The growth of tissue culture TO-2 cells derived from the warm water fish Tilapia, the induction of thermotolerance, and protein synthesis profiles of these cells in response to temperature changes were examined. TO-2 cells can grow between 15 to 34 degrees, with an optimal growth temperature of 31 degrees. There is no apparent killing of the cells when the temperature is lowered to 4 degrees for up to 3 days. Survival of TO-2 cells at 43 degrees was studied after various preheat treatments: 1) acute heating at 40 degrees for 15 min followed by 31 degrees incubation, 2) chronic exposure at 37 degrees for several hr, or 3) long-term thermal adaptation at 34 degrees. The cells acquire thermotolerance from pre-exposure to 37 degrees for as short as 6 hr. Preheating at 40 degrees followed by incubation at 31 degrees also induces thermotolerance against a subsequent 43 degrees heat challenge. In addition, 34 degrees thermal adapted cells are resistant to 43 degrees heating. One- and two-dimensional gel electrophoresis of proteins after heat treatments show that three major heat shock proteins with molecular weights around 87, 70, and 27 kD are preferentially synthesized. The synthesis of two additional proteins with an isoelectric point of 6.9 and molecular weights of 60 and 44 kD are significantly enhanced in 34 degrees thermal-adapted and 37 degrees chronic heated cells, but not in cells subjected to an acute heat shock at either 40 degrees or 43 degrees. On the other hand, the 27 kD heat shock proteins are mainly present in the 43 degrees, 40 degrees, and 37 degrees heat-shocked cells, but not in the 34 degrees thermal-adapted cells.