Zinc Fluorescence Biosensing and Imaging Technology

描述（由申请人提供）：拟议工作的主要目标是开发脑中游离锌的荧光指示剂系统，其具有改进的选择性、灵敏度和细胞内和细胞外的可校准响应。目前的小分子指示剂不够灵敏、选择性不足或两者兼而有之，并且许多指示剂不能以可以原位精确校准的方式（强度比、各向异性或寿命）响应。我们的努力已经取得了一些成功，我们预计，他们将是有价值的了解锌在大脑中的生理作用在体内，以及其在缺血性中风，癫痫，阿尔茨海默病，钝性创伤的病理作用。我们的具体目标是专注于开发新的方法，并使用它们来回答锌神经生物学中的重要问题。它们包括定量成像游离锌水平的细胞核中，使用一种激发比率的方法;测量锌释放在狗缺血模型中使用一种新的光纤传感器;，测量锌水平在癫痫发作期间在大鼠癫痫模型;和定量成像锌水平的线粒体。如果成功，我们预计我们的技术将有助于理解和开发上述疾病的治疗方法。

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.