Zinc Fluorescence Biosensing and Imaging Technology

描述（由申请人提供）：拟开展的主要工作是开发脑内游离锌的荧光指示剂系统，提高细胞内和细胞外的选择性、灵敏度和可校准反应。目前的小分子指示剂不够敏感，选择性不够，或者两者兼而有之，而且许多指示剂的反应方式（强度比、各向异性或寿命）不能精确地在原位校准。我们的努力已经取得了一些成功，我们预计这将有助于了解锌在体内大脑中的生理作用，以及锌在缺血性中风、癫痫、阿尔茨海默病和钝性创伤中的病理作用。我们的具体目标是集中在开发新的方法，并利用它们来回答锌神经生物学的重要问题。它们包括定量成像游离锌水平在细胞核中，使用激发比率法；用新型光纤传感器测量犬缺血模型锌释放量测量大鼠癫痫模型发作期间的锌水平；以及线粒体锌含量的定量成像。如果成功，我们预计我们的技术将用于理解和开发上述条件的治疗方法。

DESCRIPTION (provided by applicant): The major thrust of the proposed work is to develop fluorescent indicator systems for free zinc in the brain, with improved selectivity, sensitivity, and calibratable response both intracellulady and extracellularly. Current small molecule indicators are insufficiently sensitive, selective, or both, and many do not respond in a fashion (intensity ratio, anisotropy, or lifetime) that can be accurately calibrated in situ. Our efforts have already met some success, and we anticipate that they will be valuable in understanding the physiological roles of zinc in the brain in vivo, as well as its pathological roles in ischemic stroke, epilepsy, Alzheimer's disease, and blunt trauma. Our specific aims are focused on developing new methods and using them to answer questions of importance in zinc neurobiology. They include quantitatively imaging free zinc levels in the cell nucleus, using an excitation ratiometric approach; measuring zinc release in a dog ischemia model using a novel fiber optic sensor;, measuring zinc levels during seizures in a rat epilepsy model; and quantitatively imaging zinc levels in mitochondria. If successful, we anticipate that our technology will be of use in understanding and developing treatments for the conditions listed above.