Cell cycle of tsAF8 after heating

Thermotolerance in tsAF8 cells develops during incubation at 34°C after heating at 4SoC, while it is suppressed by the following incubation at a non-permissive temperature of 39.7"C after the same heating. The incubation temperature after heating may affect the cell cycle and consequently thermotolerance. In the present study, a relationship between the thermotolerance and the cell cycle of tsAF8 was investigated. The cell cycle fractions and DNA synthesis were measured by flow cytometry using double staining with propidium iodide and bromodeoxyuridine. When the tsAF8 cells were heated at 4SoC for 20 min, and thereafter incubated at 34°C, bromodeoxyuridine uptake in the S phase cells (DNA synthesis) was recovered to 6S.1% 6 h after the heating, and the cells showed gradual accumulation in the G/M phase. When the cells were incubated at 39.7"C after heating at 4SoC for 20 min, then showed inhibition of thermotolerance development, the DNA synthesis was recovered to IS.1% temporarily 6 h after the heating, but it became 0% after 12 h, and the cells did not remarkably accumulate in any phases of the cell cycle. This inhibition of DNA synthesis at 39.7"C was considered to be the result of cell survival decreasing by a step-down heating. However, the relationship between the thermotolerance and the cell cycle was not found out in tsAF8 cells, because the cells did not accumulate in any phases of the cell cycle under the inhibitory condition of thermotolerance.