high-throughput screening method for measuring the activity of gap junctional intercellular communication

摘要

The present invention relates to a high speed screening method which measures gap junction activity between receptor cells of expressing iodide sensor fluorescence protein and donor cells of expressing an iodide transporter. More specifically, iodide enters donor cells and is diffused to receptor cells through gap junctions when iodide is added after coculturing receptor cells of expressing iodide sensor YFP^QL protein and donor cells of expressing iodide transporter film protein; the moved iodide can quench fluorescence by being combined with YFP^QL protein and enables activity changes of gap junctions to be checked and measured sensitively and quickly by measuring the quenching; and the intercellular gap junction activity measuring method can be used in a drug screening method using a gap junction activity controller usefully. The gap junction activity measuring method of the present invention: has a wide range of application fields since the method can be applied to cells expressing connexin protein, which is tube protein forming gap junctions; and is effective as a high speed screening analysis by expressing Z value (Zfactor) which is equal to or more than 0.7. The intercellular gap junction activity measuring method includes: a step of producing receptor cells; a step of coculturing; a step of adding buffer solution to a culturing plate; a step of calculating a quenching ratio; and a step of determining increase of gap junction activity when the calculated quenching ratio increases.