Quantitative determination of gap junction intercellular communication by scrape loading and image analysis
Quantitative determination of gap junction intercellular communication by scrape loading and image analysis
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DOI：
10.3109/15419060009109019
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发表时间：
2000-01-01
期刊：
影响因子：
--
通讯作者：
Rivedal, E
中科院分区：
文献类型：
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作者：
Opsahl, H;Rivedal, E
缝隙连接细胞间通讯(GJIC)由低分子分子的细胞间交换组成。化学诱导的这种通讯的改变被认为是导致细胞异常生长和肿瘤促进的结果。已经发展了几种体外试验来确定化学物质对培养细胞缝隙连接通讯的影响。刮取装载染料转移技术是基于研究荧光染料荧光黄在细胞中的转移，其中通过在细胞单层中的切割来装载该染料。这项技术快速且相对简单，但由于缺乏适当的量化染料扩散的方法，仅用于定性地展示交流。本研究表明，荧光染料标记细胞的数字荧光图像分析可用于GJIC的定量测定。我们通过染料在细胞单层中的扩散距离以及染料偶联细胞的面积来分析图像。该方法与微量注射荧光黄的结果一致，为GJIC的定量测定提供了一种简便的方法。
Gap junction intercellular communication (GJIC) consists of intercellular exchange of low molecular weight molecules. Chemically induced alterations of this communication have been suggested to result in abnormal cell growth and tumour promotion. Several in vitro assays have been developed to determine the effect of chemicals on gap junction communication in cultured cells. The scrape loading dye transfer technique is based on studying the transfer of the fluorescent dye Lucifer Yellow in cells where the dye is loaded through a cut in the cell monolayer. This technique is rapid and relatively uncomplicated, but has only been used to qualitatively demonstrate communication, due to lack of an appropriate method for quantification of the dye spreading. We show here that analysis of digital fluorescence images of cells scrape loaded with Lucifer Yellow can be used for quantitative determination of GJIC. We have analysed the images both by means of distance of diffusion of the dye in the cell monolayer, as well as by area of dye-coupled cells. The results are consistent with that obtained using microinjection of Lucifer Yellow and the method offers a simple way for quantitative determination of GJIC.