Regulation and regeneration of hippocampal plasticity by neural progenitor cells

神经祖细胞对海马可塑性的调节和再生

• 批准号: 9146436
• 负责人: Elizabeth Diana Kirby
• 金额: $ 9.21万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9146436
• 关键词: Acute; Adult; Age; Antiepileptic Agents; Area; Astrocytes; Behavioral; Brain; Cell Aging; Cell Proliferation; Cell Separation; Cells; Cognition; Data; Data Set; Department chair; Development; Disease; Environment; Epilepsy; Exercise; Exposure to; Feedback; Fluorescence-Activated Cell Sorting; Future; Goals; Grant; Growth; Health; Hippocampus (Brain); Human; Individual; Injury; Institutes; Instruction; International; Investigation; Kainic Acid; Lead; Learning; Left; Light; LoxP-flanked allele; Manuscripts; Medical; Memory; Mentors; Modeling; Motivation; Mus; Natural regeneration; Neurology; Neurons; Neurophysiology - biologic function; Occupations; Output; Pathology; Phase; Physiological; Population; Positioning Attribute; Postdoctoral Fellow; Preparation; Procedures; Process; Production; Proteins; Publications; RNA; Recovery; Regulation; Reporting; Research; Research Design; Resistance; Role; Running; Scientist; Secure; Seizures; Severities; Shapes; Single Seizures; Stem cells; Stimulus; Techniques; Technology; Testing; Training; Transgenic Mice; Undifferentiated; Universities; Vascular Endothelial Growth Factors; Vascularization; Work; Writing; adult neurogenesis; base; brain health; career; cell growth; cognitive enhancement; deep sequencing; dentate gyrus; effective therapy; experience; immunohistochemical markers; improved; injury and repair; knock-down; meetings; mouse model; nerve stem cell; nestin protein; neurogenesis; novel; prevent; programs; regenerative; relating to nervous system; research study; response; response to injury; senior faculty; stem; symposium; tenure track; therapeutic development; therapeutic target; therapy design; transcriptome sequencing

 DESCRIPTION (provided by applicant): Candidate: The candidate is a 3rd year postdoctoral fellow at Stanford University in the lab of Dr. Tony Wyss- Coray. The candidate's immediate goals are to gain independence and establish her own lab at an academic research-focused university or institute studying the role of undifferentiated neural stem or progenitor cells in hippocampal function. Her long-term goals are to build a prominent lab in the area of hippocampal plasticity and injury response at an academic research center. She aims to improve our understanding of how the hippocampus, an essential brain area for memory function, responds to the environment, either positively or negatively and thereby inform development of therapeutics for human brain health. She also plans to make high quality mentoring of young scientists a priority in her career. Environment: The proposed Mentored Phase research will be conducted in the lab of Dr. Tony Wyss-Coray at Stanford University. Co-mentors Dr. Theo Palmer and Dr. Tom Rando will also advise the candidate through quarterly individual meetings and yearly meetings of the candidate and the entire mentoring team. The Stanford Neurology department will hold yearly mandatory meetings where the candidate will present her research and career progress to senior faculty, along with other K99 awardees, and receive feedback and further guidance. Coursework at Stanford and Cold Spring Harbor Lab will provide formal instruction in lab management, grant writing and mentoring. A team of several consultants is also in place to help with technical training on several new procedures that the candidate will learn in the Mentored Phase, as well as study design using these new procedures. The transition to independence has defined milestones and will be tangibly supported by the primary mentor and co-mentors in the form of job opportunity referrals, practice with job talks, and advice on offers. The primary mentor will invest significant effort in the establishment of the candidate's lab by: 1) working with the neurology department chair to secure a tenure-track position for the candidate, 2) providing feedback on R01 preparation during the Mentored Phase and 3) promoting the candidate's new lab at national and international conferences. Research: The proposed experiments will investigate a novel functional role for undifferentiated neural stem and progenitor cells (NSPCs) in the adult brain after enhancement and injury. The proliferation of endogenous NSPCs in the adult hippocampus increases dramatically after both beneficial voluntary exercise and injurious seizures. Most previous studies have debated the function of the immature neurons that result from the differentiated products of these NSPCs. This proposal will determine the direct role of undifferentiated NSPCs in hippocampal function via secretion of growth factors after both seizures and exercise. A key motivation for this research is to inform future therapeutic targeting of NSPCs by determining how this important, unique cell population alters injury and plasticity responses in the adult hippocampus. This proposal will introduce functional implications for adult neurogenesis at a cellular age previously thought to be silent. Given the numerous environmental stimuli that impact NSPC proliferation, these findings could have wide-reaching implications for the role of NSPCs in hippocampal plasticity. The first aim of this project will characterize the growth factor response of NSPCs to physiological and pathological stimuli. Fluorescence-activated cell sorting techniques from acutely dissected murine adult hippocampus will be employed to isolate NSPCs after seizures or voluntary wheel running exercise. RNAseq will be used to contrast the profiles of NSPC transcriptional responses to pathological vs physiological stimuli. These findings will represent a rich dataset that will infor many future investigations of how NSPCs participate in regulating hippocampal function. To determine the direct role of NSPC-secreted growth factors, this proposal will use a novel mouse model for inducible, NSPC-specific knockdown of floxed growth factors, starting with an established model for inducible knockdown of the growth factor VEGF in NSPCs. For exercise, knockdown of a growth factor (like VEGF) in NSPCs will be induced in adulthood, followed by exposure to an unlocked running wheel. Behavioral and immunohistochemical markers of exercise-related enhancement of hippocampal function will then be quantified to determine whether loss of that growth factor from NSPCs prevents any benefits of exercise. For seizures, after growth factor knockdown, mice will be exposed to kainic acid either systemically or centrally to induce different levels of seizure severity. Behavioral and structural sequela will thn be quantified to determine whether loss of NSPC-growth factor is beneficial or detrimental to recovery from seizure. These studies will shed light on the function of undifferentiated neural progenitors, a largely uninvestigated cell population but one that may have important implications for design of therapies for disorders that differentially impact the hippocampus such as epilepsy.

