title: Matrix metalloproteinase-25 has a functional role in mouse secondary palate development and is a downstream target of TGF-β3.
abstract: BACKGROUND
  Development of the secondary palate (SP) is a complex event and abnormalities during SP development can lead to cleft palate, one of the most common birth disorders. Matrix metalloproteinases (MMPs) are required for proper SP development, although a functional role for any one MMP in SP development remains unknown. MMP-25 may have a functional role in SP formation as genetic scans of the DNA of human cleft palate patients indicate a common mutation at a region upstream of the MMP-25 gene. We report on the gene expression profile of MMP-25 in the developing mouse SP and identify its functional role in mouse SP development.
  
  
  RESULTS
  MMP-25 mRNA and protein are found at all SP developmental stages in mice, with the highest expression at embryonic day (E) 13.5. Immunohistochemistry and in situ hybridization localize MMP-25 protein and mRNA, respectively, to the apical palate shelf epithelial cells and apical mesenchyme. MMP-25 knockdown with siRNA in palatal cultures results in a significant decrease in palate shelf fusion and persistence of the medial edge epithelium. MMP-25 mRNA and protein levels significantly decrease when cultured palate shelves are incubated in growth medium with 5 μg/mL of a TGF-β3-neutralizing antibody.
  
  
  CONCLUSIONS
  Our findings indicate: (i) MMP-25 gene expression is highest at E12.5 and E13.5, which corresponds with increasing palate shelf growth downward alongside the tongue; (ii) MMP-25 protein and mRNA expression predominantly localize in the apical epithelium of the palate shelves, but are also found in apical areas of the mesenchyme; (iii) knockdown of MMP-25 mRNA expression impairs palate shelf fusion and results in significant medial edge epithelium remaining in contacted areas; and (iv) bio-neutralization of TGF-β3 significantly decreases MMP-25 gene expression. These data suggest a functional role for MMP-25 in mouse SP development and are the first to identify a role for a single MMP in mouse SP development.
authors.affiliation: Laboratory of Molecular Biology, College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5C9, Canada.
authors.forename: Graham D
authors.initials: GD
authors.lastname: Brown
authors.forename: Adil J
authors.initials: AJ
authors.lastname: Nazarali
mesh_terms: D000818:Animals; D005260:Female; D058851:GPI-Linked Proteins; D066298:In Vitro Techniques; D053504:Matrix Metalloproteinases, Membrane-Associated / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D010159:Palate / Q000196:embryology* / Q000378:metabolism; D053782:Transforming Growth Factor beta3 / Q000378:metabolism*
chemical_list: D058851:GPI-Linked Proteins; D053782:Transforming Growth Factor beta3; D053504:Matrix Metalloproteinases, Membrane-Associated; C403928:matrix metalloproteinase 25
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