, Tontsch S, Roth W, Grosschedl R (2002) FGF4, a direct target of LEF1 and Wnt signaling, can rescue the arrest of tooth organogenesis in Lef1–/–mice. Genes Dev 16:3173–3185PubMedCrossRefGoogle Scholar
  23. Lan Y, Ryan RC, Zhang Z, Bullard SA, Bush JO, Maltby KM, Lidral AC, Jiang R (2006) Expression of Wnt9b and activation of canonical Wnt signaling during midfacial morphogenesis in mice. Dev Dyn 235:1448–1454PubMedCrossRefGoogle Scholar
  24. Lee JM, Kim JY, Cho KW, Lee MJ, Cho SW, Kwak S, Cai J, Jung HS (2008) Wnt11/Fgfr1b cross-talk modulates the fate of cells in palate development. Dev Biol 314:341–350PubMedCrossRefGoogle Scholar
  25. Letra A, Menezes R, Granjeiro JM, Vieira AR (2009) AXIN2 and CDH1 polymorphisms, tooth agenesis, and oral clefts. Birth Defects Res A Clin Mol Teratol 85:169–173PubMedCrossRefGoogle Scholar
  26. Li Y, Pawlik B, Elcioglu N, Aglan M, Kayserili H, Yigit G, Percin F, Goodman F, Nürnberg G, Cenani A, Urquhart J, Chung BD, Ismail S, Amr K, Aslanger AD, Becker C, Netzer C, Scambler P, Eyaid W, Hamamy H, Clayton-Smith J, Hennekam R, Nürnberg P, Herz J, Temtamy SA, Wollnik B (2010) LRP4 mutations alter Wnt/beta-catenin signaling and cause limb and kidney malformations in Cenani-Lenz syndrome. Am J Hum Genet 86:696–706PubMedCrossRefGoogle Scholar
  27. Liu F, Chu EY, Watt B, Zhang Y, Gallant NM, Andl T, Yang SH, Lu MM, Piccolo S, Schmidt-Ullrich R, Taketo MM, Morrisey EE, Atit R, Dlugosz AA, Millar SE (2008) Wnt/beta-catenin signaling directs multiple stages of tooth morphogenesis. Dev Biol 313:210–224PubMedCrossRefGoogle Scholar
  28. Mitsui C, Iwanaga T, Yoshida S, Kawasaki T (2000) Immunohistochemical demonstration of nerve terminals in the whole hard palate of rats by use of an antiserum against protein gene product 9.5 (PGP 9.5). Arch Histol Cytol 63:401–410PubMedCrossRefGoogle Scholar
  29. Mohamed OA, Clarke HJ, Dufort D (2004) Beta-catenin signaling marks the prospective site of primitive streak formation in the mouse embryo. Dev Dyn 231:416–424PubMedCrossRefGoogle Scholar
  30. Monaghan AP, Kioschis P, Wu W, Zuniga A, Bock D, Poustka A, Delius H, Niehrs C (1999) Dickkopf genes are co-ordinately expressed in mesodermal lineages. Mech Dev 87:45–56PubMedCrossRefGoogle Scholar
  31. Nawshad A, Hay ED (2003) TGF beta 3 signaling activates transcription of the LEF1 gene to induce epithelial–mesenchymal transformation during mouse palate development. J Cell Biol 163:1291–1301PubMedCrossRefGoogle Scholar
  32. Nunzi MG, Pisarek A, Mugnaini E (2004) Merkel cells, corpuscular nerve endings and free nerve endings in the mouse palatine mucosa express three subtypes of vesicular glutamate transporters. J Neurocytol 33:359–376PubMedCrossRefGoogle Scholar
  33. Ohazama A, Johnson EB, Ota MS, Choi HY, Porntaveetus T, Oommen S, Itoh N, Eto K, Gritli-Linde A, Herz J, Sharpe PT (2008) Lrp4 modulates extracellular integration of cell signaling pathways in development. PLoS One 3:e4092PubMedCrossRefGoogle Scholar
  34. Oosterwegel M, Timmerman J, Leiden J, Clevers H (1992) Expression of GATA-3 during lymphocyte differentiation and mouse embryogenesis. Dev Immunol 3:1–11PubMedCrossRefGoogle Scholar
  35. Pantalacci S, Prochazka J, Martin A, Rothova M, Lambert A, Bernard L, Charles C, Viriot L, Peterkova R, Laudet V (2008) Patterning of palatal rugae through sequential addition reveals an anterior/posterior boundary in palatal development. BMC Dev Biol 8:116–133PubMedCrossRefGoogle Scholar
  36. Peterková R, Klepácek I, Peterka M (1987) Prenatal development of rugae palatinae in mice: scanning electron microscopic and histologic studies. J Craniofac Genet Dev Biol 7:169–189PubMedGoogle Scholar
