Kasai Haruo, Kishimoto Takuya, Nemoto Tomomi, Hatakeyama Hiroyasu, Liu Ting-Ting, Takahashi Noriko
Division of Biophysics, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
Adv Drug Deliv Rev. 2006 Sep 15;58(7):850-77. doi: 10.1016/j.addr.2006.07.008. Epub 2006 Aug 14.
Two-photon excitation imaging is the least invasive optical approach to study living tissues. We have established two-photon extracellular polar-tracer (TEP) imaging with which it is possible to visualize and quantify all exocytic events in the plane of focus within secretory tissues. This technology also enables estimate of the precise diameters of vesicles independently of the spatial resolution of the optical microscope, and determination of the fusion pore dynamics at nanometer resolution using TEP-imaging based quantification (TEPIQ). TEP imaging has been applied to representative secretory glands, e.g., exocrine pancreas, endocrine pancreas, adrenal medulla and a pheochromocytoma cell line (PC12), and has revealed unexpected diversity in the spatial organization of exocytosis and endocytosis crucial for the physiology and pathology of secretory tissues and neurons. TEP imaging and TEPIQ analysis are powerful tools for elucidating the molecular and cellular mechanisms of exocytosis and certain related diseases, such as diabetes mellitus, and the development of new therapeutic agents and diagnostic tools.
双光子激发成像技术是研究活组织侵入性最小的光学方法。我们建立了双光子细胞外极性示踪剂（TEP）成像技术，利用该技术可以在分泌组织的焦平面内可视化并量化所有胞吐事件。这项技术还能够独立于光学显微镜的空间分辨率来估算囊泡的精确直径，并使用基于TEP成像的量化方法（TEPIQ）在纳米分辨率下测定融合孔动力学。TEP成像已应用于代表性的分泌腺，如外分泌胰腺、内分泌胰腺、肾上腺髓质和嗜铬细胞瘤细胞系（PC12），并揭示了胞吐和胞吞作用在空间组织上意想不到的多样性，这对分泌组织和神经元的生理及病理过程至关重要。TEP成像和TEPIQ分析是阐明胞吐作用的分子和细胞机制以及某些相关疾病（如糖尿病）的有力工具，同时也有助于开发新的治疗药物和诊断工具。
