Project 1 Genetic Analysis of Cortical Behavioral Plasticity

项目1 皮质行为可塑性的遗传分析

• 批准号: 8110614
• 负责人: ALCINO J. SILVA
• 金额: $ 63万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8110614
• 关键词: Affect; Anterior; Behavioral; Brain region; Breeding; Collaborations; Collection; Core Facility; Disease; Electrophysiology (science); Emotional; Event; Foxes; Gene Mutation; Genes; Genetic; Genetic Screening; Image; Imaging Techniques; Laboratories; Laser Scanning Microscopy; Learning; Lesion; Mediating; Memory; Memory impairment; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; Neocortex; Neurons; Pathway interactions; Photons; Physiological; Process; Reporting; Running; Scanning; Shock; Site; Staging; Structure; Synapses; Synaptic plasticity; System; Test Result; Testing; Training; Transgenic Organisms; Vertebral column; Viral; Visual; cell type; conditioned fear; conditioning; genetic analysis; in vivo; long term memory; mutant; neocortical; positional cloning; promoter; recombinase; somatosensory

Although in the last ten years there have been significant inroads into the molecular, cellular and systems mechanisms that mediate the early stages of contextual fear conditioning, a model of emotional memory, later stages of this process remain poorly understood. Recently our laboratory reported genetic, imaging and reversible lesion evidence that supports the idea that long-term memory for contextual conditioning depends on cortical regions, such as the anterior cingulate. Now, we propose to use a combination of genetics, transgenics, electrophysiology and 2-photon in vivo imaging to unravel the molecular and cellular mechanisms of cortical plasticity that underlie remote emotional memory. The specific aims of Project 1 are: 1- To identify genes specifically required for remote memory for contextual conditioning. In our Reverse Forward Genetic (RFG) pilot screen, out of 55 transgenics and KOs selected with a random number generator from the Jackson Laboratory collection, we found that 2 affect specifically 7-day memory (remote) for contextual conditioning, without disrupting 1-day memory (recent), general activity levels or shock reactivity. We now propose to extend this screen and test contextual fear conditioning in another 350 transgenic and KO mutants. These mutants will then be screened for somatosensory (Project 2) and visual (Project 3) plasticity deficits, as a preamble for mechanistic studies (see below) to unravel the molecular and physiological mechanisms underlying the later stages of cortical and behavioral plasticity. 2- To derive region and temporally specific mutations for genes that affect long-term memory for contextual conditioning. We propose to use the loxP/Cre recombinase system to control the cell types, brain regions and temporal expression of the mutations isolated in aim 1. The resulting mice will be tested for memory deficits and will also be studied in Projects 2 and 3. 3- To uncover cortical molecular mechanisms underlying the turnover and stability of spines in the anterior cingulate. Plasticity, including behavioral plasticity (i.e. remote memory), is thought to involve changes in neuronal structure required for consolidation and stability of stored information. We propose to use 2-photon scanning confocal in vivo imaging to examine whether mutations that affect remote memory also affect turnover and stability of spines in cortical regions required for remote memory (i.e. anterior cingulate) in trained and untrained (contextual conditioning) mutants and controls. These studies will parallel related imaging studies carried out in Projects 2 and 3. All together, the studies described here and related studies in Projects 2 and 3 will unravel fundamental molecular, cellular and structural mechanisms of how the neocortex encodes and stores information. These findings will have a key impact on how we study and treat disorders associated with emotional memory.

尽管在过去的十年里，分子、细胞和系统的研究已经取得了重大进展， 调节早期情境恐惧条件反射的机制，一种情绪记忆模型， 对这一过程的后期阶段仍然知之甚少。最近我们的实验室报告说， 以及可逆性损伤的证据，支持长期记忆对情境条件作用的观点， 取决于皮质区域比如前扣带回现在，我们建议结合使用 遗传学、转基因学、电生理学和双光子体内成像， 构成远程情感记忆基础的皮质可塑性机制。项目1的具体目标是： 1-为了识别特定的基因所需的远程记忆的上下文条件。在我们的逆转 正向遗传（RFG）试验筛选，从55个转基因和科斯中随机选择 从杰克逊实验室收集的发生器中，我们发现2特别影响7天记忆（远程） 对于情境条件反射，不破坏1天记忆（最近）、一般活动水平或休克 反应性我们现在建议扩展这个屏幕，并在另一个350人中测试情境恐惧条件反射。 转基因和KO突变体。然后，这些突变体将被筛选为体感（项目2）和视觉 （项目3）可塑性缺陷，作为机制研究的序言（见下文），以解开分子和 皮层和行为可塑性后期的生理机制。 2-推导出影响长期记忆的基因的区域和时间特异性突变， 条件反射我们建议使用loxP/Cre重组酶系统来控制细胞类型、脑区 和在AIM 1中分离的突变的时间表达。由此产生的小鼠将进行记忆测试 此外，还将在项目2和项目3中进行研究。 3-揭示前额叶棘突的更新和稳定性背后的皮质分子机制 扣带。可塑性，包括行为可塑性（即远程记忆），被认为涉及的变化， 巩固和稳定储存的信息所需的神经结构。我们建议使用双光子 扫描共聚焦体内成像，以检查影响远程记忆的突变是否也影响 皮层区域中的棘的翻转和稳定性需要远程记忆（即前扣带回）， 训练的和未训练的（情境条件作用）突变体和对照。这些研究将平行相关 在项目2和3中进行的成像研究。 总之，这里描述的研究以及项目2和项目3中的相关研究将揭示基本的 新皮层如何编码和存储信息的分子、细胞和结构机制。这些 研究结果将对我们如何研究和治疗与情绪记忆相关的疾病产生关键影响。