| Cadherins are calcium-dependent, single-pass transmembrane glycoproteins responsible for cell adhesion. Classical cadherins, E-, P-, and N-cadherins, have five extracellular repeats that bind calcium ions in order to keep the extracellular portion of the molecule rigid. The cytoplasmic region of the cadherin molecule is indirectly linked to the actin cytoskeleton via plaque proteins called catenins. The cadherin molecule on one cell to binds a cadherin molecule on an adjacent cell, thus linking the actin cytoskeletons of the cells and creating an adherens junction.; Changes in gene expression and morphology allow keratinocytes, the primary cells of the epidermis, to migrate through the epidermal layers. Keratinocytes express E- and P-cadherins. To identify cadherin dependent gene expression, keratinocytes were treated with blocking antibodies to E- and/or P-cadherin, or no antibodies as a control, and RNA was extracted for differential display and gene microarray analysis. Northern blots were performed to verify differentially expressed genes.; FGF-BP was verified to be differentially expressed in keratinocytes. HaCaT cells were chosen for further analysis since they express FGF-BP and are a keratinocyte cell line. Cytosolic portions of antibody treated HaCaT cells were examined for an increase in β-catenin. Beta-catenin and N-cadherin levels were found to be decreased in the anti-Ecadherin treated cells as compared to control. However, FGF-BP expression was not increased as in primary keratinocytes. FGF treatment of HaCaT cells showed a decrease in FGF-BP transcription in bFGF and aFGF treated cells. Northern analysis indicated that there may be a decrease in N-cadherin in bFGF treated cells.; Since PKC is involved in the FGF-BP transcription pathway and in E-cadherin distribution, HaCaT cells were treated with PMA and H7. Northern blot analysis showed that FGF-BP transcription may be increased in cells treated with both PMA and H7, and N cadherin appeared to be decreased in the H7 treated cells. Immunofluorescence supports the Northern analysis that N-cadherin is decreased in the H7 treated cells. These findings could suggest ways that FGF-BP and N-cadherin transcription are regulated. |
