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: 10043483
• 负责人: Katia Del Rio-Tsonis
• 金额: $ 20.79万
• 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 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10043483
• 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 将人体器官再生为原始器官仍然是我们想象的一部分。然而， 蝾螈具有独特的能力，可以再生它们的大部分组织和器官，甚至 成人期，包括镜头。如果纽特晶状体丢失或受伤，它会从虹膜背部再生。 晶状体再生具有重要的临床意义，但也是一种理想的研究过程。 一般情况下，组织再生。许多体外技术，如组织分析，只能 向我们展示在特定时间从特定角度看再生过程的快照 指向。然而，晶状体再生是一个动态的过程，涉及细胞、分子和 功能变化。我们第一次能够以非侵入性的方式获得高质量的 光学相干断层扫描(OCT)在晶状体再生过程中的活体图像 通过在40天的时间里追踪一只蝾螈。更有趣的是，OCT能够想象 在这一过程中，首次发现了脆弱的带状纤维。这还没有被记录在使用 组织学/免疫组织学分析。此外，虹膜基质中的血管也 清晰可见。在这份提案中，我们将大幅推进目前的成像技术 可用。我们假设通过集成高分辨率OCT和共焦 荧光激光扫描显微镜(CFLSM)结合Newts的使用 在缺乏色素的虹膜和谱系追踪的转基因蝾螈中，我们将能够在体内 想象一下在这个过程中发生的分子、细胞和功能的变化 晶状体再生。为了实现这一点，我们将定制一个多模成像系统， 高分辨率、足够的成像深度和功能成像能力。在三个目标中 提出，我们将揭示晶状体再生的详细过程，包括晶状体囊泡的形成， 晶状体纤维分化、索状纤维发育、虹膜血管改变 利用突变和转基因相结合的方法研究巨噬细胞的动态分布 这是一只纽带。在完成这些目标之后，我们将建立一个新的综合成像 平台，这将使研究人员能够在体内跟踪单个 没有中断的Newt，并提供可以用来理解的关键信息 无完整晶状体囊的白内障生物学、晶状体透明带病和晶状体置换术 人类