Synaptic Organization and Modulation of Kainate Receptors: Investigating the Structure, Dynamics, and Function in the Context of Trans-Synaptic Junctions

红藻氨酸受体的突触组织和调节：研究跨突触连接的结构、动力学和功能

• 批准号: 10754481
• 负责人: Cuauhtemoc Ulises Gonzalez
• 金额: $ 3.67万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10754481
• 关键词: 3-Dimensional; Behavior; Behavioral; Binding; Biological; Calcium; Calcium Channel; Calcium Signaling; Cations; Cells; Complement; Complement 1q; Complex; Concanavalin A; Coupling; Cryoelectron Microscopy; Disease; Doctor of Philosophy; Electron Microscopy; Electrons; Electrophysiology (science); Epilepsy; Extracellular Domain; Family; Fluorescence; Fluorescence Resonance Energy Transfer; Foundations; Future; Glutamate Receptor; Glutamates; Glycine; Investigation; Ion Channel; Kainic Acid Receptors; Knowledge; Lead; Learning; Link; Manic; Measurement; Measures; Mediating; Membrane; Membrane Proteins; Mental Retardation; Methodology; Methods; Molecular Conformation; Morphology; Neurocognitive Deficit; Neurologic Deficit; Neurons; Orphan; Permeability; Phase; Physiological; Plant Lectins; Play; Postdoctoral Fellow; Postsynaptic Membrane; Process; Property; Proteins; Regulation; Research; Research Personnel; Research Project Grants; Role; Schizophrenia; Serine; Shapes; Signal Transduction; Signaling Protein; Site; Structure; Synapses; Synaptic Cleft; Synaptic Membranes; Synaptic Receptors; Techniques; Technology; Testing; Training; Work; autism spectrum disorder; biophysical properties; delta receptors; desensitization; doctoral student; experimental study; extracellular; insight; kainate; ligand gated channel; macromolecule; member; mossy fiber; neurotransmission; neurotransmitter release; novel; pharmacologic; postsynaptic; presynaptic; protein complex; protein function; protein purification; receptor; receptor function; recruit; response; scaffold; single-molecule FRET; skills; temporal measurement

Abstract Kainate receptors are members of the ionotropic glutamate receptor family and are implicated in epilepsy and in multiple neurocognitive deficits such as autism, schizophrenia, and mental retardation. These receptors can be found to express in the presynaptic membrane where they regulate neurotransmitter release and postsynaptic membrane where it is involved in excitatory signaling. These channels are calcium-permeable and, therefore, activation by glutamate lead to intracellular calcium signaling and synaptic response. However, their current activity can be modulated by protein interactions. Such interactions can lead to increase steady-state currents, causing increase calcium permeability and differential synaptic response. Recently, presynaptic Neurexin3β and extracellular scaffolding C1q-like protein have been revealed to create a junction with postsynaptic kainate receptors at mossy fibers-CA3 synapses. These interactions have been found to regulate kainate receptor recruitment at the membrane, however, their modulatory effect on kainate receptor gating properties still remains unknown. For my dissertation work, I propose to study how protein interaction at the extracellular domain of kainate receptor modulate their gating properties. As part of the F99 training, I will use electrophysiology to investigate the current modulation caused by interactions with Neurexin 3 and C1q-like protein and investigate the conformational changes using smFRET (Aim 1). This work will give us more insight into the gating mechanisms caused by the modulation of these proteins and help explain the role of these junctions in postsynaptic excitatory signaling. For my training as a post-doctoral student, I intend to continue exploring the macromolecule contacts between these junctions using electron microscopy. Therefore, for the K00 phase of this proposal I plan to train in performing CryoEM and CryoET experiments. Using these technologies will allow me to investigate the connections made between these proteins in the context of these synaptic junctions (Aim 2). The information obtained in these experiments will give us insight into the structure and organization of this junctions within the synaptic cleft and further our understanding of the role these junctions play in synapse morphology. Understanding this will allow us to correlate biological disruption of this junction to behavior deficits. In summary, the objective of this proposal is to give me the training necessary to investigate the functional and structural function of these junctions to understand their role in physiological conditions. The training proposed under this proposal alongside my previous training will give me the framework necessary to become an independent investigator.

抽象的 红藻氨酸受体是离子型谷氨酸受体家族的成员，与癫痫和 多种神经认知缺陷，如自闭症、精神分裂症和精神发育迟滞，这些受体可能与这些受体有关。 发现在突触前膜中表达，调节神经递质释放和突触后 这些通道是钙可渗透的，因此，参与兴奋性信号传导的膜。 谷氨酸的激活导致细胞内钙信号传导和突触反应。 活性可以通过蛋白质相互作用来调节，这种相互作用可以导致稳态电流增加， 导致钙渗透性增加和突触反应差异最近，突触前 Neurexin3β 和 细胞外支架 C1q 样蛋白已被发现与突触后红藻氨酸建立连接 已发现这些相互作用可调节苔藓纤维-CA3 突触上的红藻氨酸受体。 然而，它们对红藻氨酸受体门控特性的调节作用仍然存在 未知。 对于我的论文工作，我建议研究红藻氨酸胞外域的蛋白质相互作用 作为 F99 训练的一部分，我将使用电生理学来研究。 与 Neurexin 3 和 C1q 样蛋白相互作用引起的电流调节，并研究 使用 smFRET 进行构象变化（目标 1）。这项工作将使我们更深入地了解门控机制。 由这些蛋白质的调节引起，有助于解释这些连接在突触后兴奋性中的作用 发信号。 对于我作为博士后的培养，我打算继续探索之间的大分子联系 因此，对于本提案的 K00 阶段，我计划进行培训。 使用这些技术进行 CryoEM 和 CryoET 实验将使我能够研究 这些蛋白质之间在这些突触连接的背景下建立的连接（目标 2）。 在这些实验中获得的结果将使我们深入了解该连接点的结构和组织 突触间隙并进一步我们对这些连接在突触形态中所起的作用的理解。 了解这一点将使我们能够将这种连接的生物破坏与行为缺陷联系起来。 总之，该提案的目的是为我提供必要的培训，以调查功能和 这些连接的结构功能，以了解它们在生理条件下的作用。 根据这项建议以及我之前的培训将为我提供成为一名 独立调查员。