Microglial contributions to the development of the mammalian optic stalk

小胶质细胞对哺乳动物视柄发育的贡献

• 批准号: 10679913
• 负责人: Nathaniel Ghena
• 金额: $ 3.73万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10679913
• 关键词: Address; Age; Apolipoprotein E; Apoptosis; Apoptotic; Applications Grants; Automobile Driving; CSF1R gene; Cd68; Cell Death; Cells; Central Nervous System; Chick; Coloboma; Communication; Congenital Disorders; Cues; Data; Dependence; Development; Developmental Process; Disease; Embryo; Embryonic Development; Embryonic Eye; Event; Eye; Eye Development; Failure; Fellowship; Foundations; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Goals; Gold; HTATIP2 gene; Homeostasis; Human; Immune; Immunology; In Situ Hybridization; Influentials; Label; Mammals; Manuscripts; Mediating; Micro Array Data; Microarray Analysis; Microglia; Morphogenesis; Mus; Mutant Strains Mice; Myeloid Cells; Neuroimmune; Neurosciences; Optic Nerve; Optic vesicle; Optics; Participant; Pathway interactions; Phagocytes; Phagocytosis; Play; Process; Proteolysis; Reaction; Regulation; Research; Research Personnel; Retina; Role; Scientist; Shapes; Signal Transduction; Technical Expertise; Techniques; Testing; Tissues; Training; Visual impairment; Work; career; design; graduate student; innate immune pathways; insight; laboratory experience; lipid metabolism; neurodevelopment; neuroimmunology; novel; optic stalk; postnatal; receptor; recruit; response; selective expression; skills; stem; tool

Project Summary Ocular development is marked by a key embryonic process, the closure of the ventral optic fissure. Failure in the proper closure of the optic fissure results in coloboma, a cause of significant visual impairment. Microglia, the resident innate immune phagocyte of the central nervous system, are found to populate the retina prior to the closure of the optic fissure. Microglia are specialized to recognize and eliminate apoptotic cells, and critically, cell death is associated with optic fissure closure in mouse and human. It is unknown, however, whether microglia participate in the proper closure of the optic fissure in mammals. To address this, Aim1 will determine whether microglia facilitate optic fissure closure through elimination of apoptotic cells. I will collect tissue from Bax-/-, MerTK-/-, and Pu.1-/- embryos, and their littermate controls, at onset of optic fissure closure, during optic fissure closure, and following optic fissure closure. These mouse lines will allow me to test whether blocking developmental apoptosis, blocking microglial recognition of apoptotic cells, or eliminating microglia completely alters optic fissure closure. Aim2 will determine the expression profile of optic fissure-associated microglia during closure. I will collect tissue from Cx3CR1GFP/+ mice during optic fissure closure and determine the gene expression profile of optic fissure-associated microglia versus adjacent ventral retina microglia using in situ hybridization chain reaction (HCR). While I have a strong background and laboratory experience in neuroscience, both in the context of development and disease, I require additional training to develop as an independent researcher. My goals as a graduate student are to build strength in neuroimmunology and techniques and tools to study contributions of immunology to neurodevelopment. This foundation will prepare me to pursue my long-term career goal of becoming a research scientist. I anticipate that my preliminary data plus the aims described here will produce at least one scientific manuscript while also supplying preliminary data for future grant applications. Under the training plan provided by this fellowship, will refine technical skills, deepen my expertise in neuroimmunology, and become independent in experimental choice, design, analysis, and communication. These skills will propel me on my trajectory towards a career as an independent researcher. Completion of this work will define how microglia shape the closure of the optic fissure and contribute to the embryonic development of the retina and optic nerve. This proposal aims to address the role of microglia in mammalian optic fissure closure, yielding insights into pathways involved in optic fissure closure, and more broadly, revealing how microglia regulate tissue morphogenesis.

项目摘要 眼的发育以一个关键的胚胎过程为标志，即腹侧视裂的闭合。衰竭 视神经裂的适当闭合导致缺损，这是严重视力损害的原因。小胶质细胞， 中枢神经系统固有的先天免疫吞噬细胞，被发现在视网膜上， 视裂的闭合。小胶质细胞专门识别和消除凋亡细胞， 重要的是，细胞死亡与小鼠和人类的视裂闭合有关。然而，目前尚不清楚， 小胶质细胞是否参与哺乳动物视裂的正常闭合。 为了解决这个问题，Aim 1将确定小胶质细胞是否通过消除视神经裂来促进视神经裂闭合。 凋亡细胞本人将在以下时间点采集Bax-/-、MerTK-/-和Pu.1-/-胚胎及其同窝对照的组织： 视裂闭合开始、视裂闭合期间和视裂闭合后。这些小鼠 线将允许我测试是否阻断发育性凋亡，阻断小胶质细胞识别， 凋亡细胞或完全消除小胶质细胞改变视裂闭合。AIM 2将决定 视裂相关小胶质细胞在闭合过程中的表达谱。我将从Cx 3CR 1GFP/+中采集组织 小鼠视裂闭合过程中，并确定视裂相关的基因表达谱 小胶质细胞与相邻的腹侧视网膜小胶质细胞使用原位杂交链反应（HCR）。 虽然我在神经科学方面有很强的背景和实验室经验， 发展和疾病，我需要额外的培训，以发展为一个独立的研究人员。我的目标是 研究生将在神经免疫学和技术和工具方面建立实力，以研究 免疫学到神经发育这个基础将为我追求我的长期职业目标做好准备， 成为一名研究科学家我预计我的初步数据加上这里描述的目标将产生 至少一份科学手稿，同时也为未来的资助申请提供初步数据。下 这个奖学金提供的培训计划，将完善技术技能，加深我在神经免疫学方面的专业知识， 在实验选择、设计、分析和交流方面独立自主。这些技能将推动 我正朝着独立研究员的方向前进。完成这项工作将确定如何 小胶质细胞形成视裂的闭合，并有助于视网膜的胚胎发育， 视神经该提案旨在解决小胶质细胞在哺乳动物视裂闭合中的作用，得出 深入了解参与视裂闭合的途径，更广泛地说，揭示了小胶质细胞如何调节 组织形态发生