Developmental Progression Driving Gastrulation of the Drosophila Early Embryo

驱动果蝇早期胚胎原肠胚形成的发育进程

基本信息

批准号：
9330885
负责人：
Angelike Stathopoulos
金额：
$ 57.82万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-11 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9330885
关键词：
Adhesives Animals Automobile Driving Biological Assay Biological Models Cell Nucleus Cell Signaling Process Cells Chromatin Data Development Developmental Process Dorsal Drosophila genus Embryo Event Exhibits Fibroblast Growth Factor Gene Expression Gene Expression Profiling Gene Targeting Genes Goals Heparan Sulfate Proteoglycan In Situ Hybridization Mesoderm Cell Methods Molecular Molecular Biology Molecular Conformation Molecular Profiling Morphogenesis Movement Neoplasm Metastasis Pattern Property Regulation Regulator Genes Regulatory Element Research Resolution Role Signal Pathway Signal Transduction System Time cell motility cohesion design gastrulation gene conservation gene therapy imaging approach imaging modality improved in vivo in vivo imaging insight interest mathematical model mutant new technology programs public health relevance spatiotemporal transcription factor

项目摘要

DESCRIPTION (provided by applicant): The overlying goal of the proposed research program is to understand the molecular mechanisms that control gastrulation using the Drosophila embryo as a model system. In particular, we are interested in investigating how dorsal-ventral (DV) patterning and cell signaling processes are temporally regulated in the early embryo; in deciphering the cis-regulatory mechanisms controlling spatiotemporal gene expression; and studying how collective cell movements are orchestrated. These are inter-related questions that also are relevant for the development of all animals, and as such these studies have the potential to provide far-reaching insights. To assay progression of developmental events, we develop and employ novel technologies for making temporally relevant observations using live in vivo imaging, computation including mathematical modeling, and molecular biology. We have focused on the design and implementation of new imaging approaches that allow us to acquire fine-scale spatiotemporal data of developmental processes, to capture transcription factor dynamics as well as cell movements. Here we propose three research directions to provide further insight into the system of genes driving Drosophila gastrulation. Project 1 involves expansion of DV patterning network with a focus on the regulation of temporal expression. In the early embryo, we have found that transcription factors acting along the DV axis exhibit dynamic changes in levels. Expression profiles for putative target genes at multiple time-points spanning the early development will be obtained at single embryo resolution in wildtype versus mutant embryos to provide insight into how timing of expression is regulated by the gene network. Spatial expression of a subset of genes will be further investigated using in situ hybridization, and live imaging methods will be used to monitor gene expression in real-time. Project 2 will investigate the mechanism and role of coordinate cis-regulatory action using methods to analyze chromatin conformation in vivo within each nucleus of the embryo as well as assays to compare levels and timing of gene expression supported by co-acting cis-regulatory elements. Project 3 will investigate the function and regulation of FGF signaling in migrating cells. We hypothesize that FGF signaling modulates the adhesive properties of mesoderm cells at gastrulation to support their cohesive, organized movement. The role of heparan sulfate proteoglycan molecules and cleavage-state on FGF activity will also be investigated. The overlying goal of the proposed research program is to understand the molecular mechanisms controlling morphogenesis of the embryo through study of a network of genes that controls patterning, signaling pathway activation, and, ultimately, cell movements in the early Drosophila embryo. The conservation of gene regulatory mechanisms across all animals promises that these studies will have far reaching implications. In particular, a better understanding of cis-regulatory mechanisms, in general, has many benefits including improved, targeted gene therapy; while understanding how cell migration is controlled will provide insights toward the regulation of cell metastasis.

描述（由申请人提供）：拟议研究计划的首要目标是了解使用果蝇胚胎作为模型系统控制原肠胚形成的分子机制，特别是，我们有兴趣研究背腹（DV）模式如何形成和控制。细胞信号传导过程在早期胚胎中受到时间调节；破译控制时空基因表达的顺式调节机制；以及研究集体细胞运动是如何协调的。也与所有动物的发育相关，因此这些研究有可能提供深远的见解，为了分析发育事件的进展，我们开发并采用新技术，利用活体成像、计算进行时间相关的观察。我们专注于新成像方法的设计和实施，这些方法使我们能够获取发育过程的精细时空数据，捕获转录因子动态以及细胞运动。提供进一步深入了解基因驱动系统的方向果蝇原肠胚形成涉及 DV 模式网络的扩展，重点是早期胚胎中的时间表达调节，我们发现沿 DV 轴作用的转录因子在水平上表现出动态变化。将在野生型与突变型胚胎的单个胚胎分辨率下获得跨越早期发育的多个时间点，以深入了解基因网络如何调节表达时间。将使用原位进一步研究基因子集的空间表达。杂交和活体成像方法将用于实时监测基因表达，项目2将使用分析胚胎每个细胞核内染色质构象的方法以及分析方法来研究协调顺式调控作用的机制和作用。比较共同作用的顺式调控元件支持的基因表达水平和时间。项目 3 将研究 FGF 信号在迁移细胞中的功能和调节。硫酸乙酰肝素蛋白聚糖分子和裂解状态对 FGF 活性的作用也将得到研究，该研究计划的首要目标是通过研究来了解控制胚胎形态发生的分子机制。控制早期果蝇胚胎中的模式、信号通路激活以及最终细胞运动的基因网络。所有动物中基因调控机制的保守性预示着这些研究将产生深远的影响。特别是，更好地了解顺式调控机制通常有很多好处，包括改进的靶向基因治疗；同时了解细胞迁移的控制方式将为细胞转移的调控提供见解。