Cellular Commitment To Mortality

细胞对死亡率的承诺

• 批准号: 6508446
• 负责人: Minoru S Ko
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6508446
• 关键词: cell differentiation; cell senescence; cell transformation; early embryonic stage; embryo /fetus; embryonic stem cell; germ cells; laboratory mouse; mammalian embryology

The major goal is to understand the fundamental mechanisms for cellular commitment to mortality. We concentrate on the critical distinction between immortal early embryonic cells and mortal differentiating derivative cells. Our approach is to optimize the use of systematic genomic approaches for profiling gene expression patterns by large-scale cDNA sequencing and cDNA microarray technology with a current cohort of 15,257 genes collected from early mouse embryos. We are particularly focusing on two closely related experimental systems. (1) Preimplantation development - the process of fertilized egg gradually losing its totipotency. In our previous work, we identified many genes that show stage-specific expression patterns during preimplantation mouse development. However, these genes have been identified by EST frequency, which is a relatively inaccurate and far from ideal way to do gene expression profiling. The cDNA microarray-based gene expression profiling will provide more reliable information. To this end, we have been working on large-scale gene expression profiling of each stage of preimplantation mouse development. To start to examine whether these genes identified here might be involved in the immortal/mortal transition process, we are currently analyzing the nature of these genes in greater detail. We are also testing various antisense oligonucleotides to inhibit the function of specific gene in preimplantation mouse embryos. (2) Differentiation of the urogenital system: Germ line cells are often viewed as immortal, because they provide continuity from generation to generation and do not seem to age. To begin the analysis of gene expression regulation involved in germ line-somatic cell interactions, we conducted gene expression profiling by sequencing cell type-specific cDNA libraries that include E13.5 male and female primordial germ cells as well as two stages in the kidney-urogenital developmental axis, E12.5 female mesonephros and newborn ovary. We are currently analyzing genes differentially expressed in these cells, tissues, and organs more in detail.

主要目标是了解细胞死亡的基本机制。我们集中在不朽的早期胚胎细胞和致命的分化衍生细胞之间的关键区别。我们的方法是通过大规模cDNA测序和cDNA微阵列技术优化使用系统基因组方法来分析基因表达模式，目前从早期小鼠胚胎中收集了15，257个基因。我们特别关注两个密切相关的实验系统。(1)着床前发育-受精卵逐渐失去全能性的过程。在我们以前的工作中，我们确定了许多基因，在植入前小鼠发育过程中表现出阶段特异性表达模式。然而，这些基因已被确定的EST频率，这是一个相对不准确的，远远不够理想的方式做基因表达谱。基于cDNA微阵列的基因表达谱分析将提供更可靠的信息。为此，我们一直致力于对植入前小鼠发育的每个阶段进行大规模基因表达谱分析。为了开始研究这些基因是否参与了不朽/凡人的转变过程，我们目前正在更详细地分析这些基因的性质。我们也正在测试各种反义寡核苷酸来抑制特定基因在植入前小鼠胚胎中的功能。(2)泌尿生殖系统的分化：生殖系细胞通常被认为是不朽的，因为它们提供了一代又一代的连续性，并且似乎不会老化。为了开始分析生殖系-体细胞相互作用中涉及的基因表达调控，我们通过对细胞类型特异性cDNA文库进行测序来进行基因表达谱分析，所述cDNA文库包括E13.5雄性和雌性原始生殖细胞以及肾脏-泌尿生殖器发育轴中的两个阶段，E12.5雌性中肾和新生儿卵巢。我们目前正在更详细地分析这些细胞、组织和器官中差异表达的基因。