The Neuroprotective Gene Botch in Stroke-Induced Neurogenesis

中风诱发的神经发生中的神经保护基因失误

• 批准号: 7806700
• 负责人: Sean T Byrne
• 金额: $ 4.12万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-15 至 2012-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7806700
• 关键词: Adult; Affect; Binding; Brain; CADASIL; Cardiovascular system; Cell Death; Cells; Cerebral Ischemia; Data; Development; Disease; Embryo; Exhibits; Functional disorder; Gene Expression; Gene Mutation; Genes; Glucose; Goals; Hippocampus (Brain); Human; In Vitro; Individual; Injury; Ischemia; Ischemic Preconditioning; Knock-out; Knockout Mice; Label; Laboratories; Lead; Memory; Modeling; Molecular; Morbidity - disease rate; N-Methylaspartate; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Newborn Infant; Organ; Outcome; Oxygen; Pathway interactions; Play; Process; Recovery; Research; Retroviridae; Role; Severities; Signal Transduction; Site; Staging; Stroke; Syndrome; Techniques; Tissues; Toxic effect; adult neurogenesis; base; behavior test; cognitive recovery; deprivation; early onset; excitotoxicity; experience; gliogenesis; in vivo; injury and repair; migration; mortality; mouse model; nerve stem cell; neural precursor cell; neurogenesis; newborn neuron; notch protein; novel; overexpression; polyclonal antibody; post stroke; research study; stroke recovery; therapeutic development; therapy development; tool

DESCRIPTION (provided by applicant): Stroke is among the leading causes of mortality worldwide, yet the mechanisms underlying its pathophysiology are not fully understood. Our laboratory has found a novel gene, Botch, which confers protection from cell death in models of ischemia, and also regulates the proliferation and differentiation of neural precursor cells. Botch was identified from a functional screen for neuroprotective genes, and is upregulated by ischemic preconditioning. Futhermore, expression of the gene confers protection from oxygen glucose deprivation (OGD) and NMDA excitotoxicity in primary cortical neuron cultures. The gene is expressed during various stages of development in multiple organs including the brain and cardiovascular system. Previous data generated in our laboratory show that Botch promotes both embryonic and adult neurogenesis in vivo and neuronal differentiation in vitro. Furthermore, Botch can bind to Notchi and Notch3 and inhibit Notch signaling, a pathway known to be involved in promoting neural precursor survival and regulating cellular differentiation. Our laboratory has developed tools to elucidate the function of Botch, including a knock-out (KO) mouse, constructs for gene overexpression and knockdown, and specific monoclonal and polyclonal antibodies. We have experience in immunohistochemisty and retrovirus labeling techniques to evaluate Botch-regulated neurogensis in vivo. Furthermore, our laboratory has expertise in a mouse model of stroke that employs focal cerebral ischemia. We propose that examining Botch's role on adult neurogenesis in the context of cerebral ischemia will further our understanding of underlying cellular and molecular processes that contribute to morbidity, with the ultimate goal of developing therapies to curtail the pathological outcome of stroke. Aim 1: The effect of Botch on adult neurogenesis. Does Botch influence neuronal differentiation and integration in vivo? We hypothesize that the Botch KO mouse will display increased rates of gliogenesis and a decreased functional integration of newborn neurons. Based on preliminary data, we hypothesize that Botch over- expression will lead to increased neuronal differentiation in vivo. Additionally, we hypothesize that in behavioral testing, the Botch KO mouse will exhibit a deficit in hippocampal-dependent spatial memory. Aim 2: What is Botch's impact on neurogenesis and cell death following stroke? We hypothesize that the Botch KO mouse will experience a greater severity of injury following stroke and disregulation of post-stroke neurogenesis and recovery.

描述（由申请人提供）：卒中是全球死亡的主要原因之一，但其病理生理学机制尚未完全了解。我们的实验室已经发现了一种新的基因，Botch，它赋予缺血模型中细胞死亡的保护，并调节神经前体细胞的增殖和分化。Botch是从神经保护基因的功能筛选中鉴定的，并且通过缺血预处理上调。因此，该基因的表达赋予保护，从氧葡萄糖剥夺（OGD）和NMDA兴奋性毒性在原代皮层神经元培养。该基因在包括大脑和心血管系统在内的多个器官发育的不同阶段表达。我们实验室先前的数据显示，Botch在体内促进胚胎和成体神经发生，在体外促进神经元分化。此外，Botch可以结合Notch1和Notch3并抑制Notch信号传导，这是一种已知参与促进神经前体存活和调节细胞分化的途径。我们的实验室已经开发了工具来阐明Botch的功能，包括敲除（KO）小鼠，基因过表达和敲低的构建体，以及特异性单克隆和多克隆抗体。我们在免疫组织化学和逆转录病毒标记技术方面拥有评估体内Botch调节的神经发生的经验。此外，我们的实验室在采用局灶性脑缺血的中风小鼠模型方面具有专业知识。我们建议，研究Botch在脑缺血背景下对成人神经发生的作用，将进一步加深我们对导致发病率的潜在细胞和分子过程的理解，最终目标是开发治疗方法以减少卒中的病理结果。目的1：Botch对成体神经发生的影响。Botch是否影响体内神经元的分化和整合？我们假设Botch KO小鼠将显示胶质细胞生成速率增加和新生神经元功能整合减少。基于初步数据，我们假设Botch过表达将导致体内神经元分化增加。此外，我们假设，在行为测试中，Botch KO小鼠将表现出大脑依赖性空间记忆的缺陷。目的2：Botch对中风后神经发生和细胞死亡的影响是什么？我们假设Botch KO小鼠在中风后将经历更严重的损伤以及中风后神经发生和恢复的失调。