MicroRNA-132 regulation of dendritic growth

MicroRNA-132对树突生长的调节

• 批准号: 8824566
• 负责人: RICHARD H. GOODMAN
• 金额: $ 38.5万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8824566
• 关键词: Adult; Alleles; Alzheimer' s Disease; Autistic Disorder; Behavior; Behavior Control; Biological Assay; Biological Testing; Brain; Bromodeoxyuridine; CREB1 gene; Cell Culture Techniques; Cell Cycle; Cells; EGF gene; Exercise; Exposure to; Genetic; Growth; Growth Factor; Heparin Binding; Hippocampus (Brain); Individual; Investigation; Knock-out; Laboratories; Learning; Link; Measures; Mediating; Mental Retardation; Methods; MicroRNAs; Modeling; Monitor; Morphology; Mouse Strains; Mus; Neurologic; Neuronal Plasticity; Neurons; Newborn Infant; Parkinson Disease; Population; Process; Property; Regulation; Reporter; Retroviral Vector; Role; Schizophrenia; Series; Signal Transduction; Synapses; System; Testing; Transgenes; Viral; adult neurogenesis; base; conditioned fear; in vivo; in vivo Model; inhibitor/antagonist; mRNA Expression; neuronal growth; neuronal survival; newborn neuron; novel strategies; object recognition; ratiometric; relating to nervous system; research study; response; sensor; tissue culture

DESCRIPTION (provided by applicant): MicroRNAs contribute to critical brain functions and participate in multiple neurological and psychiatric conditions. Exactly how specific microRNAs relate to these processes remains unclear, however, for several reasons. First, genetic knockouts of brain-specific microRNAs have been difficult to generate. Second, predicting microRNA targets remains extraordinarily challenging. Third, methods for measuring microRNA levels in individual cells and, more importantly, linking these levels to the regulation of particuar targets are lacking. The overall objective of this application is to determine how the CREB-regulated microRNA, miR-132, directs dendritic growth and plasticity. Our specific aims are to: 1) Characterize the role of miR-132 in brain using a conditional knockout. We have already generated a mouse strain containing a floxed miR-132 allele and have shown, using a Cre-expressing retroviral vector, that newborn hippocampal neurons lacking miR-132 display a dramatic decrease in dendritic growth and branching. This was the first example of a neural-specific microRNA knockout, and we will use these mice to examine the role of miR-132 in adult neurogenesis, dendritic growth, and behaviors associated with activity-induced morphological changes, such as spatial learning and fear conditioning. 2) Determine whether heparin-binding (Hb)-EGF and the CDK inhibitor, p21, two miR-132 targets identified in Ago2-immunopurification assays, contribute to miR-132 effects on neuronal survival, morphology, and function. 3) Determine how changes in microRNA expression in individual neurons correlates with expression of specific microRNA targets. Assays of microRNA function typically average responses of large populations of cells. Using a new ratiometric microRNA sensor developed in our laboratory, we observed that individual neurons display large differences in the expression of miR-132. We will now ask whether these differences in miR-132 levels are associated with differences in target mRNA expression using, initially, cultured neurons and, subsequently, in vivo models.

描述（由申请人提供）：MicroRNA 有助于关键的大脑功能并参与多种神经和精神疾病。然而，由于多种原因，具体的 microRNA 与这些过程的具体关系仍不清楚。首先，大脑特异性 microRNA 的基因敲除很难产生。其次，预测 microRNA 靶点仍然极具挑战性。第三，缺乏测量单个细胞中 microRNA 水平的方法，更重要的是，缺乏将这些水平与特定靶点的调节联系起来的方法。本申请的总体目标是确定 CREB ​​调节的 microRNA（miR-132）如何指导树突生长和可塑性。我们的具体目标是：1) 使用条件敲除来表征 miR-132 在大脑中的作用。我们已经培育出含有 floxed miR-132 等位基因的小鼠品系，并使用表达 Cre 的逆转录病毒载体证明，缺乏 miR-132 的新生海马神经元的树突生长和分支显着减少。这是神经特异性 microRNA 敲除的第一个例子，我们将使用这些小鼠来研究 miR-132 在成人神经发生、树突生长以及与活动诱导的形态变化相关的行为（例如空间学习和恐惧调节）中的作用。 2) 确定肝素结合 (Hb)-EGF 和 CDK 抑制剂 p21（Ago2 免疫纯化测定中鉴定的两个 miR-132 靶标）是否有助于 miR-132 对神经元存活、形态和功能的影响。 3) 确定单个神经元中 microRNA 表达的变化如何与特定 microRNA 靶标的表达相关。 microRNA 功能的测定通常是大量细胞的平均反应。使用我们实验室开发的新型比例式 microRNA 传感器，我们观察到单个神经元的 miR-132 表达存在巨大差异。我们现在将首先使用培养神经元，随后使用体内模型来询问 miR-132 水平的这些差异是否与靶标 mRNA 表达的差异相关。