Acquisition of confocal scan to upgrade existing fluorescence microscope

获取共焦扫描以升级现有荧光显微镜

• 批准号: 10796239
• 负责人: Emilia Galperin
• 金额: $ 23.45万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10796239
• 关键词: Address; Administrative Supplement; Biological Markers; Cells; Complex; Development; Disease; Embryonic Development; Endosomes; Equipment; Fostering; Future; Genes; Image; MAPK3 gene; Malignant Neoplasms; Mediating; Microscope; Modeling; Molecular; Outcome; Pathway interactions; Research; Role; Scaffolding Protein; Scanning; Signal Transduction; Sorting; Specificity; Syndrome; Ubiquitin; Work; Zebrafish; developmental disease; experimental study; fluorescence microscope; improved; in vivo; insight; novel diagnostics; novel therapeutic intervention; programs; public health relevance; segregation; targeted treatment; therapeutic development; transmission process; tumor; virtual

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.

项目摘要 本提案是对现有MIRA R35的设备管理补充，以支持PI 细胞外信号调节激酶1和2（ERK 1/2）的分子机制研究 由支架蛋白引导的信号传输。该研究计划的重点是了解 关键调节因子如支架蛋白Shoc 2修饰ERK 1/2信号传导的机制。这些 研究破译并确定了内体分选调节泛素驱动的 Shoc 2复合物的重塑和信号传导，并阐明了将Shoc 2分离成不同的 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 Shoc 2激活的ERK 1/2信号如何促进早期发育阶段。在这项工作中，斑马鱼被用来 作为脊椎动物模型来研究Shoc 2基因编辑对早期阶段的体内影响 因为它们为发展研究提供了独特的优势，包括其透明度， 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， 对Shoc 2如何参与决定ERK 1/2特异性的全面机制理解 信号结果。因此，这项工作为未来对发育障碍的研究奠定了基础 这将有助于新的治疗策略和生物标志物的发展。这一行政 作为现有3 i Marianas的“附加”，获得共焦扫描盘的补充应用 显微镜平台将促进这项研究计划正在进行的实验。高级扫描 能力将显着提高成像分析的质量，以检查Shoc 2的分布 细胞中的信号复合物，并将大大加强这些研究，以检查Shoc 2的作用， 胚胎发育中的信号。