Mechanism of Par3-mediated regulation of dendritic spine plasticity

Par3介导的树突棘可塑性调节机制

基本信息

批准号：
10602456
负责人：
Mikayla Voglewede
金额：
$ 4.77万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10602456
关键词：
Affect Behavior Behavioral Binding Binding Proteins Biochemical Biological Assay Brain Catalytic Domain Cognition Cognitive Complex Consensus Copy Number Polymorphism Data Dendritic Spines Education Ensure Exhibits Faculty Fellowship Filopodia Genes Genetic Glycine Golgi Apparatus Hippocampus Image Impaired cognition Impairment In Vitro Intelligence Knock-out Knockout Mice Laboratories Learning Length Long-Term Potentiation 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 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 mathematical ability morris water maze mouse model novel object recognition phosphoproteomics postnatal postsynaptic receptor social social deficits synaptic function synaptogenesis

项目摘要

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.

项目摘要/摘要树突状棘是小的，高度极化的蛋白输入。树突状刺的可塑性对于学习是必要的，而稳定的树突状刺则被认为是编码的长期记忆。树突状刺的两极化性质表明它们的可塑性和稳定性可能是通过极性蛋白介导。分配有缺陷（PAR）极性蛋白3（PAR3）调节成熟的树突状体外形成脊柱，几种核苷酸多态性（SNP）和拷贝数变化（CNV）编码PAR3的PARD3与智力，精神分裂症和自闭症谱系障碍有关（ASD）。此外，PAR3与非典型蛋白激酶C（APKC）形成复合物。组成性主动截断 APKC变体已被提议为“内存分子”，而全长APKC变体则以长期实现期限电位（LTP）。总之，这些数据暗示着成熟的树突状脊柱稳定中的PAR3，这可能在认知和社会互动中发挥作用。但是，树突状脊柱中PAR3的体内机制可塑性和稳定性仍然是完全未知的。为了研究机制，我们开发了一种小说有条件的敲除线会在产后前脑金字塔神经元（PAR3 - / - ）中耗尽PAR3。 PAR3 - / - 展览树突状脊柱密度增加并增加了未成熟的树突状脊柱形态。磷酸化蛋白质组学 PAR3 - / - 海马组织的分析显示，凸轮saps的磷酸化增加，结合并稳定微管（MT）减去端。我们的核心假设是PAR3通过凸轮saps调节MT稳定性稳定树突状刺，这对于正常的认知和社交互动是必不可少的。在没有par3的情况下 APKC显着激活，导致CAMSAP磷酸化增加并减少MT 稳定。 AIM 1调查了以下假设：par3的损失破坏了树突状的棘突小鼠的认知功能和社会行为。 AIM 2使用生化测定和活细胞成像来测试 PAR3在S992上调节CAMSAP2的APKC磷酸化的假设，影响MT和树突状脊柱稳定。总之，这些实验将阐明树突状脊柱调节PAR3的体内机制稳定，认知和社会行为。这可能会提供重要的见解，以进一步了解机制神经发育障碍，例如精神分裂症和ASD。拟议的奖学金还将训练在生化，细胞，分子和行为神经科学中的几种创新技术中的申请人。既定的教师精神训练将确保必要的适当的科学和专业培训成为学习和记忆的成功，独立的神经生物学家。