Potenza M N, Lerner M R
Department of Cell Biology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Pigment Cell Res. 1992 Dec;5(6):372-8. doi: 10.1111/j.1600-0749.1992.tb00565.x.
A new method for rapidly evaluating the effects of drugs on receptors that regulate intracellular cAMP in a cell line derived from Xenopus laevis melanophores has been developed. Melanophores were plated into sterile 96 well microtiter plates, and 3 days later the cells were treated with melatonin for 30 min to induce melanosome aggregation. Subsequent exposure to MSH or adrenergic agonists caused dose dependent pigment dispersion that peaked within 30 min. The cumulative pigment displacement from cells could be quantitated by using a microplate reader to measure changes in transmittance of light through the wells. The acquired data enabled detailed and reproducible dose response curves and time course analyses to be generated. In addition, the assay followed for the rapid characterization of the effects of antagonists upon the beta adrenergic receptor (beta AR). The assay has the potential to test the effects of ligands upon any receptor capable of mediating pigment translocation in the melanophore cell line.
已经开发出一种新方法，用于快速评估药物对源自非洲爪蟾黑素细胞的细胞系中调节细胞内cAMP的受体的作用。将黑素细胞接种到无菌96孔微量滴定板中，3天后用褪黑素处理细胞30分钟以诱导黑素体聚集。随后暴露于促黑素细胞激素（MSH）或肾上腺素能激动剂会引起剂量依赖性色素分散，在30分钟内达到峰值。可以使用酶标仪测量通过孔的光透射率变化来定量细胞中累积的色素位移。所获得的数据能够生成详细且可重复的剂量反应曲线和时间进程分析。此外，该测定法用于快速表征拮抗剂对β-肾上腺素能受体（βAR）的作用。该测定法有潜力测试配体对黑素细胞系中任何能够介导色素转运的受体的作用。
