Acquisition of confocal scan to upgrade existing fluorescence microscope

项目摘要 这项提议是对现有的支持PI的Mira R35的设备管理补充 细胞外信号调节蛋白1和2(ERK1/2)分子机制研究 由支架蛋白引导的信号传递。本研究计划的重点是了解 关键调节者，如支架蛋白Shoc2，修改ERK1/2信号的机制。这些 研究破译和确定内体分选调节泛素驱动的机制 Shoc2复合体和信号转导的重塑，并说明将Shoc2分离为不同的关键步骤 细胞内分选途径。初步研究已经对分子细节产生了新的见解 Shoc2激活的ERK1/2信号如何促进早期发育阶段。在这项工作中，使用了斑马鱼 作为脊椎动物模型来研究Shoc2基因编辑在早期阶段的体内效应 发展，因为它们为发展研究提供了明显的优势，包括其透明度，如 以及外部快速发展。这项工作的成果预计将产生一个雄心勃勃的 全面理解Shoc2如何参与决定ERK1/2的特异性 发出结果信号。因此，这项工作正在为未来对发育障碍的研究奠定基础 这将有助于新的治疗策略和生物标记物的发展。这项管理工作 补充申请以获取共焦扫描盘，作为现有3i Marianas的附加组件 显微镜平台将在这一研究计划中促进正在进行的实验。先进的扫描技术 能力将显著提高检查Shoc2分布的成像分析的质量 细胞内的信号复合体，并将极大地加强这些研究，以检验Shoc2- 胚胎发育中的中介信号。

Project Summary This proposal is for an equipment administrative supplement to an existing MIRA R35 supporting the PI’s research addressing the molecular mechanisms of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) signal transmission as guided by scaffold proteins. This research program focuses on understanding the mechanisms by which key regulators, such as the scaffold protein Shoc2, modify ERK1/2 signaling. These studies decipher and determine the mechanisms by which endosomal sorting regulates the ubiquitin-driven remodeling of Shoc2 complexes and signaling and illuminate the key steps that segregate Shoc2 into distinct intracellular sorting pathways. Preliminary studies have already yielded new insights into the molecular details of how Shoc2-activated ERK1/2 signals promote early developmental stages. For this work, zebrafish is used as a vertebrate model to investigate the in vivo effect of Shoc2 gene editing on the early stages of development since they offer distinct advantages for studies of development, including their transparency, as well as external and rapid development. Outcomes from this work are expected to produce an ambitious and comprehensive mechanistic understanding of how Shoc2 is involved in determining the specificity of ERK1/2 signaling outcomes. As such, this work is laying the groundwork for future studies on developmental disorders that will contribute to the advancement of novel therapeutic strategies and biomarkers. This administrative supplement application to acquire a confocal scanning disk as an ‘add-on’ to an existing 3i Marianas microscope platform will foster ongoing experiments in this research program. The advanced scanning capacities will significantly improve the quality of the imaging analysis to examine the distribution of Shoc2 signaling complexes in cells and will dramatically enhance these studies to examine the role of Shoc2- mediated signals in embryonic development.