Understanding the role of Fezl in adult neural stem cell maintenance and fate

了解 Fezl 在成体神经干细胞维持和命运中的作用

• 批准号: 8230526
• 负责人: Michael Arman Berberoglu
• 金额: $ 1.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8230526
• 关键词: Adult; Alzheimer' s Disease; Animal Model; Anxiety; Apoptosis; Behavior; Biology; Birth; Brain; Brain Injuries; Bromodeoxyuridine; Cell Cycle; Cell Differentiation process; Cell Maintenance; Cell Ontogeny; Cells; Defect; Development; Disease; Embryo; Environment; Genes; Genetic Models; Growth; Hippocampus (Brain); Human; Image; Image Analysis; Laboratories; Learning; Lesion; Life; Maintenance; Mammals; Memory; Mental Depression; Molecular; Mood Disorders; Names; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Parkinson Disease; Phenotype; Physiologic pulse; Play; Population; Process; Prosencephalon; Proteins; Publishing; Research; Role; Screening procedure; Signal Transduction; Staging; Stem cells; Telencephalon; Testing; Ventricular; Work; Zebrafish; Zinc Fingers; adult neurogenesis; brain tissue; cell type; chemical genetics; dopaminergic neuron; in vivo; injured; insight; interest; multipotent cell; mutant; nerve stem cell; neurogenesis; novel; novel marker; overexpression; programs; regenerative therapy; repaired; response; response to injury; self-renewal

Neurogenesis, or the birth of new neurons, continues throughout adult life in discrete regions of the forebrain (the sub-ventricular zone and hippocampus) in mammals including humans. Adult neurogenesis contributes in a crucial way to the ability of the brain to function normally. Perturbations of this process have been implicated in affective disorders including anxiety and depression, as well as in problems related to learning and memory. Understanding the basic biology of neural stem cells and the molecular and cellular regulatory mechanisms of neurogenesis will potentially have a significant impact on repairing these disease states and promoting the maintenance of healthy brain tissues. Recently, cells with proliferative abilities and expressing neural stem cell markers have been found throughout the brain ventricular zone in zebrafish, a vertebrate genetic model organism that shares considerable similarity with mammals and is amenable to genetic/chemical screening and in vivo imaging. I am interested in investigating molecular mechanisms that regulate the specification, maintenance, and differentiation of adult neural stem cells in zebrafish, through analyzing the function of an evolutionarily conserved zinc finger-containing protein named Fezl. Fezl is expressed in forebrain embryonic progenitor cells early in development and is detected in the ventricular zone of post-embryonic and adult brains. A mutant named too few was isolated in our laboratory for its deficit in brain dopamine neurons and was later found to disrupt the fezl gene. The specific aims are as follows: 1) Test the hypothesis that Fezl demarcates the ontogeny of adult neural stem cells in the zebrafish forebrain, 2) Determine whether Fezl plays a functional role in adult neural stem cell maintenance or differentiation, and 3) Determine the contribution of Fezl-expressing adult ventricular progenitor cells to differentiated cells of the adult brain under normal conditions as well as in response to injury. Our previous studies have established that Fezl is expressed in self-renewing ventricular progenitor cells of the adult zebrafish telencephalon. To determine whether these Fezl-expressing cells are multipotent and can give rise to both neurons and glial cells, I will perform in vivo lineage tracing. To then investigate Fezl function in adult neural stem cells and neurogenesis, markers of neural stem cells and neurons will be examined in the wildtype and too few (Fezl) mutant, in conjunction with BrdU pulse-chase studies. I also propose to determine the contribution of these Fezl-expressing cells to differentiated cells of the adult telencephalon under normal conditions as well as in response to injury by performing lesion studies in conjunction with lineage tracing.

神经发生，或新神经元的诞生，在包括人类在内的哺乳动物的前脑离散区域(脑室下区和海马区)持续整个成年。成人的神经发生对大脑正常运作的能力起着至关重要的作用。这一过程的扰动被认为与包括焦虑和抑郁在内的情感障碍以及与学习和记忆有关的问题有关。了解神经干细胞的基本生物学和神经发生的分子和细胞调控机制，可能对修复这些疾病状态，促进健康脑组织的维护具有重要影响。最近，在斑马鱼的脑室区域发现了具有增殖能力并表达神经干细胞标记的细胞。斑马鱼是一种脊椎动物遗传模式生物，与哺乳动物有相当大的相似性，可以进行遗传/化学筛选和活体成像。我感兴趣的是通过分析一种进化上保守的含锌指蛋白Fezl的功能，来研究斑马鱼成年神经干细胞的规范、维持和分化的分子机制。FEZL在发育早期的前脑胚胎前体细胞中表达，在胚胎后和成年大脑的脑室区域检测到。我们实验室分离出一个名为Too Lost的突变体，原因是它缺乏脑多巴胺神经元，后来被发现扰乱了fezl基因。其具体目的如下：1)验证FEZL区分斑马鱼前脑中成体神经干细胞个体发育的假设，2)确定FEZL在成体神经干细胞维持或分化中是否发挥作用，3)确定表达FEZL的成体脑室前体细胞在正常条件下和损伤反应中对成体脑组织分化细胞的贡献。我们之前的研究已经证实，FEZL在成年斑马鱼端脑的自我更新的脑室前体细胞中表达。为了确定这些表达Fezl的细胞是否具有多能性，是否可以同时产生神经元和神经胶质细胞，我将进行体内谱系追踪。然后，为了研究Fezl在成人神经干细胞中的功能和神经发生，将结合BrdU脉冲追逐研究，在野生型和太少(Fezl)突变体中检测神经干细胞和神经元的标记。我还建议通过结合血统追踪进行损伤研究来确定这些表达Fezl的细胞在正常情况下以及在损伤反应中对成年端脑分化细胞的贡献。