Department of Life Sciences, University of Trieste, 34127 Trieste, Italy.
Chem Res Toxicol. 2012 Sep 17;25(9):1912-20. doi: 10.1021/tx300203x. Epub 2012 Aug 28.
Palytoxin (PLTX) is one of the most toxic seafood contaminants ever isolated. Reports of human food-borne poisoning ascribed to PLTX suggest skeletal muscle as a primary target site. Primary cultures of mouse skeletal muscle cells were used to study the relationship between Ca(2+) response triggered by PLTX and the development of myotoxic insult. Ca(2+) imaging experiments revealed that PLTX causes a transitory intracellular Ca(2+) response (transient phase) followed by a slower and more sustained Ca(2+) increase (long-lasting phase). The transient phase is due to Ca(2+) release from intracellular stores and entry through voltage-dependent channels and the Na(+)/Ca(2+) exchanger (reverse mode). The long-lasting phase is due to a massive and prolonged Ca(2+) influx from the extracellular compartment. Sulforhodamine B assay revealed that the long-lasting phase is the one responsible for the toxicity in skeletal muscle cells. Our data analyzed, for the first time, pathways of PLTX-induced Ca(2+) entry and their correlation with PLTX-induced toxicity in skeletal muscle cells. The cellular morphology changes induced by PLTX and the sensitivity to gadolinium suggest a role for stretch-activated channels.
海兔毒素 (PLTX) 是迄今为止分离出的毒性最强的海鲜污染物之一。人类因食用 PLTX 而中毒的报告表明,骨骼肌是主要的靶器官。本文使用原代培养的小鼠骨骼肌细胞来研究 PLTX 引发的 Ca(2+)反应与肌肉毒性损伤之间的关系。钙成像实验表明,PLTX 引起短暂的细胞内 Ca(2+)反应(瞬时相),随后是较慢且更持续的 Ca(2+)增加(持续相)。瞬时相是由于细胞内储存的 Ca(2+)释放以及通过电压依赖性通道和 Na(+)/Ca(2+)交换体(反向模式)进入细胞内。持续相是由于大量和持续的 Ca(2+)从细胞外间隙流入细胞内。磺基罗丹明 B 测定表明,持续相是导致骨骼肌细胞毒性的原因。本文首次分析了 PLTX 诱导的 Ca(2+)内流途径及其与 PLTX 诱导的骨骼肌细胞毒性之间的相关性。PLTX 诱导的细胞形态变化和对钆的敏感性表明,牵张激活通道发挥了作用。