Danionella cerebrum as a transparent vertebrate adult model for studying immune-related biological processes and diseases

大脑丹尼奥菌作为透明脊椎动物成年模型，用于研究免疫相关的生物过程和疾病

• 批准号: 10665376
• 负责人: Pui Ying Lam
• 金额: $ 23.4万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665376
• 关键词: Acceleration; Adult; Affect; Aging; Animal Model; Animals; Behavioral; Biological Models; Biological Process; Biology; Brain; Calcium; Cell physiology; Cells; Central Nervous System; Cephalic; Cerebrum; Characteristics; Collection; Communities; Complex; DNA; Data; Data Set; Development; Disease; Disease Progression; Disease model; Dorsal; Endothelial Cells; Environment; Equipment; Event; Female; Future; Gene Transfer Techniques; Genetic Engineering; Goals; Heart; Heterogeneity; Image; Imaging Techniques; Immune; Immune response; Immune system; Immunological Models; Immunology; Inflammaging; Infrastructure; Injury; Institution; Kidney; Knowledge; Label; Larva; Learning; Length; Light; Liver; Macrophage; Malignant Neoplasms; Measures; Mechanics; Memory; Methods; Microglia; Microscopy; Modeling; Molecular; Mus; Names; Natural Immunity; Natural regeneration; Nature; Nerve Regeneration; Nervous System; Neurodegenerative Disorders; Optics; Organ; Physiological; Plasmids; Play; Procedures; Process; Reagent; Regenerative capacity; Regenerative research; Reporter; Research; Resolution; Role; Sex Differences; Study models; Systems Biology; Tail; Techniques; Testing; Therapeutic; Tissue imaging; Tissues; Transgenic Animals; Transgenic Organisms; Vascular System; Vertebrates; Visualization; Work; Zebrafish; adaptive immunity; addiction; age related; aged; awake; body cavity; body system; bone; cell type; comparative; cranium; experience; experimental study; genome editing; high resolution imaging; human disease; imaging modality; imaging study; immune cell infiltrate; improved; in vivo; in vivo Model; in vivo imaging; infancy; intravital imaging; male; model organism; mutant; neutrophil; novel; optogenetics; regenerative; response; response to injury; screening; sex; small molecule; teleost; tissue injury; tissue regeneration; tool; transcriptomics; two-photon; wound healing; young adult; Acceleration; Adult; Affect; Aging; Animal Model; Animals; Behavioral; Biological Models; Biological Process; Biology; Brain; Calcium; Cell physiology; Cells; Central Nervous System; Cephalic; Cerebrum; Characteristics; Collection; Communities; Complex; DNA; Data; Data Set; Development; Disease; Disease Progression; Disease model; Dorsal; Endothelial Cells; Environment; Equipment; Event; Female; Future; Gene Transfer Techniques; Genetic Engineering; Goals; Heart; Heterogeneity; Image; Imaging Techniques; Immune; Immune response; Immune system; Immunological Models; Immunology; Inflammaging; Infrastructure; Injury; Institution; Kidney; Knowledge; Label; Larva; Learning; Length; Light; Liver; Macrophage; Malignant Neoplasms; Measures; Mechanics; Memory; Methods; Microglia; Microscopy; Modeling; Molecular; Mus; Names; Natural Immunity; Natural regeneration; Nature; Nerve Regeneration; Nervous System; Neurodegenerative Disorders; Optics; Organ; Physiological; Plasmids; Play; Procedures; Process; Reagent; Regenerative capacity; Regenerative research; Reporter; Research; Resolution; Role; Sex Differences; Study models; Systems Biology; Tail; Techniques; Testing; Therapeutic; Tissue imaging; Tissues; Transgenic Animals; Transgenic Organisms; Vascular System; Vertebrates; Visualization; Work; Zebrafish; adaptive immunity; addiction; age related; aged; awake; body cavity; body system; bone; cell type; comparative; cranium; experience; experimental study; genome editing; high resolution imaging; human disease; imaging modality; imaging study; immune cell infiltrate; improved; in vivo; in vivo Model; in vivo imaging; infancy; intravital imaging; male; model organism; mutant; neutrophil; novel; optogenetics; regenerative; response; response to injury; screening; sex; small molecule; teleost; tissue injury; tissue regeneration; tool; transcriptomics; two-photon; wound healing; young adult

