Cryosolvent interaction with cellular actin using 3T3-LI cells as a model system.

摘要

Previous immunolocalisation studies using intact cells have identified modification of the cytoskeleton by cryoprotectants. In the present study we have used a proteomics approach to directly resolve the interactive effects of 3T3-LI cells exposed to two cryoprotectants, dimethyl sulphoxide (Me(2)SO) and 1,2-propanediol (PROH) in 5,10, 20 and 50(v/v) percent solutions, respectively. Two-dimensional protein electrophoresis and Western blot analysis of the cell extracts identified a range of immunoreactive actin fragments with varying molecular weights and isoelectric points at all cryoprotectant concentrations. The addition of either 10mM l-cysteine or reduced glutathione to the cells prior to cryprotectant exposure modified the actin fragmentation. In this preliminary report, we have provided direct evidence of actin fragmentation when exposed to cryoprotectants and have demonstrated that the use of redox agents can modify the cryprotectant action.