Mechanisms of Glutamate Receptor Synaptic Clustering

描述（由申请人提供）：该奖项的目标是支持Dane Chetkovich博士作为一名独立的医学科学家的职业生涯。这位候选人将继续他对谷氨酸受体（GluR）突触簇的研究，这是他在旧金山弗朗西斯科加州分校大卫·布雷特博士的指导下发起的一个项目，并在西北大学自己的实验室继续进行。GluRs是负责大脑中大部分兴奋性通信的蛋白质。神经元通信效率的变化是通过改变突触处GluR的数量或性质来介导的，并且是学习和记忆的基础。此外，GluR与许多疾病的病理生理学有关，包括阿尔茨海默病。Stargazin是一种GluR结合蛋白，在癫痫和共济失调的stargazer小鼠中发生突变。在观星鼠的大脑中，GluR是正常产生的，但它不以突触为目标。Chetkovich博士先前的工作表明，Stargazin将GluRs传递到突触取决于Stargazin的几个部分，这些部分与其他蛋白质相互作用，包括一种称为nPIST（与TC 10特异性相互作用的神经元蛋白质）的蛋白质。这种结合发生在stargazin的一个区域，该区域对于GluR正确靶向突触至关重要。为了进一步探索nPIST在GluR靶向突触中的作用，Chetkovich博士将使用细胞生物学、生物化学和电生理学方法来检验以下假设：nPIST分子通过将nPIST/stargazin/GluR受体复合物递送至突触支架蛋白来陪伴GluR受体靶向突触，并且nPIST结构域的磷酸化调节这一过程。该项目的具体目的是：1）了解nPISTPDZ结构域相互作用在GluR突触簇中的作用; 2）探索nPIST磷酸化在GluR突触簇中的作用; 3）评估Stargazin家族成员与nPIST和GluR亚基在GluR突触簇中的相互作用。这些实验将增强对GluR靶向突触的基本机制的理解，并应提供对异常GluR靶向导致神经系统疾病的机制的新见解。这些研究将有希望导致发现，确定新的目标，为治疗发展，以打击致残性神经系统疾病，如阿尔茨海默氏病。除了该项目在人类疾病重要领域取得科学进步的目标外，K 02还将使Chetkovich博士有受保护和结构化的研究时间，继续他的工作，指导他的学员，并在他发展科学事业的过程中与科学同事互动。西北大学为这些研究提供了出色的环境，神经病学系为Chetkovich博士提供了资源，使其成为一名独立的医生科学家。预计Chetkovich博士的进步，由这个奖项和西北大学的动态和支持性的环境培育，将使成功的竞争R 01机制和其他资金，以进一步维持他的职业发展和科学努力。

DESCRIPTION (provided by applicant): The goal of this Award is to support Dr. Dane Chetkovich in his career as an independent physician-scientist. The candidate will continue his studies on glutamate receptor (GluR) synaptic clustering, a project he initiated under the mentorship of Dr. David Bredt at the University of California, San Francisco and has continued in his own laboratory at Northwestern University. GluRs are proteins responsible for most of the excitatory communication in the brain. Changes in neuronal communication efficiency are mediated by changing the number or properties of GluRs at synapses, and underlie aspects of learning and memory. Additionally, GluRs are implicated in the pathophysiology of many diseases, including Alzheimer's disease. Stargazin is a GluR-binding protein that is mutated in the epileptic and ataxic stargazer mice. In the stargazer mouse brain, GluR is made normally, but it does not target to synapses. Dr. Chetkovich's prior work demonstrated that stargazin delivery of GluRs to synapses is dependent on several parts of stargazin that interact with other proteins, including a protein known as nPIST (neuronal Protein interacting Specifically with TC10). This binding occurs at a region of stargazin that is critical for proper GluR targeting to the synapse. To further explore the role of nPIST in GluR targeting to the synapse, Dr. Chetkovich will use cellular biological, biochemical and electrophysiological approaches to test the hypothesis that nPIST chaperones the targeting of GluR receptors to the synapse by delivering the nPIST/stargazin/GluR receptor complex to synaptic scaffolding proteins, and that phosphorylation of the nPIST domain regulates this process. The Specific Aims of the proposed project are: 1) To understand the role of nPISTPDZ domain interactions in GluR synaptic clustering; 2) To explore the role of nPIST phosphorylation in GluR synaptic clustering; and 3) To evaluate stargazin family member interactions with nPIST and GluR subunits in GluR synaptic clustering. These experiments will enhance understanding of the basic mechanism of GluR targeting to synapses, and should provide new insight into mechanisms by which abnormal GluR targeting contributes to neurological disease. These studies will hopefully lead to discoveries that identify novel targets for the development of treatments to fight disabling neurological diseases such as Alzheimer's disease. In addition to the project's goal of scientific progress in an area important to human disease, the K02 will allow Dr. Chetkovich protected and structured research time to continue his work at the bench and mentor his trainees and interact with scientific colleagues as he develops his scientific career. Northwestern University offers an outstanding environment for these studies, and the Department of Neurology has committed the resources for Dr. Chetkovich to succeed as an independent physician-scientist. It is anticipated that Dr. Chetkovich's progress, fostered by this award and the dynamic and supportive environment of Northwestern University, will enable successful competition for R01 mechanism and other funding to further sustain his career development and scientific efforts.