INVESTIGATION OF A CELL MECHANOSENSING SYSTEM BY MEASURING CYTOSKELETAL DEFORMATION AND INTRACELLULAR CALCIUM ION CONCENTRATION

摘要

This paper describes a method to investigate the relationship between cytoskeletal deformation by mechanical stimulation and its corresponding intracellular signals for identifying mechanosensors of cells. Gene transfection of green fluorescent protein to osteoblasts enabled visualization of actin in cells. When local deformation was applied to a cell by a micropipette, the distribution of cytoskeletal actin deformation in the whole cell was automatically obtained from the two images of the cell before and after deformation by using KLT method. Calcium ion signaling response to the same mechanical stimulation was measured as the spatial and temporal changes of fluorescent intensity of fluo-4 loaded to osteoblasts. As a result, we obtained the relationship between the deformation and the biochemical signals in cells.