NEURONAL TURNOVER IN ADULT BRAIN USING AMS FOR RETROSPECTIVE BIRTH DATING CELLS

使用 AMS 进行追溯出生约会细胞的成人大脑神经元周转

• 批准号: 7602409
• 负责人: JONAS FRISEN
• 金额: $ 2.85万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602409
• 关键词: Adult; Affect; Age; Authorization documentation; Autopsy; Biological Process; Blood; Brain; C14 isotope; Carbon; Cell Aging; Cell division; Cells; Computer Retrieval of Information on Scientific Projects Database; Condition; DNA; Date of birth; Disease; Drops; Equus caballus; Funding; Generations; Genomics; Grant; Human; Human Pathology; Individual; Institution; Isotopes; Measurement; Measures; Methods; Neuroglia; Neurons; Nuclear; Pathologic Processes; Physiologic pulse; Physiological; Population; Pulse taking; Range; Relative (related person); Research; Research Personnel; Resolution; Resources; Sorting - Cell Movement; Source; Techniques; Testing; Thinking; Time; Tooth structure; Tubulin; United States National Institutes of Health; base; cell age; insight; interest; man; mass spectrometer; mature animal; neurogenesis; novel; novel therapeutics; planetary Atmosphere; postnatal; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Neurogenesis is known to occur in specific regions of the adult animal brain, but the extent and comparability of neurogenesis in the adult human brain is much harder to determine, and to date largely unknown. Traditional methods used for dating cells are limited in the information they provide, or are not appropriate for human use. Thus, currently there is no method available to study cellular turnover in man. We propose to develop a method for the retrospective birth dating of cells. We are interested in using bomb pulse carbon-14 (C14) dating as a method for measuring the approximate age of specific populations of cells in the adult human brain. This method is based on establishing the proportion of the isotope C14 in genomic DNA. C14 measurements will be made using the highly sensitive accelerator mass spectrometer (AMS). After a cell has terminally differentiated it does not divide again. Since the last cell division represents the last time point when the cell synthesized DNA, its chromosomal DNA will reflect the age when the cell was born. Traditionally, the slow decay of C14 relative to other carbon species has given it a temporal resolution of many years, however due to nuclear tests in the late 1950s and early 1960s, the level of C14 in the atmosphere has increased dramatically. This level has since dropped off in an exponential fashion, allowing one to resolve C14 differences in the range of years. Because DNA has a C14 content reflective of the time when it was synthesized, establishing the C14 content of chromosomal DNA will enable us to retrospectively birth date cells, and thus establish cellular turnover. Crucial to the understanding of basic biological processes, is information about cellular turnover. As well as having an interest in normal cellular turnover, many diseases are thought to be affected in their generation of new cells. Information about cellular turnover in disease states may provide novel insights into the pathological processes of the disease, and possibly suggest new therapeutic strategies. Preliminary experiments using AMS to date C14 from horse brain DNA have yielded encouraging results. The next step is to look at how accurately AMS dates C14 from DNA extracted from horse blood, brain and teeth. Particular populations of cells will then be isolated using FACS analysis (which allows specific cell populations to be isolated e.g. one can sort for neurons using neuronal specific markers such as NeuN, ¿ tubulin or Thy1). Once this technique has been established we aim to move onto human material, and study cellular turnover in specific human pathologies. Ethical permission to obtain postmortem human material has been granted. We used this strategy to determine the age of cells in the cortex of the adult human brain, and showed that whereas non-neuronal cells are exchanged, occipital neurons are as old as the individual, supporting the view that postnatal neurogenesis does not take place in this region. Retrospective birth dating is a generally applicable strategy that can be used to measure cell turnover in man under physiological and pathological conditions.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 已知神经发生发生在成年动物大脑的特定区域，但神经发生的程度和可比性 成人大脑中的神经发生很难确定，迄今为止，传统方法基本上未知。 用于测定细胞的信息有限，或者不适合人类使用。 没有可用于研究人类细胞更新的方法。我们建议开发一种回顾性方法。 我们有兴趣使用炸弹脉冲碳 14 (C14) 测年作为测量细胞诞生的方法。 该方法基于建立成人大脑中特定细胞群的大致年龄。 基因组 DNA 中同位素 C14 的比例将使用高灵敏度加速器进行测量。 质谱仪（AMS）。 细胞最终分化后不会再次分裂，因为最后一次细胞分裂代表最后一次。 当细胞合成DNA时，其染色体DNA将反映细胞出生时的年龄。 C14 相对于其他碳物种的缓慢衰变使其具有多年的时间分辨率，但是由于 20世纪50年代末和60年代初的核试验导致大气中C14的含量急剧增加。 此后，C14 水平呈指数下降，从而可以解决年份范围内的 C14 差异。 由于 DNA 的 C14 含量反映了其合成时间，因此建立了 染色体 DNA 将使我们能够追溯细胞的出生日期，从而建立细胞更替。 对于理解基本生物过程至关重要的是有关细胞更新的信息。 由于对正常细胞周转的兴趣，许多疾病被认为会受到新细胞生成的影响。 有关疾病状态下细胞更新的信息可能为了解疾病的病理过程提供新的见解。 疾病，并可能提出使用 AMS 测定马 C14 的初步实验。 大脑 DNA 已经取得了令人鼓舞的结果，下一步是研究 AMS 如何准确地从 DNA 中确定 C14。 然后使用 FACS 分析从马的血液、大脑和牙齿中提取特定的细胞群。 （这允许分离特定的细胞群，例如可以使用神经元特异性标记物对神经元进行分类 作为 NeuN, ¿一旦这项技术建立起来，我们的目标就是转移到人体材料上，并且 研究特定人类病理学中的细胞更新已获得道德许可以获取死后人类材料。 被授予。 我们使用这种策略来确定成人大脑皮层中细胞的年龄，并表明，而 非神经元细胞被交换，枕叶神经元与个体一样老，支持出生后的观点 神经发生不发生在该区域，是一种普遍适用的策略。 用于测量人体在生理和病理条件下的细胞更新。