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Informationen aus Orthodontie & Kieferorthopädie 2017; 49(02): 148-152
DOI: 10.1055/s-0043-109044
Übersichtsartikel
© Georg Thieme Verlag KG Stuttgart · New York

Ein In-vitro-Mausmodell für die Palatogenese und die Entstehung von Gaumenspalten

A Mouse In Vitro Model for Palatogenesis and Cleft Palate
Johannes W Von den Hoff
1   Department of Orthodontics and Craniofacial Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Nijmegen, The Netherlands
,
Laury A Roa Fuentes
1   Department of Orthodontics and Craniofacial Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Nijmegen, The Netherlands
,
Marjon Bloemen
2   Department of Orthodontics and Craniofacial Biology, Radboud University Medical Center, Nijmegen, The Netherlands
,
Anne Marie Kuijpers-Jagtman
2   Department of Orthodontics and Craniofacial Biology, Radboud University Medical Center, Nijmegen, The Netherlands
› Author Affiliations
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Publication History

Publication Date:
20 July 2017 (online)

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Abstract

The mouse is an excellent model to study palatogenesis since, like humans, it is a mammal, it can easily be bred and a wide array of molecular tools are available. Moreover, the isolated embryonic mouse palate can be cultured in vitro, which allows us to study the actual fusion process in detail.

Just prior to the actual fusion at E13, the maxilla with the 2 palatal shelves can be dissected from the embryo head. The maxilla is cultured on a filter paper on top of a stainless-steel grid at the interface of the culture medium and air. After 24 h in culture, the palatal shelves are in contact and the MES is clearly visible. At 48 h, the MES has nearly completely disintegrated with small remnants at the nasal and oral side of the palate. Finally, at 72 h, the palatal shelves are completely fused and osteoid tissue is beginning to form in the lateral areas.

The exact downstream molecular mechanisms that lead to clefting are far from being unraveled. The model for palate fusion presented here offers the possibility to analyze the molecular and cellular mechanisms that lead to clefting as a result of specific genetic and environmental factors. This will pave the way to pharmacological intervention for the prevention of cleft lip and/or palate in susceptible individuals.

Zusammenfassung

Das Mausmodell eignet sich für die Untersuchung der Palatogenese hervorragend, da Mäuse, genau wie Menschen, zu den Säugetieren gehören. Sie lassen sich leicht vermehren und es steht für sie eine Vielzahl an molekularen Werkzeugen zur Verfügung. Außerdem lässt sich der isolierte Gaumen von Mäusen in vitro kultivieren, wodurch detaillierte Untersuchungen des Verschmelzungsprozesses möglich sind.

Kurz vor der eigentlichen Verschmelzung während E13 kann die Maxilla mit den beiden Gaumenfortsätzen aus dem Schädel des Mausembryos entnommen werden. Der Oberkiefer wird anschließend auf einem Edelstahlgitter mit Filterpapier an der Oberfläche eines Kulturmediums angezüchtet. Nach 24 Stunden in Kultur haben die beiden Gaumenfortsätze Kontakt und der MES ist deutlich zu erkennen. Nach 48 Stunden hat sich der MES fast vollständig aufgelöst und es sind nur noch geringe Reste auf der nasalen und der oralen Seite des Gaumens vorhanden. Nach 72 Stunden sind die beiden Gaumenfortsätze schließlich vollständig verschmolzen und in den seitlichen Regionen bildet sich Osteoidgewebe aus.

Die exakte Abfolge der molekularen Mechanismen ist im Einzelnen allerdings noch weitgehend ungeklärt. Das hier vorgestellte Modell für die Verschmelzung der Gaumenfortsätze bietet jedoch die Möglichkeit, die molekularen und zellulären Mechanismen zu untersuchen, die in Folge spezieller genetischer und äußerer Faktoren zur Entstehung von Spalten führen. Auf diese Weise wird es vielleicht einmal möglich werden, die Entstehung von Lippen- bzw. Gaumenspalten bei dafür anfälligen Personen pharmakologisch zu verhindern.

