Application of Transient Intervention and Live-Imaging Methods to Resolve Cellular and Molecular Networks Regulating Stem-Cell Homeostatis in Arabidopsis Shoot Apex

Venugopala Reddy Gonehal0718046应用瞬时干预和活体成像方法来解决细胞和分子网络调节拟南芥茎尖干细胞稳态的问题干细胞的顶端细胞分裂对于确保动植物组织和器官的适当发育、维护和修复至关重要。在植物中，茎尖(顶端分生组织；SAM)含有一组干细胞，这些干细胞被分化细胞包围。干细胞的数量保持不变，即使干细胞的后代不断地被转移到分化细胞。因此，SAM代表了不同细胞类型的动态网络，其中生长和分化通过精确控制基因表达的空间和时间模式的细胞-细胞通信网络来动态调节。为了理解细胞如何解释这种复杂的信号混合物来调节细胞分裂，需要开发新的方法来识别网络组件并阐明它们的调控动力学和功能。在这项提案中，PI将利用实时成像技术应用瞬时遗传和物理操作来探索干细胞维持的关键调节因子的功能和调节。PI还将结合瞬时基因操作和干细胞浓缩方法与微阵列技术，以开发活跃生长的SAM中细胞和分子相互作用的动态图谱，最终目标是开发调节干细胞动态平衡的细胞-细胞通信预测模型。将为博士后、研究生和本科生提供设计瞬时基因操作、表情图谱和实时成像的新方法的培训。他们将接触到最先进的成像系统和图像处理平台，用于可视化、渲染和分析微观数据。让新一代科学家尽早接触到动态的数据采集和数据处理方法，将使他们为未来的定量成像挑战做好准备。

Venugopala Reddy Gonehal0718046Application of transient intervention and live-imaging methods to resolve cellular and molecular networks regulating stem-cell homeostasis in Arabidopsis shoot apex Cell division of stem-cells is critical to ensure proper development, maintenance and repair of tissues and organs of both animals and plants. In plants, the shoot tip (shoot apical meristem; SAM) harbors a set of stem-cells that are surrounded by differentiating cells. The stem-cell number remains constant, even though the stem-cell progeny are being continually diverted to differentiating cells. Thus, the SAM represents a dynamic network of different cell types in which growth and differentiation are dynamically regulated through a cell-cell communication network that precisely controls spatial and temporal patterns of gene expression. Understanding how cells interpret this complex mixture of signals to regulate cell division requires the development of new methods that identify network components and elucidate their regulatory dynamics and function. In this proposal, the PI will apply transient genetic and physical manipulations with live-imaging technologies to explore the function and regulation of key regulators of stem-cell maintenance. The PI also will combine transient gene manipulation and stem-cell enrichment methods with microarray technology to develop a dynamic map of cellular and molecular interactions in actively growing SAMs with the ultimate objective of developing predictive models of cell-cell communication that mediate stem cell homeostasis. Training will be provided for a post-doctoral fellow, graduate student and under-graduate students in designing new methods of transient gene manipulation, expression profiling and live-imaging. They will be exposed to state-of-the art imaging systems and image processing platforms for visualization, rendering and analysis of microscopic data. Early exposure of a new generation of scientists to dynamic methods of data acquisition and data handling would prepare them for future challenges of quantitative imaging.