 描述（由适用提供）：候选人：候选人是斯坦福大学托尼·韦斯斯·科雷（Tony Wyssscoray）实验室的斯坦福大学（Stanford University）的第三年。候选人的直接目标是获得独立性，并在以学术研究为中心的大学或研究所建立自己的实验室，以研究不受影化的神经茎或祖细胞在海马功能中的作用。她的长期目标是在学术研究中心在海马可塑性和伤害反应领域建立一个著名的实验室。她的目的是提高我们对海马（用于记忆功能的重要大脑领域）如何对环境做出积极或负面的反应，从而为人类脑健康理论发展提供了依据。她还计划使年轻科学家的高质量心态在职业生涯中成为重中之重。环境：拟议的指导阶段研究将在斯坦福大学的Tony Wyss-Coray博士的实验室进行。联合官员西奥·帕尔默（Theo Palmer）博士和汤姆·兰多（Tom Rando）博士还将通过季度的个人会议和候选人的年度会议和整个心理团队的年度会议建议候选人。斯坦福大学神经病学部将举行年度强制会议，候选人将与其他K99获奖者一起向高级教师提供研究和职业进步，并获得反馈和进一步的指导。斯坦福大学和冷春港实验室的课程将提供实验室管理，赠款写作和指导的正式指导。还建立了一支由几个顾问组成的团队，以帮助候选人将在指导阶段学习的几个新程序以及使用这些新程序的研究设计进行技术培训。向独立的过渡已经定义了里程碑，并将以工作机会转介的形式得到主要导师和联席会员的明显支持，练习工作谈话以及有关报价的建议。主要导师将向 建立候选人实验室的作者：1）与神经病学系主任合作确保候选人的终身任期职位，2）在指导阶段提供有关R01准备的反馈，以及3）在国家和国际会议上促进候选人的新实验室。研究：拟议的实验将研究成人大脑在增强和损伤后在成人大脑中未分化的神经元干细胞（NSPC）的新功能作用。有益的自愿运动和有害性癫痫发作后，成年海马中内源性NSPC的扩散会大大增加。以前的大多数研究都辩论了由这些NSPC的分化产物引起的未成熟神经元的功能。该建议将通过分泌癫痫发作和运动后的分泌生长因子来确定统一的NSPC在海马功能中的直接作用。这项研究的关键动机是告知未来的治疗目标 通过确定这种重要的，独特的细胞种群如何改变成人海马的损伤和可塑性反应，来实现NSPC的形象。该提案将在以前被认为是沉默的细胞时代对成人神经发生引起功能意义。鉴于影响NSPC增殖的众多环境刺激，这些发现可能对NSPC在海马可塑性中的作用具有广泛的影响。该项目的第一个目的是表征NSPC对生理和病理刺激的生长因子反应。急性解剖的鼠成年海马的荧光激活的细胞分选技术将被雇用在癫痫发作或自愿车轮运行运动后分离NSPC。 RNASEQ将用于对比NSPC转录反应对病理与生理刺激的特征。这些发现将代表一个丰富的数据集，该数据集将进一步了解NSPC如何参与确定海马功能的许多研究。为了确定NSPC分泌的生长因子的直接作用，该提案将使用新型的小鼠模型来诱导，NSPC特异性敲低生长因子，从已建立的NSPC中生长因子VEGF的诱导型敲低模型开始。为了进行运动，将在成年后诱发NSPC中生长因子（如VEGF）的敲低，然后暴露于解锁的跑步轮。然后，将量化与运动相关增强的行为相关增强的行为和免疫组织化学标志物，以确定NSPC中该生长因子的损失是否阻止了运动的任何好处。对于癫痫发作，在生长因子敲低后，小鼠将在全身或集中暴露于海藻酸中，以诱导不同水平的癫痫发作严重程度。将对行为和结构后遗症进行量化，以确定NSPC-GROWTH因子的丧失是否对癫痫发作的恢复有益或有害。这些研究将揭示未分化的神经元祖细胞的功能，神经元祖细胞（在很大程度上没有研究的细胞群体），但可能对疾病的设计具有重要意义，这些疾病的疗法对海马（如癫痫病）有差异化。