Quantitative analysis of signaling dynamics across the BMP morphogen gradient

BMP 形态发生素梯度信号动力学的定量分析

基本信息

批准号：
10627788
负责人：
Susanna Elizabeth Brantley
金额：
$ 7.18万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10627788
关键词：
Address Affect Binding Biochemical Biological Assay Biological Models Biological Phenomena Bone Regeneration Cell Nucleus Cells Clinic Complex Data Development Diameter Dimerization Disease Dorsal Drops Drosophila genus Drosophila sax protein Embryo Exposure to Gene Expression Genes Genetic Genetic Transcription Goals Homeostasis Image Imaging Device In Vitro Knowledge Label Lateral Lead Ligands Ligation Maps Measures Methods Modeling Nuclear Translocation Output Pathway interactions Pattern Perception Phosphorylation Population Positioning Attribute Process Receptor Signaling Reporter Shapes Signal Pathway Signal Transduction Signal Transduction Pathway Specific qualifier value Specificity System Testing Therapeutic Time Tissue Stains Tissues Transcript Translating Visualization cohort combinatorial dosage experimental study extracellular fluorophore fly gastrulation genetic manipulation imaging modality in vivo intercellular communication morphogens rate of change receptor regenerative biology response spatiotemporal tissue fixing transcription factor transmission process

项目摘要

ABSTRACT Our current knowledge of BMP signaling pathway specificity is based upon foundational genetic and in vitro experiments, but we have limited understanding as to how cells produce diverse but specific responses to similar signaling inputs in vivo. In the Drosophila embryo, a steep gradient of BMP signaling is dynamically established prior to gastrulation. This gradient is interpreted by different populations of cells to establish the dorsal-ventral axis of the embryo, with cells at the dorsal midline turning on a unique set of transcripts compared to more lateral cells. We do not know how the dynamics of gradient formation and the final gradient pattern are interpreted by cohorts of cells in the embryo to produce the correct spatiotemporal transcriptional response. Using new imaging tools and a quantitative systems level approach, I will be able to interrogate how upstream inputs to the BMP signaling pathway are decoded in the nucleus to elicit correct spatiotemporal transcriptional responses. In this study I will explore two potential mechanisms by which BMP signaling is encoded. First, new methods will allow me to assay the signaling dynamics of pathway activity along the BMP gradient. With these data I will build an input-output relationship model to predict the mechanisms that drive BMP-responsive transcriptional dynamics. Second, I will determine if different combinations of ligand and receptor pairs play a role in specification and interpretation of the BMP gradient. I will study how both dynamic and combinatorial signaling are used to produce the wild type pattern of BMP signaling responses in the embryo. Then, using genetic perturbations in key pathway components that propagate and shape the morphogen gradient, I will test if these input-output relationships in dynamics and combinatorial signaling hold true across different BMP signaling contexts. Successful completion of this study will provide a quantitative view for how a conserved signaling pathway is interpreted by cells in vivo across time and space.

抽象的我们目前对BMP信号通路特异性的了解是基于基础遗传和体外的实验，但我们对细胞如何产生多样化但特定的反应的理解有限体内类似的信号输入。在果蝇胚胎中，BMP信号的陡峭梯度是动态的在胃之前建立。这种梯度由不同的细胞种群解释以建立胚胎的背腹轴，背部中线的细胞打开了一组独特的转录本与更多的侧细胞相比。我们不知道梯度形成的动力和最终的动力如何梯度模式由胚胎中的细胞同类解释以产生正确的时空转录响应。使用新的成像工具和定量系统级方法，我将能够询问如何在细胞核中解码到BMP信号通路的上游输入如何引起正确时空转录响应。在这项研究中，我将探讨BMP信号传导的两种潜在机制。首先，新方法将允许我测定沿BMP梯度的途径活动的信号传导动力学。使用这些数据我将建立一个输入输出关系模型，以预测驱动BMP响应的机制转录动力学。其次，我将确定配体和受体对的不同组合是否播放 BMP梯度的规范和解释中的作用。我将研究如何动态和组合信号传导用于在胚胎中产生BMP信号反应的野生型模式。然后，使用在传播和塑造形态梯度的关键途径成分中的遗传扰动，我将测试动态和组合信号中的这些输入输出关系是否在不同的BMP上保持真实信号环境。这项研究的成功完成将为如何保守提供定量观点信号通路是由整个时间和空间的体内细胞解释的。