A New Model System for Adult Neurogenesis

成人神经发生的新模型系统

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要在发育和稳态过程中，关键器官系统的祖干细胞经常分裂记忆形成涉及成体神经发生，因此涉及祖细胞。成人大脑不对称地分裂以产生新生神经元，形成和回忆特定的能力。记忆对于患有神经退行性疾病（例如，神经退行性疾病）的患者来说是不可或缺的。阿尔茨海默氏症表现出学习和记忆的严重破坏，这说明了其发育过程。记忆形成的机械基础对于识别和预防过程至关重要我们的长期目标是了解导致脑部疾病的分子机制。在成人大脑中产生新的神经元我们已经建立了一种与之相互作用的新模型生物体。问题，一个将哺乳动物的神经复杂性与实验室培养的简易性结合起来的问题昆虫的基因操作：蟋蟀 Gryllus bimaculatus 在蟋蟀大脑中，神经母细胞是。我们发现，学习和记忆所必需的神经干细胞是保守的。转录因子 CREB 在动物学习和记忆中起核心作用，调节分子组织者奥斯卡，这两个基因都是长期记忆所必需的。 Oskar 和 CREB 指导神经母细胞适当不对称分裂为神经元，这使得长期我们将通过追求三个互补来检验这个假设。具体目标在目标 1 中，我们将确定 oskar 是否调节成体神经母细胞不对称细胞。我们将使用 CRISPR/Cas9 和 RNAi 来消除 oskar 表达并确定其如何进行。使用原位杂交、免疫组织化学和共聚焦影响不对称细胞分裂在目标 2 中，我们将鉴定 Gryllus 神经母细胞中 Oskar 的相互作用伙伴。为此，我们将评估已知基因的神经表达和长期记忆功能我们还将采取公正的方法来识别相关的细胞。 oskar 相互作用子通过使用单细胞 RNA-seq 来确定神经母细胞的完整转录组。在目标 3 中，我们将定义 CREB 在调节神经母细胞分裂中的作用，我们将减少 CREB。并研究 CREB 缺失如何影响神经母细胞不对称细胞分裂。这些实验的结果将揭示调节记忆的细胞机制形成，这可能会带来改善记忆相关疾病的新方法。