ZN2+ AND CALCIUM PERMEABLE AMPA/KAINATE CHANNELS

描述：（改编自应用程序）自从在U.C.开始。Irvine 1991年底，申请人建立了一个研究细胞的实验室， 与大脑老化相关的选择性神经变性机制。 他获得了独立的资助（R 01和私人赠款），并取得了 1996年8月晋升为副教授，终身任职。 为了解决他打算解决的复杂的机械问题， 构成他的研究的核心，这将是必要的补充， 毒性和组织学范例与荧光成像技术， 膜片钳电生理学，这两个强大的工具，研究的行为， 他目前对这方面的经验有限。 不幸的是，虽然校园里的专家可以提供技术援助， 目前繁重的教学、临床和行政职务休假 没有足够的时间来掌握这些技术。 工资支持 通过确保80%的研究时间受到保护， 对申请人的职业发展有着不可估量的帮助。 高度选择性神经退行性变的生理基础 在许多大脑老化疾病中所见的蛋白质基本上是未知的。 最近的研究 揭示了一个潜在的重要线索：虽然大多数AMPA/红藻氨酸盐 受体门控通道是Ca 2+不可渗透的，某些群体的 神经元，包括许多在阿尔茨海默氏症中优先退化的类型 疾病或缺血，表达AMPA/红藻氨酸受体门控通道， 直接Ca 2+渗透性。 目前的建议是根据我们的初步建议提出的。 研究表明，内源性突触释放的阳离子，Zn 2 + 似乎以特别快的速度渗透Ca 2 +-可渗透的AMPA通道 选择性地破坏表达这些通道的神经元。 因此，最初的实验将试图检查细胞和亚细胞 海马Zn 2+和Ca 2+通透性AMPA/红藻氨酸通道的分布 神经元 将采用组织学方法和荧光成像， 定位这些通道，全细胞膜片钳技术将被使用 以检测它们对Zn 2+和Ca 2+的渗透性。 随后的实验 将检查通过这些通道的Zn 2+渗透的后果。 两 神经毒性和生理效应将被评估，使用组织学， 荧光成像和电生理技术。 人们希望 这些实验将提供初步的线索， Zn 2+可能对疾病中的选择性神经变性具有重要意义， 以及正常的神经功能。

DESCRIPTION: (Adapted from the application) Since starting at U.C. Irvine in late 1991, the applicant has established a laboratory studying cellular mechanisms of selective neurodegeneration of relevance to the aging brain. He has obtained independent funding (R01 and private grants), and achieved promotion to the rank of Associate professor, with tenure, in August, 1996. To address the sophisticated mechanistic questions which he intends will constitute the core of his research, it will be necessary to supplement toxicity and histology paradigms with techniques of fluorescent imaging and patch clamp electrophysiology, both powerful tools for studying behaviors of living neurons with which he presently has limited experience. Unfortunately, while experts on campus can provide technical assistance, his present heavy teaching, clinical and administrative duties leave insufficient time for mastery of these techniques. The salary support provided by an ISA, by assuring 80% protected time for research, would immeasurably aid the applicant s career development. The physiologic basis for the highly selective pattern of neurodegeneration seen in many diseases of the aging brain is largely unknown. Recent studies have revealed a potentially important clue: While most AMPA/kainate receptor-gated channels are Ca2+ impermeable, certain populations of neurons, including many types that preferentially degenerate in Alzheimer's disease or ischemia, express AMPA/kainate receptor gating channels with high direct Ca2+ permeability. The present proposal follows from our preliminary studies indicating that the endogenous synaptically released cation, Zn2+ appears to permeate Ca2+-permeable AMPA channels with particular rapidity selectively damaging neurons expressing these channels. Initial experiments will thus seek to examine the cellular and subcellular distribution of Zn2+ and Ca2+ permeable AMPA/kainate channel on hippocampal neurons. Histologic approaches and fluorescent imaging will be employed to localize these channels, and whole-cell patch-clamp techniques will be used to examine their permeabilities to Zn2+ and Ca2+. Subsequent experiments will examine consequences of Zn2+ permeation through these channels. Both neurotoxic and physiologic effects will be assessed, using histological, fluorescence imaging and electrophysiological techniques. It is hoped that these experiments will provide initial clues to physiological effects of Zn2+ that may be of importance to selective neurodegeneration in disease as well as to normal neuronal functioning.