Mechanical forces can alter the shape and growth of cells and tissues, and are thought to be important for organ morphogenesis. However, our ability to measure the mechanical forces imposed on a tissue is limited. Here we propose to measure the mechanical tension across a sheet of epithelial cells by generating a cadherin-based tension sensor. E-Cadherin (Ecad) is a protein responsible for cell-to-cell adhesion. In this experiment we are modifying the E-Cad gene and adding a Tension Sensor Modification (TSMod) in the cytoplasmic domain of the protein. The EcadTSMod has two fluorescent domains (CFP & TFP) that will be used to determine junctional tension. What we want to know is if membrane domains containing PCP proteins are under more or less tension than the other membrane domains that do not contain PCP proteins. After transfecting skin cells with the fluorescent domains genes, it was found that the intensity of the FRET is directly proportional to the tension between cells. During the process it was also found that in order to have an <i>in vitro</i> grown polarized tissue, the cells must be grown on an air-liquid interface.

Determining junctional tension in skin cells using Ecad-TSMod

249th National Meeting (2015)

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