Development of mouse lingual epithelium in situ Ca imaging method for evaluation of seafood tastes

我们建立了一种原位监测小鼠味觉感受器细胞Ca^<2+>反应的新方法。用酶从小鼠舌上取下舌上皮片，并加载荧光Ca^2+探针钙绿色1。通过图像增强CCD摄像机采集显微镜观察下的荧光图像序列，并在个人计算机中进行分析，重复性地获得了甜、苦和鲜三种味觉模式的剂量依赖性Ca^2+响应。目前的Ca^2+成像技术使我们能够监测长时间（超过90分钟）、实时和多细胞反应。该方法可用于海鲜等复杂味觉的评价，并为探讨鲜味感受的细胞内信号转导机制提供了一些信息。

We developed the new method for monitoring Ca^<2+> responses in the mouse taste receptor cells in situ. The lingual epithelium sheet was enzymatically removed from a mouse tongue and loaded with a fluorescent Ca^<2+> probe, Calcium Green 1. Fluorescent image sequences under microscopic observation were acquired via an image-intensified CCD camera and analyzed in a personal computer.Dose-dependent Ca^<2+> responses were obtained reproductively for three taste modalities, sweet, bitter and umami. The present Ca^<2+> imaging technique enabled us to monitor long-time (over 90 min), real-time and multi-cellular responses. This method was useful for evaluation of complicated tastes such as seafood.Some informations about intracellular signal transduction mechanisms in taste reception of umami were obtained by this method.