Bischof J C, Padanilam J, Holmes W H, Ezzell R M, Lee R C, Tompkins R G, Yarmush M L, Toner M
Surgical Services, Massachusetts General Hospital, Boston, USA.
Biophys J. 1995 Jun;68(6):2608-14. doi: 10.1016/S0006-3495(95)80445-5.
A quantitative fluorescent microscopy system was developed to characterize, in real time, the effects of supraphysiological temperatures between 37 degrees and 70 degrees C on the plasma membrane of mouse 3T3 fibroblasts and isolated rat skeletal muscle cells. Membrane permeability was assessed by monitoring the leakage as a function of time of the fluorescent membrane integrity probe calcein. The kinetics of dye leakage increased with increasing temperature in both the 3T3 fibroblasts and the skeletal muscle cells. Analytical solutions derived from a two-compartment transport model showed that, for both cell types, a time-dependent permeability assumption provided a statistically better fit of the model predictions to the data than a constant permeability assumption. This finding suggests that the plasma membrane integrity is continuously being compromised while cells are subjected to supraphysiological temperatures.
开发了一种定量荧光显微镜系统,以实时表征37摄氏度至70摄氏度之间的超生理温度对小鼠3T3成纤维细胞和分离的大鼠骨骼肌细胞质膜的影响。通过监测荧光膜完整性探针钙黄绿素的泄漏随时间的变化来评估膜通透性。在3T3成纤维细胞和骨骼肌细胞中,染料泄漏的动力学均随温度升高而增加。从双室转运模型得出的分析溶液表明,对于这两种细胞类型,与恒定通透性假设相比,时间依赖性通透性假设能使模型预测与数据在统计学上更好地拟合。这一发现表明,当细胞处于超生理温度时,质膜完整性不断受到损害。