Mechanism of Par3-mediated regulation of dendritic spine plasticity

Par3介导的树突棘可塑性调节机制

• 批准号: 10388110
• 负责人: Mikayla Voglewede
• 金额: $ 4.68万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388110
• 关键词: Affect; Behavior; Behavioral; Binding; Binding Proteins; Biochemical; Biological Assay; Brain; Cognition; Cognitive; Complex; Conflict (Psychology); Consensus; Copy Number Polymorphism; Data; Dendritic Spines; Ensure; Exhibits; Faculty; Fellowship; Filopodia; Genes; Genetic; Glycine; Golgi Apparatus; Hippocampus (Brain); Image; Impaired cognition; Impairment; In Vitro; Intelligence; Knock-out; Laboratories; Learning; Length; Long-Term Potentiation; Mathematics; Measures; Mediating; Memory; Mentorship; Microtubule Stabilization; Microtubules; Minus End of the Microtubule; Molecular; Morphology; Mus; Mutation; Nature; Neurodevelopmental Disorder; Neurons; Neurosciences; PARD6A gene; Phosphorylation; Phosphotransferases; Play; Prosencephalon; Proteins; Regulation; Risk Factors; Role; Schizophrenia; Signaling Molecule; Signaling Protein; Single Nucleotide Polymorphism; Site; Social Behavior; Social Interaction; Stains; Structural Protein; Students; Synapses; Synaptosomes; Techniques; Testing; Tissues; Training; Up-Regulation; Variant; Vertebral column; Western Blotting; anxiety-like behavior; anxiety-related behavior; atypical protein kinase C; autism spectrum disorder; cognitive function; conditional knockout; density; excitatory neuron; experimental study; hippocampal pyramidal neuron; in vivo; in vivo two-photon imaging; innovation; insight; live cell imaging; long term memory; morris water maze; mouse model; novel; object recognition; phosphoproteomics; postnatal; postsynaptic; receptor; social deficits; synaptic function; synaptogenesis

Project Summary/Abstract Dendritic spines are small, highly polarized protrusions on excitatory neurons serving as sites of postsynaptic input. Plasticity of dendritic spines is necessary for learning, while stable dendritic spines are thought to encode long-term memories. The polarized nature of dendritic spines suggests their plasticity and stability may be mediated by polarity proteins. The Partitioning defective (Par) polarity protein 3 (Par3) regulates mature dendritic spine formation in vitro, and several single nucleotide polymorphisms (SNPs) and copy number variation (CNV) of Pard3, which encodes Par3, are associated with intelligence, schizophrenia, and autism spectrum disorder (ASD). Moreover, Par3 forms a complex with atypical protein kinase C (aPKC). A constitutively active truncated aPKC variant has been proposed as a “memory molecule,” while a full length aPKC variant is implicated in long term potentiation (LTP). Together, these data implicate Par3 in mature dendritic spine stabilization, which may play a role in cognition and social interactions. However, the in vivo mechanisms of Par3 in dendritic spine plasticity and stability remains completely unknown. To investigate the mechanisms, we developed a novel conditional knockout line that depletes Par3 in postnatal forebrain pyramidal neurons (Par3-/-). Par3-/- exhibits increased dendritic spine density and increased immature dendritic spine morphology. Phosphoproteomic analysis of Par3-/- hippocampal tissue revealed increased phosphorylation of CAMSAPs, which bind and stabilize microtubule (MT) minus-ends. Our central hypothesis is that Par3 regulates MT stability through CAMSAPs to stabilize dendritic spines, which is necessary for normal cognition and social interactions. In the absence of Par3, aPKC becomes abnormally activated leading to increased CAMSAP phosphorylation and decreased MT stability. Aim 1 investigates the hypothesis that loss of Par3 destabilizes dendritic spines leading to impaired cognitive functions and social behavior in mice. Aim 2 uses biochemical assays and live cell imaging to test the hypothesis that Par3 regulates aPKC phosphorylation of CAMSAP2 at S992, influencing MT and dendritic spine stability. Together, these experiments will elucidate the in vivo mechanisms of Par3 regulation of dendritic spine stability, cognition, and social behavior. This may provide important insight to further understand the mechanisms of neurodevelopmental disorders, such as schizophrenia and ASD. The proposed fellowship will also train the applicant in several innovative techniques in biochemical, cellular, molecular, and behavioral neuroscience. The established faculty-student mentorships will ensure proper scientific and professional training necessary to become a successful, independent neurobiologist of learning and memory.

项目总结/摘要 树突棘是兴奋性神经元上的小的高度极化的突起，作为突触后神经元突触后传导的位点。 输入.树突棘的可塑性是学习所必需的，而稳定的树突棘被认为是编码 长期记忆树突棘的极化性质表明它们的可塑性和稳定性可能是 由极性蛋白介导。Par极性蛋白3（Par 3）调节成熟树突状细胞的分化 体外棘形成，以及几种单核苷酸多态性（SNP）和拷贝数变异（CNV） 编码Par 3的Pard 3与智力、精神分裂症和自闭症谱系障碍有关 （ASD）中指定的值。此外，Par 3与非典型蛋白激酶C（aPKC）形成复合物。组成型活性截短的 aPKC变体已被提出为“记忆分子”，而全长aPKC变体涉及长时间的记忆。 术语增强（LTP）。总之，这些数据暗示Par 3在成熟的树突棘稳定中，这可能 在认知和社会互动中发挥作用。然而，Par 3在树突棘中的体内机制 塑性和稳定性仍然完全未知。为了研究其机制，我们开发了一种新的 在出生后前脑锥体神经元中耗尽Par 3的条件性敲除系（Par 3-/-）。Par 3-/-展品 增加的树突棘密度和增加的未成熟树突棘形态。磷酸化蛋白质组学 对Par 3-/-海马组织的分析揭示了CAMSAP的磷酸化增加，CAMSAP结合并稳定 微管（MT）负端。我们的中心假设是Par 3通过CAMSAP调节MT稳定性， 稳定树突棘，这是正常认知和社会交往所必需的。在没有Par 3的情况下， aPKC变得异常活化，导致CAMSAP磷酸化增加和MT减少 稳定目的1研究Par 3的缺失使树突棘不稳定导致受损的树突棘的假说。 小鼠的认知功能和社会行为。Aim 2使用生化分析和活细胞成像来测试 假设Par 3调节CAMSAP 2在S992的aPKC磷酸化，影响MT和树突棘 稳定 总之，这些实验将阐明Par 3调节树突棘的体内机制。 稳定性认知和社会行为这可能为进一步了解这些机制提供重要的见解 神经发育障碍，如精神分裂症和自闭症。拟议的研究金还将培训 申请人在生物化学，细胞，分子和行为神经科学的几个创新技术。的 既定的师生导师制将确保必要的适当科学和专业培训， 成为一个成功的，独立的学习和记忆神经生物学家。