[证实了用fura-2滴定法测定MDCK细胞中镉内流的方法。]荧光视频显微镜研究]。

M Grouselle, A Boudou, B Oreja-Erroz
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引用次数: 0

摘要

采用一种基于fura-2滴定的原始方法分析了镉在哺乳动物肾细胞(MDCK)中的内流速率。该方法依赖于荧光团对金属的高亲和力。由此可见,fura-2的激发谱位移与镉的内流速率呈线性相关。荧光数字成像显微镜允许在单细胞和细胞内有机物水平的研究。结果表明,镉的吸收似乎与载体有关。金属通量与电位无关,其温度效应的Q10为2.3 +/- 0.2。维拉帕米没有效果;然而,镉的转运受到外源钙的抑制。在20℃下,镉吸收的表观解离常数估计为4.5 10(-5)M,另外一个被动跨膜扩散过程也得到了证明。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Method of measurement of cadmium influx by fura-2 titration in MDCK cell evidenced. Fluorescence video-microscopy study].

Cadmium influx rate in mammal kidney cells (MDCK) is analyzed using an original method based on fura-2 titration. The method relies on the high affinity of the fluorophore for the metal. It follows that the excitation spectrum shift of fura-2 can be linearly correlated to the influx rate of cadmium. Fluorescence digital imaging microscopy allows the study at single cell and intra-cellular organite levels. Results show that the cadmium uptake seems to be carrier dependent. Metal fluxes are potential independent, with a temperature effect caracterized by a Q10 of 2.3 +/- 0.2. No effect of verapamil is noticed; however, cadmium transport is inhibited by external calcium. Apparent dissociation constant for the cadmium uptake is estimated at 4.5 10(-5) M at 20 degrees C. An additional passive transmembrane diffusion process is also evidenced.

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