title: Altered mandibular development precedes the time of palate closure in mice homozygous for disproportionate micromelia: an oral clefting model supporting the Pierre-Robin sequence.
abstract: BACKGROUND
  Development of the human craniofacial anatomy involves a number of interrelated, genetically controlled components. The complexity of the interactions between these components suggests that interference with the spaciotemporal interaction of the expanding tongue and elongating Meckel's cartilage correlates with the appearance of cleft palate. Mice homozygous for the semi-dominant Col2a1 mutation Disproportionate micromelia (Dmm), presenting at birth with both cleft palate and micrognathia, provide the opportunity to test the hypothesis that mandibular growth retardation coincides with formation of the secondary palate as predicted from our understanding of the Pierre Robin sequence. The present study was conducted in embryonic day 14 (E14) mice, 1 day before palate closure, to describe the relationship between growth of the lower jaw/tongue complex versus genotype of the embryo.
  
  
  METHODS
  Whole heads, isolated from E14.25, E14.5 and E14.75 wild-type and homozygous mutant embryos, were fixed in Bouin's solution, embedded in paraffin, and serially sectioned. Mid-sagittal sections, stained with toluidine blue, were used to estimate growth of both tongue and lower jaw (Meckel's cartilage length) during a 12-hr period preceding palate closure.
  
  
  RESULTS
  In control embryos, the largest increase in Meckel's cartilage length occurred between E14.5 and E14.75. Compared to control, the mean Meckel's cartilage length in the mutant was similar at E14.25, but was significantly less at E14.5 and E14.75. Absolute tongue size in control embryos increased linearly during this period of E14.25 to E14.75. Relative to the rapidly growing Meckel's cartilage, however, relative tongue size in control embryos actually decreased over time. Absolute tongue size in the mutant was not significantly different from that of control at any of the embryonic stages examined, however, relative tongue size in the mutant was significantly greater at E14.75 compared to control.
  
  
  CONCLUSIONS
  Mandibular growth retardation, coupled with relative macroglossia in E14 Dmm/Dmm mice, suggests that the concerted development of the palate and lower jaw complex in the mutant is aberrant. Detection of micrognathia and pseudomacroglossia in homozygotes, before the time of palate closure, supports the hypothesis that a relationship exists between growth retardation of Meckel's cartilage and malformation of the secondary palate, as predicted by the Pierre-Robin sequence.
authors.affiliation: Department of Zoology, Brigham Young University, Provo, Utah 84602, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Ricks
authors.forename: Victoria M
authors.initials: VM
authors.lastname: Ryder
authors.forename: Laura C
authors.initials: LC
authors.lastname: Bridgewater
authors.forename: Bruce
authors.initials: B
authors.lastname: Schaalje
authors.forename: Robert E
authors.initials: RE
authors.lastname: Seegmiller
mesh_terms: D000818:Animals; D002972:Cleft Palate / Q000196:embryology* / Q000235:genetics; D024043:Collagen Type II / Q000235:genetics*; D006720:Homozygote; D008334:Mandible / Q000002:abnormalities / Q000196:embryology*; D051379:Mice; D008809:Mice, Inbred C3H; D008844:Micrognathism / Q000196:embryology / Q000235:genetics; D010159:Palate / Q000002:abnormalities / Q000196:embryology*; D010855:Pierre Robin Syndrome / Q000196:embryology* / Q000235:genetics; D017384:Sequence Deletion
chemical_list: C447413:Col2a1 protein, mouse; D024043:Collagen Type II