Modulation of Oligodendrocyte Development by Voltage-Operated Calcium Channels

电压驱动钙通道对少突胶质细胞发育的调节

• 批准号: 10539341
• 负责人: Pablo Martin Paez
• 金额: $ 39.88万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10539341
• 关键词: Adult; Affect; Animals; Brain; Calcium Channel; Cell Communication; Cell Fractionation; Cell Maturation; Cell membrane; Cells; Central Nervous System; Communication; Complex; Cre lox recombination system; Data; Degenerative Disorder; Dendritic Spines; Designer Drugs; Development; Electrophysiology (science); Foundations; Future; G-Protein-Coupled Receptors; Gene Expression; Genes; Genetic Recombination; Goals; Human; Image; Immunohistochemistry; In Vitro; Investigation; Ion Channel; Ions; Long-Term Potentiation; Mediating; Membrane Potentials; Molecular; Morphology; Mus; Muscarinic Acetylcholine Receptor; Mutation; Myelin; Nerve Tissue; Neurons; Oligodendroglia; Pathway interactions; Point Mutation; Potassium Channel; Process; Proliferating; Proteins; Proteomics; Research; Rest; Role; Signal Transduction; Stem Cell Development; Structure; Synapses; System; Techniques; Technology; Testing; Time; Timothy syndrome; Work; cell type; designer receptors exclusively activated by designer drugs; electrical property; experimental study; gain of function; genetic technology; improved; in vivo; in vivo imaging; insight; inward rectifier potassium channel; knock-down; migration; mouse development; mouse model; myelin degeneration; myelination; nerve stem cell; oligodendrocyte progenitor; postnatal; postnatal development; postsynaptic; remyelination; repaired; stem cells; transcription factor; transcriptome sequencing; voltage; white matter damage

PROJECT SUMMARY/ABSTRACT Accumulating evidence implicate Cav1.2 voltage-gated Ca++ channels in regulating dendritic spine morphology and thereby postsynaptic stability in neurons. Cav1.2 channels form signaling complexes in postsynaptic dendrites and dendritic spines, and functionally interact with several synaptic proteins. We have recently established that the activity of these Ca++ channels is crucial for the adequate migration, proliferation and maturation of oligodendrocyte progenitor cells (OPCs). Furthermore, our preliminary data suggest that Cav1.2 activity is associated with the expression of synaptic proteins in OPCs and is essential for the normal interaction of OPCs with neurons. Thus, we hypothesize that Cav1.2 channels that function in synaptic communication between neurons also mediate synaptic signaling between neurons and OPCs. In this research plan, we will employ imaging and electrophysiological techniques to study how Cav1.2 channels modulate the formation of synaptic connections between OPCs and neurons. We will determine whether increase Cav1.2 activity is sufficient to stimulate OPC synaptic connectivity and we will study how the activity of these channels modulates the expression of genes associated with OPC development. Three specific Aims are proposed: in the first Aim, we will employ the pseudopod subcellular fractionation system in combination with proteomics and RNA-Seq to investigate how the activity of Cav1.2 channels affect the expression of genes associated with OPC synaptic connections. Then, we will examine by electrophysiology and Ca++ imaging the synaptic connectivity of cortical OPCs in which Cav1.2 channels and specific synaptic proteins will be knock-down. In the second Aim, we will use a mouse model in which overactive Cav1.2 channels will be expressed in OPC at different postnatal time-points. The development and synaptic connectivity of these OPCs will be studied by a combination of techniques such as electrophysiology and RNA-Seq. Finally, we propose to use chemo-genetic technologies to influence the electrical properties of OPCs and thus OPC synaptic communications during brain development. Via Cre-mediated recombination we will express two G-protein-coupled receptors in OPCs, hM3Dq and hM4Di. We will evaluate how plasma membrane hyperexcitability (hM3Dq) and hyperpolarization (hM4Di) modify the establishment of synapses between OPCs and neurons and how these electrical changes affect the development of OPCs in the postnatal as well as in the adult brain. Decoding how OPCs can integrate and process synaptic input is of fundamental importance for understanding brain development and for improving remyelination of damaged white matter. We hypothesize that Cav1.2 channels are central components of OPC- neuronal synapses and are the principal ion channels mediating activity-dependent myelination.

项目摘要/摘要 积累的证据牵涉到CAV1.2电压门控Ca ++通道在调节树突状脊柱中 形态学，从而在神经元中突触后稳定性。 CAV1.2通道形成信号复合物 突触后树突和树突状刺，并在功能上与几种突触蛋白相互作用。我们有 最近确定，这些CA ++通道的活性对于充分迁移，增殖至关重要 少突胶质细胞祖细胞（OPC）的成熟。此外，我们的初步数据表明 CAV1.2活性与OPC中突触蛋白的表达有关，对于正常 OPC与神经元的相互作用。因此，我们假设Cav1.2在突触中起作用的通道 神经元之间的通信还介导神经元和OPC之间的突触信号传导。在这项研究中 计划，我们将采用成像和电生理技术来研究CAV1.2通道如何调节 OPC和神经元之间突触连接的形成。我们将确定是否增加Cav1.2 活性足以刺激OPC突触连通性，我们将研究这些通道的活性 调节与OPC发育相关的基因的表达。提出了三个具体目标： 第一个目的，我们将采用伪脚下亚细胞分级系统，并结合蛋白质组学和 RNA-Seq研究CAV1.2通道的活性如何影响与OPC相关的基因的表达 突触连接。然后，我们将通过电生理学和CA ++成像检查的突触连通性 CAV1.2通道和特定突触蛋白的皮质OPC将被敲除。在第二个目标中 我们将使用鼠标模型，其中活跃的CAV1.2通道将在OPC中在不同的后产后表达 时间点。这些OPC的发展和突触连接将通过结合 电生理学和RNA-seq等技术。最后，我们建议使用化学遗传技术 影响OPC的电性能，从而影响大脑发育过程中OPC突触通信。 通过CRE介导的重组，我们将在OPC，HM3DQ和HM4DI中表达两个G蛋白偶联受体。 我们将评估质膜过度兴奋性（HM3DQ）和超极化（HM4DI）如何修改 OPC和神经元之间建立突触，以及这些电动变化如何影响 OPC在产后和成年大脑中的发展。解码OPC如何集成和 过程突触输入对于理解大脑发育和改善至关重要 对损坏的白质的再髓。我们假设CAV1.2通道是OPC的中心组成部分 神经元突触，是介导活性依赖性髓鞘的主要离子通道。