Project description:Genomic analysis was performed to distinguish expression changes in murine medial edge epithelial cells during both palatal adherence and epithelial-mesenchymal transformation (EMT). Assessment of changes between sample 0 hours and sample 12 hours represent palatal adherence. Changes between sample 12 hours and sample 24 hours represent EMT. During adherence, we found that genes necessary for TGF-beta/LEF-1 signal transduction upregulate, as well as those necessary for cell adhesion. During EMT, we observed the loss of epithelial markers and upregulation of many genes necessary to promote the transition to mesenchyme and cell migration.
Project description:Genomic analysis was performed to distinguish expression changes in murine medial edge epithelial cells during both palatal adherence and epithelial-mesenchymal transformation (EMT). Assessment of changes between sample 0 hours and sample 12 hours represent palatal adherence. Changes between sample 12 hours and sample 24 hours represent EMT. During adherence, we found that genes necessary for TGF-beta/LEF-1 signal transduction upregulate, as well as those necessary for cell adhesion. During EMT, we observed the loss of epithelial markers and upregulation of many genes necessary to promote the transition to mesenchyme and cell migration. Keywords = EMT Keywords = palate Keywords = LEF-1 Keywords = TGF-beta Keywords = craniofacial development Keywords: time-course
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:Palatogenesis is a complex developmental process requiring coordinated growth, elevation, and fusion of the palatal shelves. Disruption of these events results in cleft palate, one of the most common congenital craniofacial anomalies. p130Crk-associated substrate (p130Cas) is an adaptor protein involved in integrin-mediated signaling and cytoskeletal regulation; however, its roles in late embryonic development remain poorly understood because global p130Cas-deficiency leads to early embryonic lethality. In this study, we used tamoxifen-inducible conditional knockout mice to investigate the function of p130Cas during palatal development. Conditional deletion of p130Cas resulted in cleft palate characterized by impaired horizontal growth of the palatal shelves. Reduced mesenchymal cell proliferation within the palatal shelves was confirmed by Ki-67 immunostaining and EdU incorporation. Consistently, primary mouse embryonic palatal mesenchymal (MEPM) cells derived from p130Cas-deficient embryos exhibited impaired proliferation and migration in vitro. In contrast, epithelial-specific deletion of p130Cas did not result in cleft palate, indicating that p130Cas function in the palatal mesenchyme is critical for normal palatal development. Together, these findings identify p130Cas as an important regulator of palatal mesenchymal expansion during secondary palatogenesis. Impaired proliferative growth of palatal mesenchyme likely underlies the developmental basis of cleft palate in this model.