Stepwise Coordination of Eye Morphogenesis by Extracellular Matrix

细胞外基质对眼睛形态发生的逐步协调

• 批准号: 10583547
• 负责人: Kristen M Kwan
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10583547
• 关键词: Automobile Driving; Basement membrane; Biological Models; Bruch' s basal membrane structure; Cell Death; Cell Size; Cell Survival; Cell Transplantation; Cell physiology; Cells; Chondroitin Sulfate A; Chondroitin Sulfate Proteoglycan; Collaborations; Coloboma; Complex; Computational Technique; Computer Analysis; Computing Methodologies; Data; Defect; Deposition; Development; Ectoderm; Embryo; Environment; Epithelium; Event; Exposure to; Extracellular Matrix; Eye; Eye Development; Focal Adhesions; Four-dimensional; Fresh Water; Gene Expression Profiling; Glycoproteins; Goals; Human; Image; Imaging Techniques; Impairment; In Vitro; Laminin; Methods; Microscopy; Modeling; Molecular; Molecular Genetics; Morphogenesis; Morphology; Movement; Mutation; Neural Crest; Neural Crest Cell; Neural Retina; Newborn Infant; Nidogen; Ocular Pathology; Optic vesicle; Optics; Organoids; Pathway interactions; Physiological; Positioning Attribute; Process; Regenerative Medicine; Retina; Role; Series; Shapes; Signal Transduction; Structural defect; Structure; Structure of retinal pigment epithelium; Tenascin; Testing; Tissue Engineering; Tissue Model; Tissues; Transplantation; Visual impairment; Visualization; Work; Zebrafish; cell behavior; cell motility; cell type; experimental study; extracellular; eye formation; imaging modality; in vivo; innovation; lens; membrane assembly; model organism; molecular dynamics; optic cup; progenitor; programs; protein crosslink; response; single cell sequencing; spatiotemporal; teleost fish; timeline; versican; zebrafish development

Project Summary Developmental defects in eye structure commonly account for visual impairment in newborns. Proper eye structure is initially established via the process of optic cup morphogenesis, during which a series of complex cell and tissue rearrangements transforms the optic vesicle into the optic cup, with neural retina and retinal pigmented epithelium (RPE) enwrapping the newly formed lens. With advances in imaging and computational analysis, work from our lab and others has begun to reveal the cellular events underlying optic cup morphogenesis, however, molecular control of these processes still remains poorly understood. A compelling candidate to play a role in controlling optic cup morphogenesis is the extracellular matrix (ECM), a complex, glycoprotein-rich layer that regulates cell survival, movement, signaling, and polarity. Mutations in certain ECM components can lead to ocular pathologies, such as coloboma, suggesting specific requirements during optic cup morphogenesis. Understanding extrinsic control of morphogenesis also has implications for organoid approaches and strategies. Our previous data indicate that a core molecule, laminin, elicits diverse cellular responses in different eye regions. We also found that separate eye domains are exposed to distinct ECM microenvironments, some of which are assembled via tissue-tissue interactions: specifically, neural crest is required to build basement membrane around the RPE. These data suggest that unique ECM microenvironments may be a crucial driver of regional eye morphogenetic events. Zebrafish provide an ideal model system to study this process: optical transparency and rapid development offer a unique opportunity to directly observe and molecularly dissect eye formation in vivo. We previously developed 4-dimensional imaging and computational techniques to track and visualize cell movements throughout optic cup morphogenesis, and recently, methods for automated quantitative analysis of retinal cell size, shape and orientation. This puts us in a unique position to analyze specific morphogenetic defects arising when particular matrix components are disrupted. In this proposal, we will dissect the region-specific roles of ECM factors, including nidogens, tenascin-C, mmp2, and versican, during eye morphogenesis. We hypothesize that dynamic, region-specific ECM microenvironments elicit unique developmental and morphogenetic responses from distinct eye progenitor domains to drive optic cup morphogenesis. Combining molecular genetics with innovative 4-dimensional live imaging and computational methods, we will test this hypothesis in the following specific aims: (1) determine how ECM microenvironment controls retina morphogenesis and organization; (2) determine how ECM modulatory factors control RPE morphogenesis; and (3) determine functional requirements for tissue contributions to specific ECM microenvironments. The experiments proposed will define the spatiotemporal dynamics of ECM activity and distinct cellular functions executed by region-specific ECM factors during crucial steps of eye formation.

项目摘要 眼睛结构中的发育缺陷通常解释新生儿的视觉障碍。适当的眼睛 结构最初是通过视杯形态发生的，在此期间建立的 细胞和组织重排将视线囊泡转化为视杯，并带有神经视网膜和视网膜 色素上皮（RPE）包装新形成的镜头。 随着成像和计算分析方面的进步，我们实验室和其他人的工作已经开始揭示 然而，光学杯形态发生的基础事件，但是，这些过程的分子控制仍然存在 理解不佳。令人信服的候选人在控制视觉杯形态发生中发挥作用是 细胞外基质（ECM）是一种复杂的，富含糖蛋白的层，可调节细胞的存活，运动，信号传导， 和极性。某些ECM成分中的突变会导致眼科病理，例如Coloboma， 提出视觉杯形态发生过程中的特定要求。了解外部控制 形态发生也对器官方法和策略具有影响。我们以前的数据表明 核心分子，层粘连蛋白会引起不同眼部区域的各种细胞反应。我们还发现那是分开的 眼域暴露于不同的ECM微环境，其中一些环境通过组织组织组装 相互作用：具体来说，在RPE周围建造基底膜需要神经冠。这些数据 表明独特的ECM微环境可能是区域眼形形态发生事件的关键驱动力。 斑马鱼提供了研究此过程的理想模型系统：光学透明度和快速发展 提供独特的机会，直接观察并分子在体内剖析眼睛形成。我们以前 开发了四维成像和计算技术，以跟踪和可视化细胞运动 在整个视频杯形态发生，以及最近的视网膜细胞自动定量分析方法 大小，形状和方向。这使我们处于独特的位置，可以分析发生的特定形态发生缺陷 当特定矩阵组件被破坏时。在此提案中，我们将剖析特定区域的角色 ECM因子在眼形发生过程中，包括NIDogens，Tenascin-C，MMP2和Versican。 我们假设那种动态的，特定区域的ECM微环境引起了独特的发展和 来自不同眼祖域的形态发生反应以驱动视杯形态发生。结合 具有创新的4维实时成像和计算方法的分子遗传学，我们将测试这一点 假设以下特定目的：（1）确定ECM微环境如何控制视网膜 形态发生和组织； （2）确定ECM调节因子如何控制RPE形态发生；和 （3）确定组织贡献对特定ECM微环境的功能要求。这 提出的实验将定义ECM活性的时空动力学和不同的细胞功能 在眼睛形成的关键步骤中由区域特异性ECM因子执行。