The role of dynamics in defining the limits of normal developmental signaling

动力学在定义正常发育信号限制中的作用

• 批准号: 10390229
• 负责人: John G. Albeck
• 金额: $ 19.39万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390229
• 关键词: Affect; Aging; Automation; Autophagosome; Biosensor; Cancer Biology; Cell Line; Cell Proliferation; Cell model; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collecting Cell; Communication; Computer Models; Data; Data Set; Defect; Development; Dose; Equipment; Event; Face; Failure; Fos-Related Antigens; Funding; Gene Expression; Gene Expression Process; Gene Expression Profile; Genes; Genetic Transcription; Germ-Line Mutation; Grant; Growth Factor; Heart Abnormalities; Homeostasis; Human; Human Resources; Hyperactivity; Image; Imaging Techniques; Immersion; Impaired cognition; Kinetics; Knock-in; Lead; Link; Live Birth; Malignant Neoplasms; Measurement; Methodology; Methods; Microscopy; Mitochondria; Modeling; Molecular; Morphology; Mutation; Nature; Organelles; Pathologic; Pathology; Pathway interactions; Pattern; Pharmacology; Phenotype; Phosphotransferases; Probability; Process; Proteins; Regulation; Reporter; Resolution; Role; Running; Series; Signal Transduction; Signal Transduction Pathway; Surveys; Syndrome; System; Technology; Testing; Time; Tissues; Translations; Variant; Water; Work; cancer risk; cell behavior; cell motility; cognitive development; developmental disease; experimental study; fluorescence microscope; genetic regulatory protein; human disease; imaging system; improved; individualized medicine; inhibitor/antagonist; instrument; lens; live cell microscopy; mRNA Differential Displays; mutant; parent project; preference; response; single cell analysis; temporal measurement; transmission process

Summary In this supplement, we request funding to purchase a live-cell fluorescence microscope that will replace failing essential equipment, and which will significantly upgrade our ability to collect cell signaling biosensor data with high spatial and temporal resolution. The parent project for this supplement is centered on using live-cell biosensor experiments to collect kinetic data on kinase activity and gene expression. These datasets uniquely enable computational modeling of the relationship between signal transduction pathways and gene expression. Because of the time-intensive nature of these long-term time-lapse imaging experiments, we need a dedicated live-cell microscopy system that can run continuously for multi-day experiments. Our current systems have enabled significant conceptual and methodological advances in growth factor signaling but face two major problems: 1) frequent down-time due to failures of aging equipment and 2) outdated technology that limits our ability to collect high-quality data from live cells. The proposed instrument would dramatically decrease the amount of personnel time lost to component failures. Furthermore, the proposed instrument will take advantage of a large field of view sCMOS camera that can collect data from nearly twice as many cells in each experiment; these additional data will directly improve our computational modeling efforts by capturing more unique cellular behaviors. Finally, the new instrument will include automation to enable the use of 40X and 60X water immersion objective lenses for long-term time time-lapse experiments. Relative to our current imaging systems that rely on 20X and 40X non-immersion lenses, these lenses will increase spatial resolution and sensitivity, enabling experiments in which we can link signal transduction and gene expression to morphological changes in key organelles such as autophagosomes and mitochondria, which our work has implicated in regulation of the dynamic kinase activity being studied.

概括 在本增刊中，我们请求资金购买活细胞荧光显微镜，以取代失败的产品 必要的设备，这将显着提高我们收集细胞信号生物传感器数据的能力 高空间和时间分辨率。本补充品的父项目以使用活细胞为中心 生物传感器实验收集激酶活性和基因表达的动力学数据。这些数据集独一无二 能够对信号转导途径和基因表达之间的关系进行计算建模。 由于这些长期延时成像实验的时间密集性，我们需要一个专用的 活细胞显微镜系统，可以连续运行进行多天的实验。我们当前的系统有 使生长因子信号传导在概念和方法上取得了重大进展，但面临两个主要问题 问题：1）由于设备老化故障而频繁停机；2）过时的技术限制了我们的 从活细胞收集高质量数据的能力。拟议的文书将大大减少 因组件故障而损失的人员时间。此外，拟议的文书将利用 大视场 sCMOS 相机可以在每次实验中从近两倍数量的细胞收集数据； 这些额外的数据将通过捕获更多独特的细胞来直接改进我们的计算建模工作 行为。最后，新仪器将包括自动化功能，可使用 40X 和 60X 水浸式 用于长期延时实验的物镜。相对于我们目前依赖的成像系统 20X 和 40X 非浸没式镜头，这些镜头将提高空间分辨率和灵敏度，从而使 我们可以将信号转导和基因表达与关键的形态变化联系起来的实验 我们的工作涉及自噬体和线粒体等细胞器的调节 正在研究动态激酶活性。

项目成果

Microenvironmental Signals and Biochemical Information Processing: Cooperative Determinants of Intratumoral Plasticity and Heterogeneity.

微环境信号和生化信息处理：瘤内可塑性和异质性的协同决定因素。

• 发表时间: 2018
• 作者: Davies, Alexander E;Albeck, John G
• 通讯作者: Albeck, John G

ERK signaling dynamics: Lights, camera, transduction.

ERK 信号动力学：灯、摄像头、传导。

• 发表时间: 2022-09-26
• 影响因子: 11.8
• 作者: Ram, Abhineet;Albeck, John G
• 通讯作者: Albeck, John G

Experimental and engineering approaches to intracellular communication.

细胞内通讯的实验和工程方法。

• DOI: 10.1042/ebc20180024
• 发表时间: 2018-10-26
• 期刊: Essays in biochemistry
• 作者: Albeck JG;Pargett M;Davies AE
• 通讯作者: Davies AE