Project background: The TGFβ-3 homozygous null (-/-) mouse has a cleft palate and treatment of palatal shelves from these knockout mice, ex vivo, with TGFβ-3 causes the palatal shelves to fuse. TGFβ-3 may signal via two pathways; SMAD pathway or PI-3 kinase/Akt pathway. Blocking of the PI-3 kinase pathway by the inhibitor LY294002 causes the medial edge epithelium to persist in the midline and the basal lamina to remain intact, thus no palatal fusion is seen. Epithelial cells of the midline disappear after fusion and one of the proposed mechanisms is epithelial-mesenchymal transition (EMT) which requires PI-3 kinase activity.
Specific aims: The project will develop an in vitro model for EMT in which cell response to exogenous TGFβ-3 will be investigated and how the activation of Akt is essential in this process. The effect of TGFβ-3 on EMT will be visualised by microscopy and protein biochemistry.
Methodology: Cell Biology, Biochemistry and Immunocytochemistry.