Expression of Thermotolerance Following Microinjection of Glutathione Disulfide

Asynchronous Chinese hamster ovary cells were microinjected with glutathione disulfide (GSSG). Successfully injected cells were scored by coinjecting FITC-dextran with GSSG, followed by fluorescent microscopy. After microinjection, cells were incubated for 2.5 h at 37°C to permit thermotolerance development and then heated at 45°C for 40 min. Cellular heat sensitivity was quantitated by counting the number of grains per cell after labeling heated cells with tritiated amino acids and processing for autoradiography. The data show that microinjection of GSSG induced thermotolerance which increased the number of grains per cell up to 500% of controls. Cells that were exposed to similar concentrations of GSSG in culture medium without microinjection or microinjected without GSSG did not develop thermotolerance.