Modeling and analysis of BMP-mediated Dorsal/Ventral patterning in zebrafish embryos

斑马鱼胚胎 BMP 介导的背侧/腹侧模式的建模和分析

基本信息

批准号：
10411944
负责人：
David Michael Umulis
金额：
$ 34.48万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10411944
关键词：
3-Dimensional Back Bone Morphogenetic Proteins Cells Computer software Coupled Data Development Diffusion Dorsal Elements Embryo Evaluation Experimental Models Feedback Gastrula Gene Expression Gene Expression Profile Genes Genetic Genetic Models Goals Image Impact evaluation Ligands Measurement Measures Mediating Modeling Neoplasm Metastasis Organ Organogenesis Pathway interactions Pattern Pattern Formation Performance Play Regulation Role Shapes Signal Transduction Signaling Protein Structure System Testing Three-dimensional analysis Time To specify Transforming Growth Factor beta Vertebrates Work Zebrafish angiogenesis antagonist base blastocyst chordin cytokine design dynamic system experimental study extracellular gastrulation imaging genetics insight loss of function mutation mathematical model member model development morphogens mutant protein function public health relevance quantitative imaging response segmentation Image analysis spatiotemporal

项目摘要

Project summary Bone Morphogenetic Proteins (BMPs) function as morphogens during development to specify spatial organization of embryonic axes and organ structures. Numerous extracellular antagonists and co-factors work in concert to dynamically control BMP signaling distributions along the dorsal-ventral (DV) embryonic axis in vertebrates to induce space and time-dependent patterns of gene expression. As the embryo progresses from the blastula stage into the gastrula stage, zygotic feedback in response to BMP signaling begins and cells start to play an active role by simultaneously responding to BMPs and regulating the secretion of antagonists and other factors that shape the BMP gradient. However it is not well understood how the system dynamically regulates pattern formation. Studies of gastrula stage BMP pattern formation provide an ideal context to unravel how feedback regulation shapes the gradients of BMP signaling in a vertebrate system. We hypothesize that BMP-mediated feedback by admp, sizzled, tld, and bambi dynamically shape the gradient in gastrula embryos. Our objective is to delineate how coupled patterning and feedback shape gradients and discover mechanisms of BMP regulation in zebrafish gastrula. In Aim 1 we will quantify spatiotemporal BMP signaling and feedback target gene expression in wild-type and mutant zebrafish gastrula embryos. Furthermore, we will measure phospho-Smad 1/5 (PSmad) levels in toto in wild-type (wt) gastrula embryos, and develop our new wavelet-based segmentation and image analysis software WaveSeg (https://waveletseg.weebly.com) to quantify, segment, and register the data. This work will provide the first- ever quantitative data needed to identify feedback relationships that shape the PSmad gradient. Quantification of BMP patterning in gastrula stage embryos will provide insight into the role of feedback in gradient refinement in a rapidly changing spatial domain. In Aim 2 we will develop a 3-dimensional growing mesh finite element model of BMP pattern formation in gastrula stage embryos. Evaluation of the models against quantitative imaging data provides a rigorous basis to discern the mechanisms of gastrula embryo pattern formation and generate new tests to validate or refute the consistent mechanisms. The seamless integration and back-and- forth between imaging, genetics, and modeling will elucidate how the extracellular BMP pathway factors modulate the BMP signaling gradient. Lastly, in Aim 3 we will investigate the feedback loops in BMP signaling that have been proposed to provide embryonic scale invariance, robustness with respect to partial loss of network components, and increase the dynamic range of signal interpretation. Understanding these networks of BMP regulation will provide insight into BMP function in other contexts including organogenesis, cancer metastasis, angiogenesis and development.

项目摘要在发育过程中，骨形态发生蛋白（BMP）充当形态剂，以指定空间胚胎轴和器官结构的组织。许多细胞外拮抗剂和联合因素工作协同沿着背腹侧（DV）胚胎轴的动态控制BMP信号传导分布脊椎动物诱导基因表达的空间和时间依赖性模式。随着胚胎的发展囊泡阶段进入胃阶段，响应BMP信号传导的二合作反馈开始，细胞开始通过同时响应BMP并调节对抗者的分泌和塑造BMP梯度的其他因素。但是，它不太了解系统如何动态调节模式形成。胃阶段BMP模式形成的研究为解开反馈调节如何塑造脊椎动物系统中BMP信号的梯度。我们假设ADMP，Sizzled，TLD和Bambi介导的反馈，动态地塑造了梯度胃胚胎。我们的目标是描述如何耦合图案和反馈形状梯度和发现斑马鱼胃中BMP调节的机制。在AIM 1中，我们将量化时空BMP 信号传导和反馈靶基因表达在野生型和突变的斑马鱼胃胚胎中。此外，我们将在野生型（WT）胃胚胎中测量磷酸化1/5（PSMAD）水平，并开发我们新的基于小波的细分和图像分析软件waveseg （https://waveletseg.weebly.com）量化，细分和注册数据。这项工作将提供第一确定塑造PSMAD梯度的反馈关系所需的定量数据。定量 BMP在胃阶段胚胎中的图案将提供有关反馈在梯度改进中的作用的洞察力在快速变化的空间域中。在AIM 2中，我们将开发一个三维生长的网格有限元 BMP模式形成模型在胃阶段胚胎中。评估模型针对定量的评估成像数据提供了严格的基础，以辨别胃胚胎模式形成的机制和生成新的测试以验证或反驳一致的机制。无缝集成和来回成像，遗传学和建模之间的第四将阐明细胞外BMP途径如何因素调节BMP信号梯度。最后，在AIM 3中，我们将研究BMP信号中的反馈回路提议提供胚胎量表的不变性，鲁棒性有关网络组件，并增加信号解释的动态范围。了解这些网络 BMP调控的洞察力将在其他情况下洞悉BMP功能，包括器官发生，癌症转移，血管生成和发育。