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（由申请人提供）：候选人：候选人是斯坦福大学博士实验室的第三年博士后研究员。候选人的直接目标是获得独立性，并在以学术研究为重点的大学或研究所建立自己的实验室，研究未分化的神经干细胞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建立一个海马可塑性和损伤反应领域的著名实验室。她的目标是提高我们对海马体（记忆功能的重要大脑区域）如何对环境做出积极或消极反应的理解，从而为人类大脑健康的治疗方法的发展提供信息。她还计划在她的职业生涯中优先考虑对年轻科学家的高质量指导。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。共同导师Theo Palmer博士和Tom Rando博士还将通过候选人和整个指导团队的季度个人会议和年度会议为候选人提供建议。斯坦福大学神经病学系将举行年度强制性会议，候选人将向高级教师介绍她的研究和职业进展，沿着其他K99获奖者，并获得反馈和进一步指导。 在斯坦福大学和冷泉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示。此外，还有一个由多名顾问组成的团队，帮助候选人在辅导阶段学习的几个新程序的技术培训，以及使用这些新程序的研究设计。 向独立的过渡已经确定了里程碑，并将得到主要导师和共同导师的切实支持，其形式是工作机会推荐、工作会谈实践和提供建议。主要导师将投入大量精力， 通过以下方式建立候选人的实验室：1）与神经病学系主任合作，确保候选人的终身职位，2）在指导阶段提供关于R 01准备的反馈，3）在国家和国际会议上宣传候选人的新实验室。研究：本实验将研究未分化神经干细胞和祖细胞在成年脑增强和损伤后的新功能作用。成年海马内源性NSPCs的增殖在有益的自主运动和损伤性癫痫发作后均显著增加。大多数以前的研究争论的功能，未成熟的神经元，从这些NSPCs的分化产物。该提议将确定癫痫发作和运动后未分化的NSPCs通过分泌生长因子在海马功能中的直接作用。这项研究的一个关键动机是为未来的治疗靶向提供信息 通过确定这种重要的，独特的细胞群如何改变成年海马的损伤和可塑性反应， 这项建议将介绍功能的影响，成人神经发生在细胞年龄以前被认为是沉默的。考虑到影响NSPC增殖的众多环境刺激，这些发现可能对NSPCs在海马可塑性中的作用产生广泛的影响。本项目的第一个目标是描述NSPCs对生理和病理刺激的生长因子反应。将采用来自急性解剖的成年鼠海马体的活化细胞分选技术来分离癫痫发作或自主轮跑运动后的NSPC。RNAseq将用于对比NSPC对病理性刺激与生理性刺激的转录应答谱。这些发现将代表一个丰富的数据集，将inforfor许多未来的调查如何NSPCs参与调节海马功能。 为了确定NSPC分泌的生长因子的直接作用，该提议将使用一种新的小鼠模型用于诱导型、NSPC特异性敲除floxed生长因子，从建立的用于诱导型敲除NSPC中生长因子VEGF的模型开始。对于运动，NSPCs中生长因子（如VEGF）的敲低将在成年期诱导，然后暴露于未锁定的跑步轮。然后将量化运动相关的海马功能增强的行为和免疫组织化学标志物，以确定NSPCs的生长因子损失是否会阻止运动的任何益处。对于癫痫发作，在生长因子敲低后，将小鼠全身或中枢暴露于红藻氨酸以诱导不同水平的癫痫发作严重程度。然后将量化行为和结构后遗症，以确定NSPC生长因子的丧失对癫痫发作的恢复是有益还是有害。这些研究将揭示未分化的神经祖细胞的功能，这是一个基本上未经研究的细胞群，但可能对设计差异影响海马体（如癫痫）的疾病的治疗方法具有重要意义。