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 眼睛发育的标志是一个关键的胚胎过程，即腹侧视裂的闭合。故障发生在 视裂的适当闭合会导致缺损，这是严重视力障碍的一个原因。小胶质细胞， 驻留的中枢神经系统的天然免疫吞噬细胞，被发现在 视裂的闭合。小胶质细胞专门用于识别和消除凋亡细胞，以及 重要的是，细胞死亡与人和鼠的视裂闭合有关。然而，这是未知的， 小胶质细胞是否参与哺乳动物视裂的适当闭合。 为了解决这个问题，Aim1将确定小胶质细胞是否通过消除 凋亡性细胞。我将收集Bax-/-，MerTK-/-和PU.1-/-胚胎以及它们的产仔对照胚胎的组织，地址是 视裂闭合开始、视裂闭合期间和视裂闭合后。这些鼠标 LINES将允许我测试是否阻止发育中的细胞凋亡，阻止小胶质细胞识别 细胞凋亡或消除小胶质细胞完全改变了视裂的闭合。AIM2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Cx3CR1GFP/+收集组织 小鼠视裂闭合及视裂相关基因表达谱的测定 小胶质细胞与邻近视网膜腹侧小胶质细胞的原位杂交链式反应(HCR)。 虽然我在神经科学方面有很强的背景和实验室经验，但在 对于发展和疾病，我需要额外的培训才能发展成为一名独立的研究人员。我作为一名 研究生将在神经免疫学和学习贡献的技术和工具方面建立力量 从免疫学到神经发育。这个基础将使我为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，同时还为未来的拨款申请提供初步数据。在.之下 由该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， 并在实验选择、设计、分析和交流方面独立。这些技能将推动 我正在迈向独立研究人员的职业生涯。这项工作的完成将定义如何 小胶质细胞塑造视裂的闭合，并有助于视网膜和 视神经。这项建议旨在解决小胶质细胞在哺乳动物视裂闭合中的作用， 对涉及视裂闭合的通路的洞察，以及更广泛地揭示小胶质细胞如何调节 组织形态发生。