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 将人体器官再生为原来的器官仍然是我们想象的一部分。然而，在这方面， 蝾螈有再生大部分组织和器官的独特能力， 包括透镜。如果蝾螈的透镜丢失或受伤，它会从背部虹膜再生。 透镜再生具有重要的临床意义，也是研究晶状体再生的理想方法 组织再生许多离体技术，如组织学分析，只能 在特定时间从特定角度向我们展示再生过程的快照 点然而，透镜再生是一个涉及细胞、分子和细胞生物学的动态过程。 功能变化。我们已经能够，第一次，非侵入性地获得高质量的 光学相干断层扫描（OCT）在透镜再生过程中的体内图像 通过跟踪一只蝾螈超过40天。更有趣的是，OCT能够成像 脆弱的小带纤维在这个过程中第一次。这尚未使用 组织学/免疫组织学分析。此外，虹膜基质中的血管也是 清晰可见在这项提案中，我们将大大推进目前的成像技术， available.我们假设，通过整合高分辨率OCT和共焦 荧光激光扫描显微镜（CFLSM），结合使用蝾螈 在虹膜和血统追踪转基因蝾螈中缺乏色素，我们将能够在体内 图像的分子，细胞，和功能的变化发生的过程中， 透镜再生。为了实现这一目标，我们将定制一个多模态成像系统， 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，我们将揭示透镜再生的详细过程，包括透镜囊泡的形成， 透镜纤维的分化、悬韧带纤维的发育、虹膜血管的变化 以及使用突变体和转基因的组合的巨噬细胞的动态分布 蝾螈完成这些目标后，我们将建立新的全面成像 该平台将使研究人员能够在体内跟踪透镜再生的过程， 蝾螈没有中断和贡献的关键信息，可用于了解 白内障生物学、悬韧带病和透镜置换，其中完整的透镜囊缺失， 人类