Project background: TGFβ3 plays an important role in palatogenesis and the corresponding homozygous null (-/-) mouse exhibits a defect in the fusion of the palatal shelves, resulting in clefting of the secondary palate. Palatal shelf elevation is considered to be driven by regional accumulation and hydration of glycosoaminoglycans, principally hyaluronan (HA). In vitro, addition of TGFβ3 to adult fibroblasts stimulates HA synthesis. HA is synthesised by three enzymes, Hyaluronan synthases (Has 1,2,3).
Specific aims: To investigate the role of TGFβ3 in controlling expression of the Has enzymes, we have established cell lines where the TGFβ3 gene expression has been silenced. This allows us to monitor expression of the enzymes when TGFβ3 is knocked-down and how effectively expression can be rescued by addition of exogenous growth factors. Methodology: TGFβ3 secretion by cells will be quantified by ELISA. Cells will be exposed to exogenous growth factors and the levels of Has visualised by techniques including Western blotting and immunocytochemistry.
Subject Area: Oral health & disease