SUMMARYRelationships between the thermo-sensitive response and membrane lipid fluidity were studied using a ciliated protozoan, Paramecium multimicronucleatum. Paramecium elicits a transient membrane depolarization in response to a cooling stimulus (temperature drop). The depolarization amplitude was largest when the cooling stimulus was started from the culture temperature, whilst when cooling started at a temperature more than 5°C higher or lower than the culture temperature, only a small depolarization was induced. Therefore, the cooling-induced response was dependent on the culture temperature and its sensitivity to the cooling stimulus was highest at the culture temperature. Membrane fluidity measurements of living cells using the fluorescent dye 6-lauroyl-2-dimethylaminonaphthalene (laurdan) showed that the fluidity measured at the culture temperature was almost constant irrespective of the temperature at which the cells had been cultured and adapted, which is consistent with homeoviscous adaptation. The constant fluidity at the culture temperature quickly decreased within a few seconds of application of the cooling stimulus, and the decreased fluidity gradually readapted to a constant level at the decreased temperature within 1 h. When the constant fluidity at culture temperature was modified by the addition of procaine or benzyl alcohol, the cooling-induced depolarization was completely abolished. These results suggest the possibility that the adaptation of fluidity to a constant level and its quick decrease below the constant level activate cooling-sensitive channels to elicit the transient depolarization.
SUMMARY
The relationship between thermotolerance and membrane properties was studied by using a ciliated protozoan, Paramecium aurelia. P. aurelia is a complex of sibling species termed `syngens' whose cell morphology appear similar on microscopic examination. From the comparison of tolerance to increasing temperature among 14 syngens of P. aurelia,we selected syngens 2 and 3 as low thermotolerant examples, and syngens 8 and 10 as high thermotolerant examples. The membrane resistance of high thermotolerant syngens measured by injection of a constant inward current was greater than that of low thermotolerant syngens. Membrane fluidity measurements of living cells using the fluorescent dye,6-lauroyl-2-dimethylaminonaphtalene (laurdan) showed that the fluidity at the cultured temperature was decreased in high thermotolerant syngens compared to that of low thermotolerant syngens. However, when the temperature was increased to the killing temperature of each syngens, the fluidity was increased to almost the same level irrespective of syngen. Furthermore,analysis of fatty acids extracted from whole cells showed that the ratios of unsaturated to saturated fatty acids was smaller in high thermotolerant syngens than in low thermotolerant syngens. These results suggest that the thermotolerance of P. aurelia syngens is determined by the membrane fluidity which is related to the fatty acids composition.
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