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，膜去极化会导致早期基因转录因子（IEG-TF）的表达，包括NPAS4，该因素调节与可塑性相关的基因表达程序。 IEG-TF被广泛用作体内识别与任务相关的神经元的工具，但是目前尚不清楚这些蛋白是否是响应动作电位（AP）输出（AP）输出或突触输入（EPSP）的变化而诱导的。关于APS和EPSP是否可以导致基因调节和细胞表型的不同模式，甚至更少知道。此信息“转移功能”是神经元如何监测和调节其自身活动的重要组成部分。在NPAS4的特定情况下，调节抑制运动的IEG-TF（E-I）平衡，研究此转移功能将为神经发育和精神疾病的基础机制提供宝贵的见解，这些机制源于E-I平衡失调。我们已经从小鼠开发了一种急性海马切片制剂，使我们能够在完整的电路中独立地从定义的输入人群中唤起APS或EPSP。我们建议研究NPAS4表达的活性要求以及每种类型的基因组和突触调节的不同刺激。我们已经使用这种方法来证明APS和EPSP导致NPAS4表达具有不同的时空特征，并且具有表征意外的潜在机制的广泛初步结果。使用为该建议开发的方法，结合电生理学，光学和测序技术，我们被毒死以确定AP和EPSP如何差异影响活性依赖性基因调节和突触功能。该建议与IEG-TF通常是研究的方式有很大的不同。这些目标的执行将产生有关神经元中活动依赖性基因调节机制的重要新见解，以及该生物学如何在大脑的疾病中破坏自闭症谱系障碍和精神分裂症。