  37. Porntaveetus T, Oommen S, Sharpe PT, Ohazama A (2010) Expression of Fgf signalling pathway related genes during palatal rugae development in the mouse. Gene Expr Patterns 10:193–198PubMedCrossRefGoogle Scholar
  38. Rice R, Spencer-Dene B, Connor EC, Gritli-Linde A, McMahon AP, Dickson C, Thesleff I, Rice DP (2004) Disruption of Fgf10/Fgfr2b-coordinated epithelial–mesenchymal interactions causes cleft palate. J Clin Invest 113:1692–1700PubMedGoogle Scholar
  39. Sakamoto MK, Nakamura K, Handa J, Kihara T, Tanimura T (1989) Morphogenesis of the secondary palate in mouse embryos with special reference to the development of rugae. Anat Rec 223:299–310PubMedCrossRefGoogle Scholar
  40. Salazar-Ciudad I, Jernvall J (2010) A computational model of teeth and the developmental origins of morphological variation. Nature 464:583–586PubMedCrossRefGoogle Scholar
  41. Sohn WJ, Yamamoto H, Shin HI, Ryoo ZY, Lee S, Bae YC, Jung HS, Kim JY (2011) Importance of region-specific epithelial rearrangements in mouse rugae development. Cell Tissue Res 344:271–277PubMedCrossRefGoogle Scholar
  42. Tian E, Zhan F, Walker R, Rasmussen E, Ma Y, Barlogie B, Shaughnessy JD Jr (2003) The role of the Wnt-signaling antagonist DKK1 in the development of osteolytic lesions in multiple myeloma. N Engl J Med 349:2483–2494PubMedCrossRefGoogle Scholar
  43. Tyler MS, Koch WE (1977) In vitro development of palatal tissues from embryonic mice. II. Tissue isolation and recombination studies. J Embryol Exp Morphol 38:19–36PubMedGoogle Scholar
  44. Valdivia LE, Young RM, Hawkins TA, Stickney HL, Cavodeassi F, Schwarz Q, Pullin LM, Villegas R, Moro E, Argenton F, Allende ML, Wilson SW (2011) Lef1-dependent Wnt/β-catenin signalling drives the proliferative engine that maintains tissue homeostasis during lateral line development. Development 138:3931–3941PubMedCrossRefGoogle Scholar
  45. Wang LC, Liu ZY, Gambardella L, Delacour A, Shapiro R, Yang J, Sizing I, Rayhorn P, Garber EA, Benjamin CD, Williams KP, Taylor FR, Barrandon Y, Ling L, Burkly LC (2000) Regular articles: conditional disruption of hedgehog signaling pathway defines its critical role in hair development and regeneration. J Invest Dermatol 114:901–908PubMedCrossRefGoogle Scholar
  46. Welsh IC, O’Brien TP (2009) Signaling integration in the rugae growth zone directs sequential SHH signaling center formation during the rostral outgrowth of the palate. Dev Biol 336:53–67PubMedCrossRefGoogle Scholar
  47. Wu X, Daniels G, Shapiro E, Xu K, Huang H, Li Y, Logan S, Greco MA, Peng Y, Monaco ME, Melamed J, Lepor H, Grishina I, Lee P (2011) LEF1 identifies androgen-independent epithelium in the developing prostate. Mol Endocrinol 25:1018–1026PubMedCrossRefGoogle Scholar
  48. Zhang Z, Song Y, Zhao X, Zhang X, Fermin C, Chen Y (2002) Rescue of cleft palate in Msx1-deficient mice by transgenic Bmp4 reveals a network of BMP and Shh signaling in the regulation of mammalian palatogenesis. Development 129:4135–4146PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Jong-Min Lee
    • 1
  • Seita Miyazawa
    • 2
  • Jeong-Oh Shin
    • 1
  • Hyuk-Jae Kwon
    • 1
  • Dae-Woon Kang
    • 3
  • Byung-Jai Choi
    • 3
  • Jae-Ho Lee
    • 3
  • Shigeru Kondo
    • 2
  • Sung-Won Cho
    • 1
  • Han-Sung Jung
    • 1
    Email author
  1. 1.Division in Anatomy and Developmental Biology, Department of Oral Biology, Research Center for Orofacial Hard Tissue Regeneration, Brain Korea 21 Project, Oral Science Research Center, College of Dentistry, Yonsei Center of BiotechnologyYonsei UniversitySeoulKorea
  2. 2.Graduate School of Frontier BiosciencesOsaka UniversityOsakaJapan
  3. 3.Department of Pediatric Dentistry, Oral Science Research CenterCollege of Dentistry, Yonsei UniversitySeoulKorea

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