A New Model System for Adult Neurogenesis

成人神经发生的新模型系统

• 批准号: 10452952
• 负责人: Cassandra G Extavour
• 金额: $ 24.71万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452952
• 关键词: Address; Adult; Affect; Alzheimer' s Disease; Animal Model; Animals; Biological Models; Birth; Brain; Brain Diseases; CREB1 gene; CRISPR/Cas technology; Candidate Disease Gene; Cell Differentiation process; Cell Polarity; Cell division; Cells; Complex; Confocal Microscopy; Data; Daughter; Defect; Development; Disease; Drosophila genus; Embryo; Embryonic Development; Ensure; Exhibits; Ganglia; Gene Expression Profile; Genes; Genome; Germ; Germ Lines; Goals; Gryllidae; Homeostasis; Immunohistochemistry; In Situ Hybridization; Insecta; Laboratory culture; Lead; Learning; Light; Mammals; Memory; Modeling; Modification; Molecular; Mothers; Mushroom Bodies; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neurons; Oocytes; Patients; Play; Process; Proteins; RNA; RNA Interference; Role; Source; Testing; Therapeutic; Time; Transcriptional Regulation; Work; adult neurogenesis; base; body system; cell type; cost; experimental study; fighting; fly; gene function; genetic analysis; genetic manipulation; improved; insight; knock-down; long term memory; nerve stem cell; neuroblast; novel; novel strategies; prevent; progenitor; relating to nervous system; selective expression; self-renewal; single-cell RNA sequencing; stem cell division; stem cells; transcription factor; transcriptome

PROJECT SUMMARY In development and homeostasis, progenitor stem cells of critical organ systems often divide asymmetrically. Memory formation involves adult neurogenesis, wherein progenitor stem cells in the adult brain divide asymmetrically to produce nascent neurons. The ability to form and recall specific memories is indispensable to our survival. Patients with neurodegenerative diseases such as Alzheimer’s exhibit drastic disruptions in learning and memory. As such, elucidating the developmental and mechanistic underpinnings of memory formation is critical to identifying and preventing processes that lead to brain disease. Our long-term objective is to understand the molecular mechanisms that generate new neurons in adult brains. We have established a new model organism for tackling this problem, one that combines the neural complexity of a mammal with the ease of laboratory culture and genetic manipulation of an insect: the cricket Gryllus bimaculatus. In the cricket brain, neuroblasts are neural stem cells necessary for learning and memory. We have discovered that the conserved transcription factor CREB, which plays a central role in animal learning and memory, regulates the molecular organizer oskar, and that both genes are required for long term memory. We hypothesize that Oskar and CREB direct proper asymmetric division of neuroblasts into neurons, which allows long term memories to be established. We will test this hypothesis through pursuit of three complementary Specific Aims. In Aim 1, we will determine whether oskar regulates adult neuroblast asymmetric cell division. We will use CRISPR/Cas9 and RNAi to abrogate oskar expression and determine how this affects asymmetric cell division using in situ hybridization, immunohistochemistry, and confocal microscopy. In Aim 2, we will identify the interacting partners of Oskar within Gryllus neuroblasts. To this end, we will assess the neural expression and long-term memory function of genes known to interact with oskar in other cell types. We will also take an unbiased approach to identifying relevant oskar interactors by using single cell RNA-seq to determine the complete transcriptome of neuroblasts. In Aim 3, we will define the role of CREB in regulating neuroblast division. We will reduce CREB expression in the adult brain and examine how CREB loss affects neuroblast asymmetric cell division. The results of these experiments will shed light on the cellular mechanisms that regulate memory formation, which could lead to novel approaches to ameliorate memory-related diseases.

项目摘要 在发育和稳态中，关键器官系统的祖细胞经常分裂 不对称。记忆形成涉及成人神经发生，其中祖细胞中的干细胞 成人大脑不对称地分裂以产生新生的神经元。形成和回忆特定的能力 回忆对于我们的生存是必不可少的。神经退行性疾病的患者，例如 阿尔茨海默氏症在学习和记忆中暴露了严重的破坏。因此，阐明了发展 记忆形成的机械基础对于识别和预防过程至关重要 导致脑部疾病。我们的长期目标是了解分子机制 在成人大脑中产生新的神经元。我们已经建立了一种新的模型生物来解决这个问题 问题，将哺乳动物的神经复杂性与实验室培养的易用性和 绝缘的基因操纵：板球gryllus bimaculatus。在板球大脑中，神经细胞是 神经干细胞学习和记忆所必需的。我们发现保守的 在动物学习和记忆中起着核心作用的转录因子Creb调节 分子组织者Oskar，并且两个基因都是长期记忆所必需的。我们假设 OSKAR和CREB将神经细胞的正确不对称分裂直接为神经元，这允许长时间 要建立的术语记忆。我们将通过追求三个完整性来检验这一假设 具体目标。在AIM 1中，我们将确定Oskar是否调节成人神经细胞不对称细胞 分配。我们将使用CRISPR/CAS9和RNAi废除Oskar的表达并确定如何 使用原位杂交，免疫组织化学和共焦影响不对称细胞分裂 显微镜。在AIM 2中，我们将在Gryllus神经细胞中确定Oskar的相互作用伙伴。到 这个目的，我们将评估已知的基因的神经表达和长期记忆功能 与其他单元类型中的Oskar相互作用。我们还将采取公正的方法来识别相关 Oskar相互作用者通过使用单细胞RNA-Seq确定神经细胞的完整转录组。 在AIM 3中，我们将定义CREB在确定神经细胞分裂中的作用。我们将减少Creb 成人大脑中的表达并检查CREB损失如何影响神经细胞不对称细胞分裂。 这些实验的结果将阐明调节记忆的细胞机制 形成，这可能导致新颖的方法来改善与记忆有关的疾病。