Functional contribution of Metabolism in embryonic development

新陈代谢在胚胎发育中的功能贡献

基本信息

批准号：
10701781
负责人：
Mamiko Yajima
金额：
$ 23.93万
依托单位：
BROWN 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：
2022-09-12 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10701781
关键词：
Address Antisense Oligonucleotides Area Biological Biological Models Biology Biosensor Body Size Calcium Cell Differentiation process Cell Lineage Cell division Cell physiology Cells Cellular biology Chemicals Confocal Microscopy DNA biosynthesis Data Development Developmental Biology Embryo Embryonic Development Endomesoderm Event Fatty Acids Fertilization Genes Glucose Glycolysis Hour Image Mediating Metabolic Metabolic Pathway Metabolism Modeling Molecular Morphology Organism Outcome Oxidation-Reduction Oxidative Phosphorylation Oxygen Pathway interactions Pattern Phenotype Physiologic pulse Physiological Play Protein Biosynthesis Proteins Pyruvate Regulation Reporting Research Role Sea Urchins Signal Transduction Specific qualifier value Testing Time Visualization Warburg Effect Work aerobic glycolysis beta catenin biosynthetic product blastocyst blastomere structure cancer cell cell fate specification cell type cellular development embryo cell experimental study gastrulation gene regulatory network in vivo inhibitor membrane synthesis metabolomics protein biomarkers protein expression sensor temporal measurement tool

项目摘要

PROJECT SUMMARY Aerobic glycolysis was first identified in cancer cells (Warburg effect). This unusual metabolic process is considered to supply necessary biosynthetic products to sustain DNA, protein, and membrane synthesis in highly proliferative cells. However, the actual physiological significance of glycolysis remains largely unknown. Recent reports suggest that aerobic glycolysis may directly influence cellular function and development beyond matching a cell’s biosynthetic demands. Indeed, in our preliminary metabolomics results using the sea urchin embryo as a model system, dynamic metabolic regulation appears to be present throughout embryogenesis and further critical for a specific cell signaling event that occurs at the 16-cell stage of the embryo (5 hours post fertilization; 5hpf). This signaling event is called “micromere signaling” and known to drive endomesodermal specification in the entire embryo at two days post fertilization (2dpf). Based on these preliminary findings, we hypothesize that dynamic metabolic regulation serves as another layer of mechanism for cell specification and signaling during embryogenesis. To prove this hypothesis, in the proposed research, we will first visualize metabolic dynamics in real time and in vivo throughout embryogenesis, using GFP-tagged metabolic sensors. Imaging will be performed by 4D-confocal microscopy to maximize the spatial and temporal resolution of metabolic dynamics, which will be further subject to quantitative analysis for each cell lineage and for each developmental stage. Second, we will test the functional significance of each metabolic pathway (Glycolysis, Fatty Acid Synthesis, Oxidative Phosphorylation), especially in the event of micromere signaling at the 16-cell stage. Multiple inhibitors for each metabolic pathway will be applied for ~0.5 hour to the entire embryo or specifically to the micromeres at the 16-cell stage. The latter will be accomplished by constructing chimeric embryos in which the micromeres are replaced with the inhibitor-treated micromeres. The phenotypes will be then scored by analyzing temporal and spatial expression of polarity factors and fate determinants that drive the micromere- specific Gene Regulatory Network and is critical for endomesodermal specification, as well as overall morphology (e.g. successful gastrulation) in the resultant embryos. These experiments will identify the essentiality of each metabolic pathway to entire embryonic patterning. Overall, the proposed research will reveal the functional interplay of metabolic, gene and protein regulations essential for embryonic development, which is still understudied in the field of cell and developmental biology.

项目概要有氧糖酵解首先在癌细胞中被发现（瓦尔堡效应），这种不寻常的代谢过程是。被认为提供必要的生物合成产品来维持 DNA、蛋白质和膜的合成然而，糖酵解的实际生理意义在很大程度上仍然存在。最近的报告表明有氧糖酵解可能直接影响细胞功能和事实上，在我们的初步代谢组学中，发展超出了细胞生物合成的需求。使用海胆胚胎作为模型系统的结果，动态代谢调节似乎是存在于整个胚胎发生过程中，并且对于发生在胚胎发育过程中的特定细胞信号传导事件进一步至关重要。胚胎的 16 细胞阶段（受精后 5 小时；5hpf）此信号事件称为“微粒”。信号传导”，已知在两天后驱动整个胚胎的内中胚层规范受精 (2dpf) 基于这些初步发现，我们捕获了动态代谢。调节作为另一层机制，服务于细胞规范和信号传导为了证明这一假设，在拟议的研究中，我们将首先可视化代谢。使用 GFP 标记的代谢传感器，在整个胚胎发生过程中实时观察体内动态。成像将通过 4D 共焦显微镜进行，以最大限度地提高空间和时间代谢动力学的解析，将进一步对每个细胞进行定量分析其次，我们将测试每个发展阶段的功能意义。代谢途径（糖酵解、脂肪酸合成、氧化磷酸化），特别是在 16 细胞阶段的微米信号传导事件将针对每个代谢途径产生多种抑制剂。应用于整个胚胎或特别是 16 细胞阶段的微粒约 0.5 小时。后者将通过构建嵌合胚胎来实现，其中微团是然后用抑制剂处理的微粒替换。然后通过分析对表型进行评分。驱动微分子的极性因素和命运决定因素的时间和空间表达特定的基因调控网络，对于内中胚层规范以及这些实验将观察所得胚胎的整体形态（例如成功的原肠胚形成）。确定每个代谢途径对整个胚胎模式的重要性。拟议的研究将揭示代谢、基因和蛋白质调节之间的功能相互作用用于胚胎发育，细胞和发育生物学领域仍在对其进行研究。