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: 9888191
• 负责人: Brenda L Bloodgood
• 金额: $ 33.32万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9888191
• 关键词: 5' Untranslated Regions; Action Potentials; Acute; Address; Anatomy; Animals; Apical; Behavior Control; Binding; Biology; Brain Diseases; Calcium Channel; Cell Nucleus; Cell physiology; Cells; ChIP-seq; Complex; Custom; Cyclic AMP; Data; Dendrites; Electrophysiology (science); Epilepsy; Equilibrium; Excitatory Postsynaptic Potentials; Frequencies; Gene Expression; Gene Expression Regulation; Gene-Modified; Genes; Genetic; Genetic Transcription; Genome; Genomics; Glutamates; Goals; Growth Factor; Hippocampus (Brain); Immediate-Early Genes; Inhibitory Synapse; Lead; Life; Location; Mechanics; Membrane; Mental disorders; Messenger RNA; Methods; Molecular; Monitor; Mus; N-Methyl-D-Aspartate Receptors; NPAS4 gene; Neurodevelopmental Disorder; Neurons; Optics; Output; Pattern; Phenotype; Population; Preparation; Proteins; Recording of previous events; Regulation; Schizophrenia; Signal Transduction; Slice; Stimulus; Synapses; Synaptic Potentials; Synaptic plasticity; Techniques; Testing; Training; Transcriptional Regulation; Translating; Translations; Work; autism spectrum disorder; experimental study; genome-wide; hippocampal pyramidal neuron; in vivo; insight; nervous system disorder; neuron component; neuronal cell body; neurotrophic factor; prevent; programs; recruit; response; spatiotemporal; stem; synaptic function; tool; transcription factor

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.

在神经元中，膜去极化导致即时早期基因转录因子（ieg - tf）的表达，包括NPAS4，它调节与可塑性相关的基因表达程序。在体内，eeg - tf被广泛用作识别任务相关神经元的工具，但目前尚不清楚这些蛋白质是否被诱导以响应神经元的动作电位（AP）输出或突触输入（EPSPs）的变化。至于APs和epsp是否会导致不同的基因调控模式和细胞表型，我们所知的就更少了。这种信息“传递函数”是神经元如何监测和调节自身活动的重要组成部分。NPAS4是一种调节兴奋-抑制（E-I）平衡的eeg - tf，在特定情况下，研究这种传递函数将为了解E-I平衡失调导致的神经发育和精神疾病的机制提供有价值的见解。我们已经从小鼠身上开发了一种急性海马切片制备方法，使我们能够在完整回路的背景下，从定义的输入群体中独立地唤起ap或epsp。我们建议研究NPAS4表达的活性要求，以及每种类型的NPAS4的不同基因组和突触调控