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: 8362753
• 负责人: JONAS FRISEN
• 金额: $ 11.7万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362753
• 关键词: Adipocytes; Adult; Affect; Age; Biological Process; Brain; C14 isotope; Carbon; Cardiac Myocytes; Cell division; Cells; DNA; Date of birth; Dental Enamel; Disease; Drops; Forensic Medicine; Funding; Generations; Genomics; Grant; Human; Investigation; Isotopes; Lipids; Measurement; Measures; Methods; National Center for Research Resources; Neurons; Nuclear; Pathologic Processes; Physiologic pulse; Population; Principal Investigator; Relative (related person); Research; Research Infrastructure; Resolution; Resource Development; Resources; Sorting - Cell Movement; Source; Techniques; Testing; Time; United States National Institutes of Health; accelerator mass spectrometry; base; cost; insight; interest; man; mass spectrometer; mature animal; neurogenesis; novel; novel therapeutics; planetary Atmosphere; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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. 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). The technique has also been applied to cardiomyocytes and adipocytes. We are investigating the turnover of lipids within adipocytes also. The use of enamel as a forensic tool to establish date of birth has been under investigation the past couple years.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 已知神经发生发生在成年动物大脑的特定区域，但成年人类大脑中神经发生的程度和可比性更难确定，到目前为止基本上是未知的。用于细胞测年的传统方法提供的信息有限，或者不适合人类使用。因此，目前还没有可用的方法来研究人类的细胞周转。我们建议开发一种细胞追溯出生年龄的方法。我们感兴趣的是使用核弹脉冲碳14(C14)测年作为一种测量成人大脑中特定细胞群体的近似年龄的方法。该方法建立在确定同位素C14在基因组DNA中的比例的基础上。C14的测量将使用高灵敏度的加速器质谱仪(AMS)进行。 细胞终末分化后，不会再分裂。由于最后一次细胞分裂代表细胞合成DNA的最后时间点，其染色体DNA将反映细胞出生的年龄。传统上，C14相对于其他碳物种的缓慢衰减使其具有多年的时间分辨率，然而，由于20世纪50年代末和60年代初的核试验，大气中C14的水平急剧上升。自那以后，这一水平以指数方式下降，使人们能够在几年的范围内解决C14的差异。因为DNA的C14含量反映了它合成的时间，所以确定染色体DNA的C14含量将使我们能够追溯出生日期细胞，从而建立细胞周转。 细胞周转的信息对于理解基本的生物过程至关重要。除了对正常的细胞周转感兴趣外，许多疾病被认为是在新细胞的生成过程中受到影响。有关疾病状态下细胞周转的信息可能为疾病的病理过程提供新的见解，并可能提出新的治疗策略。然后，将使用FACS分析来分离特定的细胞群体(这允许分离特定的细胞群体，例如，可以使用神经元特异性标记，如NeuN)来分选神经元。这项技术也被应用于心肌细胞和脂肪细胞。我们也在研究脂肪细胞内脂肪的周转。在过去的几年里，人们一直在调查牙釉质作为确定出生日期的法医工具的问题。