Molecular and Cellular Mechanisms Underlying Activity Dependent Gene Regulation in Neurons

神经元活性依赖性基因调控的分子和细胞机制

基本信息

批准号：
10469796
负责人：
Brenda L Bloodgood
金额：
$ 4.55万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/10469796
关键词：
5&apos 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

项目摘要

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，膜去极化导致立即早期基因转录因子 (IEG-TF) 的表达，包括 NPAS4，它调节与可塑性相关的基因表达程序，被广泛用作识别体内任务相关神经元的工具。但尚不清楚这些蛋白质是否是响应神经元动作电位 (AP) 输出或突触输入 (EPSP) 的变化而诱导的，更不清楚 AP 和 EPSP 是否会导致不同的结果。这种信息“传递功能”是神经元如何监控和调节自身活动的重要组成部分，在 NPAS4（一种调节兴奋性抑制 (E-I) 平衡的 IEG-TF）的具体情况中。这种传递函数将为了解 E-I 平衡失调引起的神经发育和精神疾病的机制提供有价值的见解。我们开发了一种小鼠急性海马切片制剂，使我们能够独立诱发 AP 或 EPSP，我们建议在完整电路的背景下研究 NPAS4 表达的活性要求以及每种类型的不同基因组和突触调节。我们已经使用这种方法来证明 AP 和 EPSP 导致 NPAS4 表达具有不同的时空特征，并使用为此提议开发的方法结合电生理学、光学和电生理学，获得了广泛的初步结果来表征意想不到的潜在机制。通过测序技术，我们准备确定 AP 和 EPSP 如何对活性依赖性基因调控和突触功能产生不同的影响。该提议与 IEG-TF 的典型研究方式有很大不同。这些目标的执行将产生重要的结果。对神经元活动依赖性基因调控机制的新见解，以及这种生物学在自闭症谱系障碍和精神分裂症等大脑疾病中如何受到破坏。