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: 8171680
• 负责人: JONAS FRISEN
• 金额: $ 3.4万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171680
• 关键词: Adult; Affect; Age; Authorization documentation; Autopsy; Biological Process; Blood; Brain; C14 isotope; Carbon; Cell division; Cells; Computer Retrieval of Information on Scientific Projects Database; DNA; Date of birth; Disease; Drops; Equus caballus; Ethics; Funding; Generations; Genomics; Grant; Human; Human Pathology; Institution; Isotopes; Measurement; Measures; Methods; Neurons; Nuclear; Pathologic Processes; Physiologic pulse; Population; Relative (related person); Research; Research Personnel; Resolution; Resources; Sorting - Cell Movement; Source; Techniques; Testing; Time; Tooth structure; Tubulin; United States National Institutes of Health; base; insight; interest; man; mass spectrometer; mature animal; neurogenesis; novel; novel therapeutics; planetary Atmosphere; 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.

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。 中心，不一定是研究者的机构。 众所周知，神经发生发生在成年动物大脑的特定区域，但成人大脑中神经发生的程度和可比性更难确定，迄今为止，用于测定细胞年代的传统方法在信息方面还很有限。提供或不适合人类使用因此，目前没有可用于研究人类细胞更新的方法，我们对使用炸弹脉冲碳14感兴趣。 C14) 约会作为测量近似值的方法该方法基于使用高灵敏度加速器质谱仪 (AMS) 确定基因组 DNA 中同位素 C14 的比例。 细胞最终分化后不会再次分裂，由于最后一次细胞分裂代表细胞合成DNA的最后一个时间点，因此其染色体DNA将反映细胞出生时的年龄。传统上，C14相对于细胞的缓慢衰退。其他碳物种使其具有多年的时间分辨率，但由于 20 世纪 50 年代末和 1960 年代初的核试验，大气中的 C14 含量急剧增加，此后该含量已急剧下降。指数方式，允许人们解决年份范围内的 C14 差异，因为 DNA 的 C14 含量反映了它的合成时间，建立染色体 DNA 的 C14 含量将使我们能够追溯细胞的出生日期，从而建立细胞。周转。 对于理解基本生物过程至关重要的是关于细胞更新的信息以及对正常细胞更新的兴趣，许多疾病被认为在新细胞的生成中受到影响关于疾病状态下的细胞更新的信息可能提供新的信息。利用 AMS 测定马脑 DNA 中的 C14 的初步实验取得了令人鼓舞的结果，下一步是研究 AMS 从马血中提取的 DNA 中测定 C14 的准确性。 、大脑和牙齿。然后使用 FACS 分析分离特定的细胞群（这允许分离特定的细胞群，例如可以使用 NeuN、微管蛋白或 Thy1 等神经元特异性标记物对神经元进行分类）一旦建立了这项技术，我们的目标就是移动。人类材料，并研究特定人类病理学中的细胞更新，已获得获得死后人类材料的伦理许可。