FRAPを用いた細胞内における化学情報、力学情報の同時計測と動態解析

抄録

Intercellular proteins are continuously exchanged by chemical reactions called turnover. Fluorescence recovery after photobleaching (FRAP) is a technique to evaluate the intercellular molecular dynamics, for exsample, turnover rate and mobile fraction in living cells. The knowledge on the molecular dynamics in entire intracellular regions can help us understand the complicated phenomenon in living cells. However, it is difficult to evaluate it because of the intrinsic limitation of FRAP experiment. Here, we applied spatial statistics to FRAP experiments to estimate the dynamics in entire intracellular regions even with limited numbers of data. Kriging is a method of interpolation of spatial statistics, and it provides a liner unbiased prediction at unsampled locations. Before interpolation using kriging, we check whether there is spatial autocorrelation in the region of interest with Molan’s I. If there is a certain level of spatial aotocorrelation, we can interpalate using kryging. We thus conducted FRAP experiments, checked the spatial autocorrelation, and interpolated with three A7r5 cells transfected with EGFP-Tublin. We found a spatial autocorrelation in the turnover rate and mobilefraction in the entire regions of some cells. Thus, our results suggest that spatial statistics is useful to know molecular dynamics in the entire intracellular region.