Local protein synthesis in presynaptic plasticity

突触前可塑性中的局部蛋白质合成

• 批准号: 9396910
• 负责人: Hannah Monday
• 金额: $ 4.4万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9396910
• 关键词: Acute; Adult; Amino Acids; Axon; Binding Proteins; Biological Models; Brain; Caregivers; Cells; Child; Coupled; Cytoplasmic Granules; Cytoskeleton; Data; Development; Disease; Electrophysiology (science); FMR1; Fragile X Syndrome; Glutamates; Goals; Hippocampal Mossy Fibers; Hippocampus (Brain); Imaging Techniques; Impairment; Intellectual functioning disability; Investigation; Knock-out; Label; Laser Microscopy; Laser Scanning Microscopy; Lead; Learning; Long-Term Depression; Long-Term Potentiation; Measures; Mediating; Memory; Messenger RNA; Microscopy; Modeling; Modification; Molecular; Mutate; Mutation; Occupations; Pathology; Pathway interactions; Pharmacology; Plasticizers; Process; Property; Protein Biosynthesis; Proteins; Proteome; Proteomics; RNA-Binding Proteins; Regulation; Reporting; Research; Resolution; Ribosomal Proteins; Role; Signal Transduction; Site; Slice; Societies; Source; Structural defect; Structure; Synapses; Synaptic plasticity; Techniques; Testing; Time; Translations; United States; Work; autism spectrum disorder; dentate gyrus; disability; effective therapy; experience; experimental study; functional plasticity; granule cell; inhibitor/antagonist; insight; mossy fiber; neglect; neuronal cell body; neurotransmitter release; new therapeutic target; novel; postsynaptic; presynaptic; proteostasis; receptor; response; synaptic function; two-photon

PROJECT SUMMARY The ability of synapses to change their functional and structural properties in response to activity, called synaptic plasticity, is believed to act as the molecular basis of learning and memory and relies critically on protein synthesis. The regulation of protein synthesis is essential for normal synaptic function and is altered in synaptopathic diseases like Fragile X Syndrome (FXS) and Autism Spectrum Disorders (ASDs). Despite years of research, these diseases still have no effective treatment and place a large burden on society, since many adults with FXS/ASDs require a caregiver and cannot hold a job. Synaptic plasticity can occur as a result of postsynaptic receptor modifications or presynaptic changes in neurotransmitter release. While most studies of FXS have focused on postsynaptic plasticity mechanisms, including local protein synthesis, relatively little is known about the mechanisms underlying long-term presynaptic changes in the context of FXS. The Fragile X mental retardation protein (FMRP), mutated in FXS, is an mRNA binding protein that regulates activity-dependent local protein synthesis. FMRP is expressed presynaptically, but its role in long-term presynaptic plasticity has never been investigated. Utilizing cutting-edge imaging techniques like super- resolution microscopy, and two-photon laser scanning microscopy (2PLM) coupled with electrophysiology, this proposal will investigate presynaptic protein synthesis, and its regulation by FMRP, in activity-dependent plasticity. Fluorescent Noncanonical Amino Acid Tagging (FUNCAT) will be used to measure local presynaptic protein synthesis in the context of presynaptic plasticity. Presynaptic manipulations of protein synthesis will enable investigation of the cell-specific mechanisms that lead to enduring changes in neurotransmitter release at the presynapse. To test the role of presynaptic FMRP, a conditional knock out model of FMRP will be used to specifically delete the protein in the presynaptic cell. Two-photon microscopy and electrophysiology will enable real-time assessment of the mechanisms that govern activity-dependent presynaptic structural and functional plasticity. This proposal will uncover heretofore unknown mechanisms of long-term changes in neurotransmitter release, a previously neglected line of research, and likely generate new insights into the synaptic pathology of diseases like FXS.

项目摘要 突触改变其功能和结构特性以响应活动的能力，称为 突触可塑性被认为是学习和记忆的分子基础， 蛋白质合成蛋白质合成的调节对于正常的突触功能是必不可少的，并且在神经元内发生改变。 突触病变疾病，如脆性X综合征（FXS）和自闭症谱系障碍（ASD）。尽管多年 尽管研究进展缓慢，但这些疾病仍然没有有效的治疗方法，给社会带来了沉重的负担，因为许多 患有FXS/ASD的成年人需要照顾者，并且无法工作。突触可塑性的发生可能是 突触后受体修饰或神经递质释放的突触前变化。虽然大多数研究 FXS主要关注突触后可塑性机制，包括局部蛋白质合成，相对较少的是 我们知道FXS背景下长期突触前变化的潜在机制。的 脆性X智力低下蛋白（FMRP），在FXS中突变，是一种mRNA结合蛋白， 活性依赖性局部蛋白质合成。FMRP在突触前表达，但其在长期的作用， 突触前可塑性从未被研究过。利用先进的成像技术，如超- 分辨率显微镜和双光子激光扫描显微镜（2 PLM）与电生理学相结合， 一项提案将研究突触前蛋白质的合成，以及FMRP对其的调节， 可塑性荧光非典型氨基酸标记（FUNCAT）将用于测量局部突触前 突触前可塑性背景下的蛋白质合成。突触前对蛋白质合成的操纵 能够研究导致神经递质释放持久变化的细胞特异性机制 在突触前为了测试突触前FMRP的作用，将使用FMRP的条件敲除模型 特异性地删除突触前细胞中的蛋白质。双光子显微镜和电生理学将 能够实时评估控制活动依赖性突触前结构和 功能可塑性这一建议将揭示迄今为止未知的长期变化机制， 神经递质释放，以前被忽视的研究路线，并可能产生新的见解， FXS等疾病的突触病理学。