LOCAL TRANSLATION IN ASTROCYTES

星形胶质细胞中的局部翻译

基本信息

批准号：
10152678
负责人：
JOSEPH D DOUGHERTY
金额：
$ 33.36万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10152678
关键词：
3&apos Untranslated Regions Acute Address Affinity Chromatography Astrocytes Biochemical Biological Assay Brain Complement Cues Data Dendrites Dependence Disease Distal Fragile X Syndrome Generations Image In Situ Hybridization In Vitro Individual Intellectual functioning disability Learning Maintenance Measures Mediating Mediator of activation protein Memory Messenger RNA Methodology Methods Modeling Molecular Morphology Mus Neurologic Neurons Open Reading Frames Peripheral Phase Physiological Process Protein Biosynthesis Proteins Regulation Ribosomes Role Seizures Signal Transduction Slice Supporting Cell Synapses System Testing Transcript Translating Translation Process Translations Untranslated Regions Work circadian density experience extracellular imaging approach improved in vivo nervous system disorder novel postsynaptic presynaptic response transcriptome sequencing translatome

项目摘要

Dysregulation of synapses is thought to underlie a range of neurological disorders, and astrocytes are key mediators of synaptic maintenance, formation and elimination. Neurons form more synapses when co- cultured with astrocytes, and a single astrocyte may ensheath 100,000 synapses in vivo. This critical role led to the “tripartite synapse” model: the synapse is not only comprised of the presynaptic bouton and postsynaptic density but also includes a peripheral astroglial process(PAP). Like dendrites and boutons, the PAP has a specific protein composition and is remodeled morphologically in response to local activity. Extensive work has demonstrated that synaptic remodeling is regulated by local translation in neurons, particularly in the dendrite. Specific mRNAs are shuttled into dendrites where their translation can be stimulated by a variety of extracellular signals, allowing for fine-tuned regulation of local protein synthesis. Newly synthesized proteins are thought to mediate the strengthening of individual synapses that underlies learning and memory, as inhibiting translation also inhibits formation of memory. As both PAPs and dendrites are remodeled in response to activity, it is likely that both responses are essential to increasing the overall efficacy of individual synapses in response to experience. Thus a key question in the field is are astrocytes, like neurons, also able to localize specific mRNAs to their processes for translation? Here, we will address this question using biochemical approaches and functional assays to test for the enrichment and translation of specific mRNAs in the PAP. We will seek to: 1) identify the transcripts localized to astroglial ribosomes in processes, 2) determine whether features in the sequence of the transcripts themselves mediate their local translation in astrocytes, 3) determine whether molecular cues of neuronal activity are capable of stimulating local translation in astrocytes.

突触的失调被认为是一系列神经系统疾病的基础，星形胶质细胞是突触维护，形成和消除的关键介体。神经元时会形成更多的突触用星形胶质细胞培养，单个星形胶质细胞可能会在体内产生100,000个突触。这个关键角色导致了 “三方突触”模型：突触不仅累积了突触前胸部和突触后的突触密度但还包括外围星形胶质过程（PAP）。像树突和座托一样，帕普有一个特异性蛋白质组成，并在形态学上对局部活性进行重塑。广泛的工作表明，突触重塑受神经元中局部翻译的调节，特别是在树突中。特定的mRNA被穿梭成树突，可以刺激其翻译通过各种细胞外信号，允许对局部蛋白质合成的微调调节。新综合蛋白质被认为可以介导研究和记忆，因为抑制翻译也抑制了记忆的形成。随着pap和树突的去除为了响应活动，这两种响应可能对提高个体的整体效率至关重要响应经验的突触。该领域的关键问题是星形胶质细胞，例如神经元，也能够将特定的mRNA定位到其翻译过程中？在这里，我们将使用生化解决这个问题方法和功能分析以测试PAP中特定mRNA的富集和翻译。我们将寻求：1）确定在过程中本地定位于星形胶质核糖体的成绩单，2）确定是否确定是否是否成绩单序列的特征本身介导了星形胶质细胞中的局部翻译，3）确定神经元活性的分子提示是否能够刺激星形胶质细胞中的局部翻译。