Germline Stem Cell Analysis Tools

 描述（由申请人提供）：本项目将开发，优化和实施技术，以线性标记和/或遗传改变单个生殖细胞在C。优雅这种蠕虫的生殖系与包括人类在内的所有动物的干细胞系统有许多共同特征。例如，它通过与细胞微环境或小生境的相互作用来维持，所述细胞微环境或小生境产生配体以激活生殖细胞上的高度保守的Notch受体。这是第一个在细胞和分子水平上定义小生境的干细胞系统。这个干细胞系统已经成为一个强大的系统，以了解干细胞扩增，静止和可塑性的动态响应遗传，环境和生理变化。蠕虫的生命周期很短（3天），是透明的，短暂的，并且非常适合环境和遗传操纵。尽管有这些优势，了解生殖系干细胞系统的关键特征及其对环境和遗传扰动的反应目前受到我们对细胞更新机制以及它们如何在实验操作下改变的相对较差的理解的限制。为了弥合这一差距，我们需要随着时间的推移跟踪单个干细胞及其细胞后代。这里提出的技术将弥合这一差距。利用近年来在转基因表达方面的研究进展，elegans，现有的技术从其他生物体将被带到蠕虫。除了谱系标记之外，该技术还将使操纵单细胞或细胞亚群中的基因活性成为可能。该模型系统的独特之处在于，该技术将能够在整个生命周期和整个动物生理扰动的条件下监测个体的生殖干细胞。这一进展将有利于更大的干细胞生物学和生殖系领域的其他问题将成为在这个总理模型系统访问。

 DESCRIPTION (provided by applicant): This project will develop, optimize and implement technology to lineally mark and/or genetically alter individual germ cells in C. elegans. The germ line in this worm shares many features in common with stem cell systems in all animals, including humans. For example, it is maintained by virtue of interaction with a cellular microenvironment or niche that produces ligands to activate a highly conserved Notch receptor on the germ cells. It was the first stem cell system for which a niche was defined at the cell and molecular levels. This stem cell system has since emerged as a powerful system to understand the dynamics of stem cell expansion, quiescence and plasticity in response to genetic, environmental and physiological alterations. The worm has a short (3-day) lifecycle, is transparent, short-lived, and highly amenable to environmental and genetic manipulation. Despite these advantages, understanding critical features of the germline stem cell system and its responses to environmental and genetic perturbations is currently limited by our relatively poor understanding of cellular mechanisms of renewal and how they are altered under experimental manipulation. To bridge this gap, we need to follow individual stem cells and their cellular progeny over time. The technology proposed here will bridge this gap. Taking advantage of recent progress in germline transgene expression in C. elegans, existing technologies from other organisms will be brought to the worm. In addition to lineage marking, the technology will make possible the manipulation of gene activity in single cells or subsets of cells. Uniquely for this model system, the technology will enable monitoring of germline stem cells in individuals over their entire lifetime and in conditions of whole-animal physiological perturbations. This advance will benefit the greater stem cell biology and germline fields as additional questions will become accessible in this premier model system.