Defining native glutamate receptor diversity

摘要 谷氨酸能突触有一个异常多样化的蛋白质组，由1000多个高度保守的蛋白质组组成 蛋白质，突触蛋白的突变会导致不同的行为后果。对以下方面的了解有限 谷氨酸能突触之间的分子组织阻碍了我们对突触多样性的理解 有助于大脑功能，并导致非专利疗法，往往无效。我们的目标是推进理性 通过定义组织、功能和功能，有选择地针对大脑回路中断的治疗设计 精确突触谷氨酸受体群体的治疗潜力。本项目旨在定义 克服长期技术难题的天然谷氨酸受体复合体的组成和组织 受体生物学方面的挑战。我们研究了丘脑谷氨酸能突触的分子组织。 由于其异常多样的谷氨酸能输入和基因表达，以及它参与了各种疾病。 我们最近的工作表明谷氨酸的N-甲基-D-天冬氨酸受体(NMDAR)家族起着关键作用 丘脑的特殊信号处理任务中的受体。NMDAR是可以组装的四聚体 在各种亚基组合中具有不同的功能特性，从而具有不同的治疗潜力。 值得注意的是，我们仍然不知道天然脑组织中存在哪些亚单位组合。我们假设 NMDAR亚基优先与特定的相关大分子以特定的组合组装 导致大脑中不同亚细胞组织的复合体。拟议的工作利用创新的 包括新的生化策略、超分辨率成像和基因组编辑技术在内的方法 确定：1)天然NMDAR大分子络合物的不同组成，以及2)NMDAR如何 定位产生了突触的分子多样性。该项目的结果将指导治疗策略 正在进行的小鼠疾病模型研究以及开发亚型选择性的持续努力 NMDAR调制器。此外，这种方法还可以应用于许多神经递质受体。 因此，有可能推动许多神经疾病的治疗设计。 1

SUMMARY Glutamatergic synapses have an exceptionally diverse proteome consisting of more than 1000 highly conserved proteins, and mutations in synaptic proteins cause varied behavioral consequences. Limited knowledge of molecular organization among glutamatergic synapses hinders our understanding of how synapse diversity contributes to brain function and results in generic, often ineffective, therapies. Our goal is to advance rational therapy design that selectively targets disrupted brain circuits by defining the organization, function, and therapeutic potential of precise synaptic glutamate receptor populations. This project seeks to define the composition and organization of native glutamate receptor complexes by overcoming long-standing technical challenges in receptor biology. We study molecular organization at glutamatergic synapses in the thalamus due to its exceptionally diverse glutamatergic input and gene expression as well as its involvement in varied diseases. Our recent work suggests key roles for the N-methyl-D-aspartate receptor (NMDAR) family of glutamate receptors in the specialized signal processing duties of the thalamus. NMDARs are tetramers that can assemble in various subunit combinations with distinct functional properties, and thus distinct therapeutic potential. Remarkably, we still do not know which subunit combinations exist in native brain tissue. We hypothesize that NMDAR subunits preferentially assemble in particular combinations with specific associated macromolecular complexes that lead to distinct subcellular organization in the brain. The proposed work utilizes innovative approaches including novel biochemical strategies, super-resolution imaging, and genome editing techniques to determine: 1) the diverse composition of native NMDAR macromolecular complexes, and 2) how NMDAR localization generates molecular diversity at synapses. Outcomes of this project will guide treatment strategies in ongoing studies with mouse disease models as well as sustained efforts in developing subtype-selective NMDAR modulators. Moreover, this approach could be applied to a number of neurotransmitter receptor families, and thus has the potential to advance therapy design for many neurological diseases. 1