Schlüsselwörter

Gaumenspalte - Embryologie - Maus - In-vitro-Verfahren

Key words

cleft palate - embryology - mouse - in vitro techniques
 

  • Literatur

  • 1 Gritli-Linde A. Molecular control of secondary palate development. Dev Biol 2007; 301: 309-326
    CrossrefPubMedGoogle Scholar
  • 2 Stanier P, Moore GE. Genetics of cleft lip and palate: syndromic genes contribute to the incidence of non-syndromic clefts. Hum Mol Genet 2004; 13 Spec No 1 R73-R81
    CrossrefPubMedGoogle Scholar
  • 3 Dixon MJ, Marazita ML, Beaty TH. et al. Cleft lip and palate: understanding genetic and environmental influences. Nat Rev Genet 2011; 12: 167-178
    CrossrefPubMedGoogle Scholar
  • 4 Vieira AR. Genetic and environmental factors in human cleft lip and palate. Front Oral Biol 2012; 16: 19-31
    PubMedGoogle Scholar
  • 5 Setó-Salvia N, Stanier P. Genetics of cleft lip and/or cleft palate: association with other common anomalies. Eur J Med Genet 2014; 57: 381-393
    CrossrefPubMedGoogle Scholar
  • 6 Jiang R, Bush JO, Lidral AC. Development of the upper lip: morphogenetic and molecular mechanisms. Dev Dyn 2006; 235: 1152-1166
    CrossrefPubMedGoogle Scholar
  • 7 Ferguson MW. Palate development. Development 1988; 103 Suppl: 41-60
    PubMedGoogle Scholar
  • 8 Gato A, Martinez ML, Tudela C. et al. TGF-beta(3)-induced chondroitin sulphate proteoglycan mediates palatal shelf adhesion. Dev Biol 2002; 250: 393-405
    CrossrefPubMedGoogle Scholar
  • 9 Mogass M, Bringas Jr. P, Shuler CF. Characterization of desmosomal component expression during palatogenesis. Int J Dev Biol 2000; 44: 317-322
    PubMedGoogle Scholar
  • 10 Cuervo R, Covarrubias L. Death is the major fate of medial edge epithelial cells and the cause of basal lamina degradation during palatogenesis. Development 2004; 131: 15-24
    CrossrefPubMedGoogle Scholar
  • 11 Martinez-Alvarez C, Tudela C, Perez-Miguelsanz J. et al. Medial edge epithelial cell fate during palatal fusion. Dev Biol 2000; 220: 343-357
    CrossrefPubMedGoogle Scholar
  • 12 Jin JZ, Ding J. Analysis of cell migration, transdifferentiation and apoptosis during mouse secondary palate fusion. Development 2006; 133: 3341-3347
    CrossrefPubMedGoogle Scholar
  • 13 Vaziri Sani F, Hallberg K, Harfe BD. et al. Fate-mapping of the epithelial seam during palatal fusion rules out epithelial-mesenchymal transformation. Dev Biol 2005; 285: 490-495
    CrossrefPubMedGoogle Scholar
  • 14 Moxham BJ. The development of the palate-a brief review. Eur J Anat 2003; 7 (Suppl. 01) 53-74
    PubMedGoogle Scholar
  • 15 Taya Y, O'Kane S, Ferguson MW. Pathogenesis of cleft palate in TGF-beta3 knockout mice. Development 1999; 126: 3869-3879
    PubMedGoogle Scholar
  • 16 Tudela C, Formoso MA, Martínez T. et al. TGF-beta3 is required for the adhesion and intercalation of medial edge epithelial cells during palate fusion. Int J Dev Biol 2002; 46: 333-336
    PubMedGoogle Scholar
  • 17 Dudas M, Nagy A, Laping NJ. et al. Tgf-beta3-induced palatal fusion is mediated by Alk-5/Smad pathway. Dev Biol 2004; 266: 96-108
    CrossrefPubMedGoogle Scholar
  • 18 Sasaki Y, O'Kane S, Dixon J. et al. Temporal and spatial expression of Pax9 and Sonic hedgehog during development of normal mouse palates and cleft palates in TGF-beta3 null embryos. Arch Oral Biol 2007; 52: 260-267
    CrossrefPubMedGoogle Scholar
  • 19 Mima J, Koshino A, Oka K. et al. Regulation of the epithelial adhesion molecule CEACAM1 is important for palate formation. PLoS One 2013; 17 8: e61653
    CrossrefPubMedGoogle Scholar
  • 20 Crisera C, Teng E, Wasson KL. et al. Formation of in vitro murine cleft palate by abrogation of fibroblast growth factor signaling. Plast Reconstr Surg 2008; 121: 218-224
    CrossrefPubMedGoogle Scholar
  • 21 Shimizu N, Aoyama H, Hatakenaka N. et al. An in vitro screening system for characterizing the cleft palate-inducing potential of chemicals and underlying mechanisms. Reprod Toxicol 2001; 15: 665-672
    CrossrefPubMedGoogle Scholar
  • 22 Kang P, Svoboda KK. Nicotine inhibits palatal fusion and modulates nicotinic receptors and the PI-3 kinase pathway in medial edge epithelia. Orthod Craniofac Res 2003; 6: 129-142
    CrossrefPubMedGoogle Scholar
  • 23 Gunnerbeck A, Edstedt Bonamy AK, Wikström AK. et al. Maternal snuff use and smoking and the risk of oral cleft malformations-a population-based cohort study. PLoS One 2014; 9: e84715
    CrossrefPubMedGoogle Scholar
  • 24 Abbott BD, Buckalew AR, Leffler KE. Effects of epidermal growth factor (EGF), transforming growth factor-alpha (TGFalpha), and 2,3,7,8-tetrachlorodibenzo-p-dioxin on fusion of embryonic palates in serum-free organ culture using wild-type, EGF knockout, and TGFalpha knockout mouse strains. Birth Defects Res A Clin Mol Teratol 2005; 73: 447-454
    CrossrefPubMedGoogle Scholar
  • 25 Yao Z, Chen D, Wang A. et al. Folic acid rescue of ATRA-induced cleft palate by restoring the TGF-ß signal and inhibiting apoptosis. J Oral Pathol Med 2011; 40: 433-439
    CrossrefPubMedGoogle Scholar
  • 26 Meng L, Wang X, Torensma R. et al. Lithium inhibits palatal fusion and osteogenic differentiation in palatal shelves in vitro. Arch Oral Biol 2015; 60: 501-507
    CrossrefPubMedGoogle Scholar