title: Mapping cellular processes in the mesenchyme during palatal development in the absence of Tbx1 reveals complex proliferation changes and perturbed cell packing and polarity.
abstract: The 22q11 deletion syndromes represent a spectrum of overlapping conditions including cardiac defects and craniofacial malformations. Amongst the craniofacial anomalies that are seen, cleft of the secondary palate is a common feature. Haploinsufficiency of TBX1 is believed to be a major contributor toward many of the developmental structural anomalies that occur in these syndromes, and targeted deletion of Tbx1 in the mouse reproduces many of these malformations, including cleft palate. However, the cellular basis of this defect is only poorly understood. Here, palatal development in the absence of Tbx1 has been analysed, focusing on cellular properties within the whole mesenchymal volume of the palatal shelves. Novel image analyses and data presentation tools were applied to quantify cell proliferation rates, including regions of elevated as well as reduced proliferation, and cell packing in the mesenchyme. Also, cell orientations (nucleus-Golgi axis) were mapped as a potential marker of directional cell movement. Proliferation differed only subtly between wild-type and mutant until embryonic day (E)15.5 when proliferation in the mutant was significantly lower. Tbx1(-/-) palatal shelves had slightly different cell packing than wild-type, somewhat lower before elevation and higher at E15.5 when the wild-type palate has elevated and fused. Cell orientation is biased towards the shelf distal edge in the mid-palate of wild-type embryos but is essentially random in the Tbx1(-/-) mutant shelves, suggesting that polarised processes such as directed cell rearrangement might be causal for the cleft phenotype. The implications of these findings in the context of further understanding Tbx1 function during palatogenesis and of these methods for the more general analysis of genotype-phenotype functional relationships are discussed.
authors.affiliation: Department of Craniofacial Development and Stem Cell Biology, King's College London, London, UK.
authors.forename: Lara J
authors.initials: LJ
authors.lastname: Brock
authors.affiliation: Department of Craniofacial Development and Stem Cell Biology, King's College London, London, UK.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Economou
authors.affiliation: Department of Craniofacial Development and Stem Cell Biology, King's College London, London, UK.
authors.forename: Martyn T
authors.initials: MT
authors.lastname: Cobourne
authors.affiliation: Department of Craniofacial Development and Stem Cell Biology, King's College London, London, UK.
authors.forename: Jeremy B A
authors.identifier: http://orcid.org/0000-0002-6102-2620
authors.initials: JB
authors.lastname: Green
mesh_terms: D058165:22q11 Deletion Syndrome / Q000196:embryology* / Q000235:genetics; D000818:Animals; D016764:Cell Polarity / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D002972:Cleft Palate / Q000235:genetics*; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D007091:Image Processing, Computer-Assisted; D008648:Mesoderm / Q000196:embryology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010159:Palate / Q000196:embryology*; D020825:T-Box Domain Proteins / Q000172:deficiency* / Q000235:genetics
chemical_list: D020825:T-Box Domain Proteins; C491573:Tbx1 protein, mouse
keywords: Tbx1; cell orientation; cell packing; cell proliferation; cleft palate; mesenchyme