title: Isolation of epithelial cells in the developing primary lip and palate.
abstract: Failure of the primary lip and palate to fuse leads to clefts of the lip, a birth defect with an incidence of 1 for every 500 in some races. Epithelial cells lining the facial processes of the primary lip and palate, the lateral and medial nasal processes (LNP and MNP), must first make contact to go through a series of highly regulated and coordinated sequence of events to form the normal midface. As yet, many of the basic mechanisms underlying the fusion events of the epithelial-lined surfaces are not known. This is due in part to the difficulty associated with the isolation of the epithelial cells for further study and analysis. The objective of this study was to test the use of laser capture microdissection to collect clean populations of primary lip and palate epithelial cells destined to fuse. Fusing and nonfusing epithelial cell populations of the MNP and LNP were isolated by laser capture microdissection and assayed for gene expression of Bmp-4, Tgfβ-2, and their type 1 receptors, Alk-3 and Alk-5, respectively. Transcripts of Bmp-4/Alk-3 and Tgfβ-2/Alk-5 were restricted to the epithelial seam of the fusion site, and the epithelium of the prefusion site, in patterns previously reported. Data indicated our ability to isolate clean populations of epithelial and mesenchymal cells around the primary palate fusion site, allowing precise analysis of tissue and site-specific gene expression at high resolution. This study provides the basis of further analysis of the potential molecular players of MNP and LNP fusion and nonfusion of epithelial cells.
authors.affiliation: School of Dentistry, Department of Otolaryngology - Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Shih-Hao
authors.initials: SH
authors.lastname: Huang
authors.forename: Tzy-Wen
authors.initials: TW
authors.lastname: Gong
authors.forename: Siew-Ging
authors.initials: SG
authors.lastname: Gong
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics*; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000235:genetics; D004847:Epithelial Cells / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D061346:Laser Capture Microdissection; D008046:Lip / Q000166:cytology* / Q000196:embryology; D008445:Maxillofacial Development / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D010159:Palate / Q000166:cytology* / Q000196:embryology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics; D053781:Transforming Growth Factor beta2 / Q000235:genetics*
chemical_list: C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D018125:Receptors, Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D017346:Protein Serine-Threonine Kinases; C498211:BMPR1A protein, human; D052005:Bone Morphogenetic Protein Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human; C000625842:Tgfbr1 protein, mouse