Molecular and cellular mechanisms underlying activity dependent gene regulation in neurons

神经元活动依赖性基因调控的分子和细胞机制

• 批准号: 10354848
• 负责人: Brenda L Bloodgood
• 金额: $ 7.65万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10354848
• 关键词: Action Potentials; Acute; Biology; Brain Diseases; Cell physiology; Electrophysiology (science); Equilibrium; Excitatory Postsynaptic Potentials; Gene Expression; Gene Expression Regulation; Gene-Modified; Genomics; Hippocampus (Brain); Immediate-Early Genes; Lead; Mechanics; Membrane; Mental disorders; Methods; Molecular; Monitor; Mus; NPAS4 gene; Neurodevelopmental Disorder; Neurons; Optics; Output; Pattern; Phenotype; Population; Preparation; Proteins; Regulation; Schizophrenia; Slice; Stimulus; Synapses; Synaptic Potentials; Techniques; autism spectrum disorder; in vivo; insight; neuron component; programs; response; spatiotemporal; stem; synaptic function; tool; transcription factor

Project Abstract/Summary In neurons, membrane depolarization leads to the expression of immediate early gene transcription factors (IEG-TFs), including NPAS4, that regulate programs of gene expression associated with plasticity. IEG-TFs are widely used as tools to identify task-relevant neurons in vivo, yet it is unclear if these proteins are induced in response to changes in the action potential (AP) output or synaptic inputs (EPSPs) to the neuron. Even less is known about whether APs and EPSPs can lead to distinct patterns of gene regulation and cellular phenotypes. This information “transfer function” is an essential component of how neurons monitor and regulate their own activity. In the specific case of NPAS4, an IEG-TF that regulates excitatory-inhibitory (E-I) balance, studying this transfer function will provide valuable insight into the mechanisms underlying neurodevelopmental and psychiatric disorders that stem from dysregulation of E-I balance. We have developed an acute hippocampal slice preparation from the mouse that allows us to independently evoke APs or EPSPs, from defined populations of inputs, within the context of an intact circuit. We propose investigating the activity requirements for NPAS4 expression and the divergent genomic and synaptic regulation that follows from each type of stimulus. We have used this approach to demonstrate that APs and EPSPs lead to NPAS4 expression with distinct spatio-temporal profiles and have extensive preliminary results characterizing the unexpected underlying mechanisms. Using the methods developed for this proposal, in combination with electrophysiology, optical, and sequencing techniques, we are poised to determine how APs and EPSPs differentially impact activity-dependent gene regulation and synapse function. This proposal is a significant departure from how IEG-TFs are typically studied. The execution of these aims will yield important new insights into the mechanics of activity-dependent gene regulation in neurons and how this biology is disrupted in disorders of the brain such as Autism Spectrum Disorders and schizophrenia.

项目摘要/摘要 在神经元中，膜去极化导致立即早期基因的表达， 转录因子（IEG-TF），包括NPAS 4，调节基因表达程序 与可塑性有关。IEG-TF被广泛用作识别任务相关神经元的工具， 然而，目前还不清楚这些蛋白质是否是响应于动作电位的变化而被诱导的 (AP)输出或突触输入（EPSP）到神经元。关于AP是否 EPSP可以导致不同的基因调控模式和细胞表型。这 信息“传递函数”是神经元如何监测和调节的重要组成部分 自己的活动。在NPAS 4的特定情况下，调节兴奋-抑制性免疫应答的IEG-TF是一种免疫调节剂。 (E-I)平衡，研究这种传递函数将提供有价值的洞察机制 源于E-I失调的潜在神经发育和精神障碍 平衡我们已经开发了一种小鼠急性海马切片制备， 允许我们独立地唤起AP或EPSP，从定义的输入人群中， 一个完整的电路。我们建议调查NPAS 4的活性要求 表达和不同的基因组和突触调控，从每种类型的 刺激。我们已经使用这种方法来证明AP和EPSP导致NPAS 4 表达具有不同的时空分布，并有广泛的初步结果 描述了意想不到的潜在机制。使用为此开发的方法， 建议，结合电生理学，光学，和测序技术，我们是 准备确定AP和EPSP如何不同地影响活性依赖基因 调节和突触功能。这一提议与国际环境小组工作队的做法有很大不同， 典型研究这些目标的实现将产生重要的新见解， 神经元中活性依赖性基因调控的机制，以及这种生物学在神经元中是如何被破坏的。 自闭症谱系障碍和精神分裂症等脑部疾病。