Astrocyte RNA degradation and cognitive function

星形胶质细胞RNA降解和认知功能

基本信息

批准号：
10585257
负责人：
Dilek Colak
金额：
$ 54.04万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-19 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10585257
关键词：
Ablation Acute Address Adult Animals Astrocytes Behavior Behavioral Behavioral Assay Bioinformatics Biological Assay Biological Process Brain Brain Diseases Categories Cell model Cells Coculture Techniques Cognition Dendrites Detection Disease Electrophysiology (science)Ensure Equilibrium Excision Extracellular Space Functional disorder Genes Genetic Glutamates Goals High-Throughput Nucleotide Sequencing Hippocampus (Brain)Image Impaired cognition Impairment Knockout Mice Knowledge Label Learning Link Literature Long-Term Potentiation Marble Measures Mediating Memory Memory impairment Mental disorders Messenger RNA Modeling Molecular Mus Mutant Strains Mice Neurocognitive Neurodevelopmental Disorder Neuroglia Neurons Ontology Pathway interactions Phenotype Play Process Protein Biosynthesis Publishing RNA RNA Degradation Regulation Regulatory Pathway Reporting Research Research Proposals Role Signal Transduction Slice Synapses Synaptic Transmission Synaptic plasticity Tamoxifen Techniques Testing Time Tissues Transcript Translating Viral Work base behavior influence behavioral impairment behavioral phenotyping cell type cognitive function cognitive performance crosslinking and immunoprecipitation sequencing design experimental study functional disability in vitro activity in vivo insight mRNA Decay mRNA Stability mRNA Transcript Degradation morris water maze mouse model multi-electrode arrays nervous system disorder neurocognitive disorder neuropsychiatric disorder new therapeutic target synaptic function transcriptome sequencing two photon microscopy two-photon

项目摘要

PROJECT SUMMARY Despite its putative link to many mental illnesses, Nonsense-Mediated mRNA Decay (NMD) represents a relatively unexplored mechanism for regulating mRNA stability in brain function. NMD functions in a tissue-, cell type- and cell-state specific manner and modulates stability of selective mRNAs to fine-tune transcript abundance. There is dearth of knowledge regarding the identity of such NMD target RNAs, particularly in cells in their normal in vivo context. A particularly large gap in the field is the cell-specific function and targets of NMD in vivo. Our recent work has established that neuronal NMD regulates GLUR1 signaling and is required for proper synaptic plasticity, cognition, and local protein synthesis in dendrites, providing fundamental insight into the neuron-specific function of NMD within the brain. To date, no study has reported a specific function for NMD nor identified NMD substrates within glial cells in the brain. Astroglial control of synaptic activity translates into regulation of cognition making astrocytes a novel therapeutic target to treat cognitive dysfunctions. However, the mechanisms through which astrocytes regulate neuronal function are not well understood. Currently, it is not known whether mRNA degradation in astrocytes contribute to the regulation of synaptic plasticity and behavior. The goal of this application is to determine the contribution of astrocytic NMD to synaptic plasticity and cognitive performance. Several predicted ‘canonical’ and ‘atypical’ NMD targets are expressed in astrocytes. Our gene ontology analysis of these predicted NMD targets identified molecular function enrichment for Ca2+ signaling. Consistent with this, we have found that disruption of NMD in astrocytes resulted in elevated Ca2+ activity in vitro. Dynamic Ca2+ transients in astrocytes have been suggested to control proper basal synaptic transmission and modulate hippocampal LTP. We have also found that conditional ablation of NMD in astrocytes impaired memory in the adult mice. Based on the published literature and our preliminary studies, we hypothesize that NMD regulates Ca2+ activity in astrocytes, and astrocytic NMD is required for proper cognitive function and behavior in the adult brain. To test this hypothesis, we propose to determine 1) whether NMD is required for different aspects of learning and memory 2) the effects of astrocytic NMD ablation on neurons (e.g., assessing neuronal network connectivity and synaptic plasticity) and 3) functional deficits of NMD-deficient astrocytes (i.e., by assessing Ca2+ activity in vivo) and in vivo NMD targets in astrocytes. We will use a combination of techniques including an inducible-genetic mouse model, behavioral assays, electrophysiology, live-animal Ca2+ imaging by two-photon microscopy, stereotaxic viral labeling, Multielectrode Array Assay, in vivo RNAseq/bioinformatics, and in vivo HITS-CLIP. The successful completion of this research will provide a coherent view of how cell-specific mRNA degradation underlies the highly regulated synaptic and cognitive function in the mammalian brain and might be valuable for providing new insights into the astrocytic mechanisms of synaptic dysfunction and neurocognitive diseases.

项目摘要尽管它与许多精神疾病的假定联系，但胡说八道介导的mRNA衰变（NMD）代表相关的意外机制，用于控制大脑功能中的mRNA稳定性。 NMD在组织中起作用细胞类型和细胞状态特定方式，并调节选择性mRNA对微调转录本的稳定性关于这种NMD靶RNA的身份的知识死亡，特别是在细胞中在正常的体内环境中。该领域中特别较大的差距是细胞特异性功能和目标 nmd in Vivo。我们最近的工作已经确定神经元NMD调节glur1信号，并且是在树突中适当的突触可塑性，认知和局部蛋白质合成所必需的，提供基本深入了解大脑内NMD的神经特异性功能。迄今为止，尚无研究报告 NMD的功能，也确定了大脑神经胶质细胞中的NMD底物。突触的星形胶质控制活动转化为认知的调节，使星形胶质细胞成为治疗认知的新型治疗靶标功能障碍。但是，星形胶质细胞调节神经元功能的机制不好理解齿。目前，尚不知道星形胶质细胞中的mRNA降解是否有助于调节突触可塑性和行为。该应用的目的是确定星形胶质细胞的贡献 NMD达到合成可塑性和认知性能。一些预测的“规范”和“非典型” NMD 靶标在星形胶质细胞中表达。我们对这些预测的NMD靶标的基因本体分析 Ca2+信号传导的分子功能富集。与此一致，我们发现NMD的破坏星形胶质细胞在体外导致Ca2+活性升高。星形胶质细胞中的动态Ca2+瞬变已经建议控制适当的基础突触传播并调节海马LTP。我们也发现星形胶质细胞中NMD的条件消融损害了成年小鼠的记忆。根据已发表的文献和我们的初步研究，我们假设NMD调节星形胶质细胞中的Ca2+活性星形胶质细胞NMD需要适当的认知功能和成人大脑的行为。测试这个假设，我们建议确定1）学习和记忆不同方面是否需要NMD 2）星形胶质细胞NMD消融对神经元的影响（例如，评估神经元网络连接和突触可塑性）和3）NMD缺陷型星形胶质细胞的功能缺陷（即，通过在体内评估Ca2+活性）和体体NMD靶标的星形胶质细胞。我们将结合包括诱导型的技术组合小鼠模型，行为测定，电生理学，通过两光子显微镜的活动画Ca2+成像，立体定位病毒标记，多电极阵列测定，体内RNASEQ/Bioinformatics和In Vivo Hits-CLIP。这项研究的成功完成将提供有关细胞特异性mRNA降解的连贯观点基础是哺乳动物大脑中高度调节的突触和认知功能，可能是有价值的提供有关突触功能障碍和神经认知疾病的星形细胞机制的新见解。