In vivo imaging of newt lens regeneration: Novel molecular, cellular and functional insights

蝾螈晶状体再生的体内成像：新颖的分子、细胞和功能见解

• 批准号: 10250409
• 负责人: Katia Del Rio-Tsonis
• 金额: $ 17.52万
• 依托单位: MIAMI UNIVERSITY OXFORD
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10250409
• 关键词: Adult; Animal Model; Animals; Biology; Blood Vessels; Cataract; Custom; Degenerative Disorder; Development; Dorsal; Electron Microscopy; Epithelial Cells; Eye; Fiber; Fluorescence; Functional Imaging; Genetic Models; Genome; Goals; Histologic; Human; Image; Imagination; Imaging technology; Immunofluorescence Immunologic; In Situ Hybridization; Injury; Interruption; Iris; Knowledge; Laser Scanning Microscopy; Lens Fiber; Life Cycle Stages; Mission; Molecular; Monophenol Monooxygenase; Morphology; Multimodal Imaging; Natural regeneration; Newts; Notophthalmus viridescens; Optical Coherence Tomography; Organ; Oxygen; Pigment Epithelium; Pigments; Pleurodeles; Process; Public Health; Research; Research Personnel; Resolution; Technology; Testing; Time; Tissues; Transgenic Organisms; United States National Institutes of Health; Vesicle; Work; anterior chamber; cellular imaging; clinically significant; density; eye chamber; imaging capabilities; imaging modality; imaging platform; imaging system; in vivo; in vivo imaging; injured; insight; lens; lens capsule; lens regeneration; macrophage; metabolic rate; molecular imaging; mutant; novel; tissue regeneration; tool

Project Summary Regenerating a human organ as its original one remains a part of our imagination. However, newts have unique capabilities of regenerating most of their tissues and organs, even into adulthood, including the lens. If the newt lens is lost or injured, it regenerates from the dorsal iris. Lens regeneration has important clinical significance, but it is also an ideal process for studying tissue regeneration in general. Many ex-vivo technologies, such as histological analysis, can only show us a snapshot of the regeneration process from a specific point of view at a specific time point. However, lens regeneration is a dynamic process involving cellular, molecular and functional changes. We have been able to, for the first time, non-invasively acquire high-quality in vivo images during the process of lens regeneration with optical coherence tomography (OCT) by tracking a single newt for over a period of 40 days. More interestingly, OCT was able to image the fragile zonular fibers for the first time during this process. This has not been documented using histological/immunohistological analysis. In addition, the blood vessels in the iris stroma are also clearly visible. In this proposal, we will significantly advance the imaging technology currently available. We hypothesize that by integrating high-resolution OCT and confocal fluorescence laser scanning microscopy (CFLSM), in combination with the use of newts lacking pigments in the iris and lineage tracing transgenic newts, we will be able to in vivo image the molecular, cellular, and functional changes taking place during the process of lens regeneration. To achieve this, we will custom-build a multimodality imaging system with high resolution, sufficient imaging depth, and functional imaging capabilities. In the three aims proposed, we will reveal the detail process of lens regeneration, including lens vesicle formation, lens fiber differentiation, the development of the zonular fibers, the changes in iris vasculature and the dynamic distribution of macrophages using a combination of mutant and transgenic newts. After completing these aims, we will have established a new comprehensive imaging platform, that will allow researchers to in vivo track the process of lens regeneration in a single newt without interruptions and contribute critical information that can be used to understand cataract biology, zonulopathies and lens replacement where intact lens capsules are absent in humans

项目摘要 再生作为原始器官仍然是我们想象力的一部分。然而， Newt具有独特的能力，可以再生其大多数组织和器官，甚至进入 成年，包括镜头。如果Newt镜头丢失或受伤，它将从背虹膜中再生。 镜头再生具有重要的临床意义，但它也是研究的理想过程 组织再生一般。许多前体技术，例如组织学分析，只能 从特定的角度向我们展示再生过程的快照 观点。然而，镜头再生是一个动态过程，涉及细胞，分子和 功能变化。我们第一次能够非侵入性获得高质量 光学相干断层扫描（OCT）的晶状体再生过程中的体内图像（OCT） 通过跟踪单个NEWT 40天的时间。更有趣的是，OCT能够形象 在此过程中，第一次脆弱的Zonular纤维。尚未使用 组织学/免疫组织学分析。另外，虹膜基质中的血管也是 清晰可见。在此提案中，我们将大大推进目前的成像技术 可用的。我们通过整合高分辨率OCT和共同点来假设这一点 荧光激光扫描显微镜（CFLSM），结合使用NEWTS 在虹膜中缺少颜料和谱系追踪转基因new，我们将能够体内 图像在过程中发生的分子，细胞和功能变化 镜头再生。为了实现这一目标，我们将使用 高分辨率，足够的成像深度和功能成像功能。在三个目标中 提出的，我们将揭示晶状体再生的细节过程，包括晶状体形成， 透镜纤维分化，缘纤维的发育，虹膜脉管系统的变化 以及使用突变体和转基因组合的巨噬细胞的动态分布 纽特。完成这些目标后，我们将建立一个新的综合成像 平台，这将使研究人员能够体内跟踪一个单一的镜头再生过程 Newt没有中断并贡献可用于理解的关键信息 白内障生物学，划界和透镜替代品，在完整的镜头胶囊中不存在 人类