Genomic Bases of Behavioral Learning: Single Cell Approaches

行为学习的基因组基础：单细胞方法

• 批准号: 8290561
• 负责人: ROBERT D HAWKINS
• 金额: $ 41.61万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8290561
• 关键词: Affect; Afferent Neurons; Alzheimer' s Disease; Aplysia; Attention; Behavior; Behavioral; Biological Models; Brain; Cell Communication; Cells; ChIP-seq; Cyclic AMP; Cyclic GMP; Development; Disease; Drug Addiction; Enhancers; Event; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genome; Genomics; Heart; In Vitro; Individual; Interneurons; Learning; Life; Link; Logic; Maps; Memory; Memory Loss; Molecular; Molecular Target; Monitor; Motor Neurons; Neurons; Neurosciences; Output; Pathway interactions; Phenotype; Physiological; Preparation; RNA; Reflex action; Regulator Genes; Regulatory Pathway; Resolution; Role; Schizophrenia; Second Messenger Systems; Serotonin; Signal Pathway; Signal Transduction; Signaling Molecule; Synapses; Synaptic plasticity; Technology; Testing; Time; Transcriptional Regulation; Withdrawal; age related; base; chromatin remodeling; classical conditioning; conditioning; experience; genome-wide; human disease; in vivo; memory process; molecular site; neural circuit; neurodevelopment; operation; promoter; relating to nervous system; second messenger; transcription factor

DESCRIPTION (provided by applicant): Experience-dependent synaptic plasticity and underlying gene regulation are crucial for normal brain development and learning, and are disrupted in a broad array of disorders of development and learning such as schizophrenia, Alzheimer's disease, drug addiction and age related memory loss. To date, most studies of the cellular and molecular mechanisms of synaptic plasticity and gene regulation have taken a highly reductionist approach in very simplified preparations in vitro. However, practically nothing is known about genomic mechanisms of memory that occur during actual behavioral learning in the intact brain. In particular, the reductionist approach makes it difficult or impossible to study the integration of different inputs and pathways that is critical for many aspects of learning, and is a hallmark of neurodevelopment and associative learning. For these reasons, our long-term objectives are to characterize genome-wide mechanisms of long-term plasticity at the level of single identified neurons during behavioral learning. For this challenging task we will use the well-defined model system of the Aplysia withdrawal reflex, with a nearly complete mapping of a simple memory-forming circuit in a simplified behavioral preparation. We will record the activity of key individually identified neurons in that circuit and the synaptic connections between them during both a nonassociative form of learning (sensitization) and an associative form of learning (classical conditioning). And, for the first time, we will monitor the operation of the entire genome within specific individual neurons as they learn and remember. As a result, we will link neural activity to gene expression, plasticity, and behavior. We will also identify neuron type specific cellular signaling mechanisms and gene regulatory pathways during sensitization and conditioning, and test their roles in long-term plasticity. Based on our previous results, we hypothesize that three signaling pathways (5-HT, NO, and activity) act synergistically to produce more specific and longer-lasting memory traces during conditioning than during sensitization. Furthermore, 5-HT and NO can each change expression at least a thousand genes (some of which overlap) and induce large-scale chromatin remodeling. These findings have raised a fundamental question: how are these different inputs integrated at the level of genome-wide gene regulation in individual neurons in the circuit for conditioning? We will identify critical molecular targets (including promoters, enhancers and relevant transcription factors) leading to such integration, and examine their roles as decision points in the formation of long-lasting memories. These studies will significantly advance our understanding of synaptic and genomic mechanisms that contribute to circuit formation, learning, and memory, and their possible dysfunction in diseases that affect neurodevelopment and memory.

描述（由申请人提供）：经验依赖性的突触可塑性和潜在的基因调节对于正常的脑发育和学习至关重要，并且在各种各样的发展和学习障碍中受到破坏，例如精神分裂症，阿尔茨海默氏病，阿尔茨海默氏病，吸毒和年龄相关的记忆损失。迄今为止，大多数对突触可塑性和基因调节的细胞和分子机制的研究在体外非常简化的制剂中采用了高度还原的方法。但是，几乎没有关于完整大脑实际行为学习过程中发生的记忆的基因组机制的了解。特别是，还原主义的方法使研究不同输入和途径的整合对于学习的许多方面至关重要，并且是神经发展和联想学习的标志，这是难以或不可能的。 由于这些原因，我们的长期目标是表征行为学习过程中单个鉴定神经元水平上长期可塑性的全基因组机制。对于这项具有挑战性的任务，我们将使用定义明确的Aplysia撤回反射模型系统，并在简化的行为准备中几乎完整地映射了简单的内存形成电路。我们将记录关键在该电路中单独鉴定的神经元的活动，以及在非缔合形式的学习形式（敏化）和学习的联想形式（经典条件）中它们之间的突触连接。而且，我们将第一次监测特定单个神经元学习和记住的整个基因组的操作。结果，我们将神经活动与基因表达，可塑性和行为联系起来。我们还将在致敏和调节过程中确定神经元类型的特定细胞信号传导机制和基因调节途径，并测试其在长期可塑性中的作用。 基于我们以前的结果，我们假设三个信号通路（5-HT，NO和活动）协同作用，在调节过程中比在敏化过程中产生更特异性，更持久的记忆痕迹。此外，每个5-HT和NO都可以改变至少一千个基因（其中一些重叠）并诱导大规模的染色质重塑。这些发现提出了一个基本问题：这些不同的输入如何在电路中各个神经元中的全基因组调节水平集成以进行调节？我们将确定导致这种整合的关键分子靶标（包括启动子，增强子和相关转录因子），并检查其作为持续记忆形成的决策点的作用。这些研究将大大提高我们对有助于电路形成，学习和记忆的突触和基因组机制的理解，以及它们在影响神经发育和记忆的疾病中可能的功能障碍。