An optically accessible adult vertebrate model is needed to study dynamic cellular processes in vivo. We aim to develop and characterize Danionella cerebrum, a teleost related to zebrafish that remains transparent into adulthood, for studying immune-related biological processes and their impact on disease progression in multiple organs. A transparent adult model is particularly important for live imaging studies of immune related processes due to the dynamic and complex nature of immune cell heterogeneity and plasticity, the involvement of infiltrating immune cells during disease progression, as well as confounding factors related to aging. D. cerebrum, previously named as D. translucida, was first described by the Judkewitz group in 2018 for brain-related studies. This genetically tractable model is small, adults reach a length of approximately 12 mm in length, and is optical transparency even into adulthood. Established transgenesis and genome editing methods used in zebrafish have showed to be effective in D. cerebrum. These characteristics make D. cerebrum an attractive model for noninvasive in vivo visualization of dynamic and complex cellular events in a physiologically relevant setting. This study aims to build the infrastructure required to use D. cerebrum as a systems biology model organism for studying immune-related biological processes and their impact on disease progression. Our proof of principle experiments have successfully generated transgenic lines with fluorescently labelled innate immune cells such as neutrophils and macrophages, and the endothelial cells of the vasculature. We propose to expand our collection of transgenic D. cerebrum lines by fluorescently labeling multiple organs such as the brain, heart, liver, and kidney. In combination, these transgenic lines will allow us to visualize the complex interplay between immune cells and various organs throughout development, injury and disease. We will also develop a macrophage/microglia fluorescent activity reporter line which will allow us to observe and measure dynamic state changes displayed by immune cells in a physiologically relevant environment using non-invasive in vivo imaging. To facilitate our studies, we will develop methods and equipment for longitudinal live imaging of awake adult D. cerebrum. Immune cell functions play important roles in wound healing and regeneration. Zebrafish are known to have great regeneration capacity in different organs, however, it is not known if D. cerebrum can regenerate. Our preliminary work suggests that adult D. cerebrum can regenerate its tail fin but this regenerative capacity decreased in aged animal. Our proposed study will investigate if D. cerebrum regenerates its central nervous system during different developmental stages, its corresponding immune cell response and the transcriptomic changes that accompany these events. We will also perform interspecies comparative analysis using available data from zebrafish regeneration studies to explore key players in modulating regeneration capacity. This proposal describes strategies that will build the infrastructure required to use D. cerebrum for immunology related studies in vivo. The tools, techniques and knowledge built for the D. cerebrum immune model will provide an important springboard for carrying out future focused analyses on immune-related processes that include, but by no means are limited to, inflammaging, wound healing/regeneration, cancer, and neurodegenerative diseases.

需要一个可光学上的成年脊椎动物模型来研究体内动态细胞过程。我们旨在发展 并描述了与斑马鱼有关的Danionella Cerebrum，它是一个透明至成年的硬质的，用于研究 免疫相关的生物学过程及其对多个器官疾病进展的影响。透明的成人模型 由于免疫相关过程的实时成像研究尤其重要 免疫细胞的异质性和可塑性，疾病进展过程中浸润性细胞的参与 作为与衰老有关的混杂因素。 D.大脑，以前称为D. Cresscomida，首先是由 Judkewitz组于2018年进行与大脑有关的研究。这种遗传性拖延模型很小，成年人达到长度 长度约为12毫米，甚至是成年期的光学透明度。已建立的转基因和基因组 斑马鱼中使用的编辑方法已显示出在大脑D.脑中有效。这些特征使D.大脑 在生理上相关的动态和复杂细胞事件的非侵入性体内可视化的有吸引力模型 环境。这项研究旨在建立使用D.大脑作为系统生物学模型生物所需的基础设施 研究与免疫相关的生物学过程及其对疾病进展的影响。我们的原理实验证明 已经成功地产生了具有荧光标记的先天免疫细胞的转基因线，例如中性粒细胞和 巨噬细胞和脉管系统的内皮细胞。我们建议扩大我们的转基因D.大脑的收集 线条通过荧光标记多个器官，例如大脑，心脏，肝脏和肾脏。结合这些转基因 线条将使我们能够在整个发育过程中可视化免疫细胞与各种器官之间的复杂相互作用， 伤害和疾病。我们还将开发巨噬细胞/小胶质细胞荧光活性报告基因线，这将使我们得以 使用并测量免疫细胞在生理相关环境中显示的动态状态变化 非侵入性体内成像。为了促进我们的学习，我们将开发纵向现场的方法和设备 清醒成人D.大脑的成像。免疫细胞功能在伤口愈合和再生中起重要作用。 斑马鱼在不同的器官中具有很高的再生能力，但是，尚不清楚大脑是否可以 再生。我们的初步工作表明，成年大脑可以再生其尾鳍，但这种再生能力 老年动物减少。我们提出的研究将调查D.大脑在期间是否再生其中枢神经系统 不同的发育阶段，相应的免疫细胞反应以及随附的转录组变化 这些事件。我们还将使用斑马鱼再生的可用数据进行种间比较分析 研究探索主要参与者调节再生能力的研究。该建议描述了将建立的策略 在体内使用D.大脑进行相关研究所需的基础设施。工具，技术和知识 为D.大脑免疫模型建造将为对未来的专注分析提供重要的跳板 免疫相关的过程包括但绝不包括炎症，伤口愈合/再生，癌症， 和神经退行性疾病。