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

基本信息

批准号：
9120882
负责人：
Kristen M Kwan
金额：
$ 29.8万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9120882
关键词：
Accounting Actins Adhesions Animal Model Anterior Biological Models Cell Death Cell Polarity Cell Survival Cell physiology Cells Choroid Collagen Collagen Type IV Coloboma Complex Computational Technique Computing Methodologies Defect Development Ectoderm Embryo Event Extracellular Matrix Eye Eye Development Failure Four-dimensional Fresh Water Glycoproteins Goals Health Hereditary nephritis Human Image Image Analysis Imaging Techniques Irido-corneo-trabecular dysgenesis Laminin Lead Lens Placodes Life Microscopy Molecular Molecular Genetics Morphogenesis Movement Mutate Mutation Neural Retina Newborn Infant Nidogen Ocular Pathology Optic vesicle Optics Pathway interactions Play Positioning Attribute Process Retina Role Series Shapes Signal Transduction Structural defect Structure Structure of retinal pigment epithelium Testing Tissues Vertebrates Visual impairment Zebrafish base cell behavior cell motility constriction crosslink eye formation imaging modality in vivo innovation lens molecular dynamics mutant novel optic cup prospective quantitative imaging research study teleost fish

项目摘要

DESCRIPTION (provided by applicant): Developmental defects in eye structure commonly account for visual impairment in newborns. Proper eye structure is initially established during the process of optic cup morphogenesis, during which the optic vesicle transforms into the optic cup via a series of complex cell and tissue rearrangements, with neural retina and retinal pigmented epithelium surrounding the newly formed lens. Recent advances in live imaging have begun to reveal the cellular processes underlying optic cup morphogenesis, yet molecular control of these critical events still remains largely unknown. A compelling candidate to play a role in controlling optic cup morphogenesis is the extracellular matrix (ECM), a complex, glycoprotein-rich layer that can regulate cell survival, movement, polarity, and tissue tension. ECM components, including laminin and type IV collagens, surround eye structures throughout optic cup morphogenesis in all vertebrates examined to date. Mutations in certain ECM components can lead to specific ocular pathologies in humans and model organisms, such as coloboma (failure of choroid fissure development) or lens defects. This suggests distinct requirements for ECM components during specific events in optic cup morphogenesis, yet this has not been examined in a systematic way. Zebrafish presents an ideal model system to study this process: optical transparency and rapid development offer a unique opportunity to directly watch eye formation in vivo. Further, we previously developed 4-dimensional imaging and computational techniques to track and visualize cell movements and behaviors throughout optic cup morphogenesis. This puts us in a unique position to quantitatively determine specific morphogenetic defects arising when particular matrix components are disrupted. In this proposal, we will determine the roles of laminin, type IV collagens, and the laminin-collagen cross-linking molecule nidogen during optic cup morphogenesis. I hypothesize that sequential steps of optic cup morphogenesis harbor distinct requirements for matrix components and higher-order assembly, and that these differential requirements underlie specific ocular pathologies seen in humans and model organisms. Combining molecular genetics with innovative 4-dimensional live imaging and computational methods, we will test this hypothesis in the following specific aims: (1) determine the role of matrix components and supramolecular assembly in choroid fissure formation; (2) determine how matrix components control optic cup invagination, taking advantage of a novel mutant, in which retina-lens de-adhesion (and possibly collagen signaling) are impaired; (3) determine the role of matrix components in establishing proper lens shape and ectoderm separation. The experiments proposed here will define the cellular and molecular dynamics of extracellular matrix adhesion underlying critical steps in optic cup morphogenesis, and the specific cellular functions executed by matrix during each step.

描述（由应用程序提供）：眼睛结构中的发育缺陷通常解释新生儿的视觉障碍。最初在光学杯形态发生过程中建立了适当的眼睛结构，在此期间，光学蔬菜通过一系列复杂的细胞和组织重排转化为视杯，并在新形成的透镜周围具有神经素和视网膜猪的上皮。实时成像的最新进展已经开始揭示了视觉杯形态发生的基础细胞过程，但是对这些关键事件的分子控制仍然很大程度上尚不清楚。令人信服的候选者在控制视觉杯形态发生中发挥作用是细胞外基质（ECM），这是一种复杂的，富含糖蛋白的层，可以调节细胞存活，运动，极性和组织张力。 ECM成分，包括层粘连蛋白和IV型胶原蛋白，在迄今已检查的所有脊椎动物中，整个视觉杯形态发生的眼睛结构。某些ECM组件中的突变会导致人类和模型组织中的特定眼病理性，例如Coloboma（脉络膜裂痕发育失败）或晶状体缺陷。这表明在光学杯形态发生的特定事件中，对ECM组件的不同要求，但尚未以系统的方式进行检查。斑马鱼提出了研究此过程的理想模型系统：光学透明度和快速发展为直接观察体内眼睛形成的独特机会。此外，我们以前开发了四维成像和计算技术，以跟踪和可视化整个视频杯形态发生的细胞运动和行为。这使我们处于独特的位置，以定量确定特定的形态发生缺陷，当特定矩阵组件中断时会产生。在此提案中，我们将确定层粘连蛋白，IV型胶原蛋白和层粘连蛋白 - 胶原蛋白的交联分子nidogen在光学杯形态发生过程中的作用。我假设光学杯形态发生的顺序步骤构成了对基质组件和高阶组件的不同要求，并且这些差异要求是人类和模型生物体中所见的特定眼科病理。将分子遗传学与创新的4维实时成像和计算方法相结合，我们将在以下特定目的中检验这一假设：（1）确定基质成分和超分子组装在脉络裂隙形成中的作用；（2）确定矩阵组件如何利用一种新型突变体来控制视觉杯的内陷，其中视网膜镜头去粘附（以及可能的胶原信号传导）受损；（3）确定基质成分在建立适当的晶状体形状和外胚层分离中的作用。此处提出的实验将定义细胞外基质粘合剂的细胞和分子动力学的临界步骤杯形形态发生以及每个步骤中矩阵执行的特定细胞功能。