Quantitative image analysis to determine the function of selected microglia-expressed genes in retinal development and regeneration

项目总结 这份爱达荷州INBRE行政副刊将在INBRE-发展部之间建立合作 研究项目研究员S龙和IDEA共同资助的R01首席研究员D·米切尔。Dr。 龙是一位计算机科学家，在机器学习和计算机辅助图像分析方面拥有专业知识。Dr。 米切尔是一位细胞生物学家/免疫学家，在视网膜发育/再生方面拥有专业知识。该项目的标题是， ‘定量图像分析以确定视网膜中选定的小胶质细胞表达基因的功能 发展和再生“。具体目标有三个： 具体目标1.在两名调查员之间开展合作，以加强两个项目和 增加本科生研究机会。 特定目标2：确定视网膜对选定的小胶质细胞表达基因的需求 发展和再生。 具体目标3：开发图像分析管道以支持斑马鱼视网膜分析。 小胶质细胞是驻留在中枢神经系统中的吞噬细胞，吞噬和降解病原体， 凋亡的细胞和碎片。此外，这些细胞可能参与视网膜退行性疾病、损伤、 动态平衡、发育、再生和/或与穆勒胶质细胞的串扰。米切尔实验室做到了 斑马鱼再生视网膜分离的小胶质细胞/巨噬细胞种群的转录组分析。从… 这是五个小胶质细胞表达的感兴趣基因的入围名单。斑马鱼突变系现在是，或者将会 BE，为每个基因建立。实时共聚焦成像将在野生型、杂合型和 纯合斑马鱼品系记录视网膜发育或再生过程中的小胶质细胞动力学。《长河》 实验室将开发一条图像分析管道，以自动解释来自 这些实验。这些分析将量化小胶质细胞的数量和形态、细胞死亡、Müler胶质细胞 子代细胞的增殖、肥大、迁移和神经前体标志物的表达。这个 Long实验室开发的机器学习技术将缓解瓶颈和潜在的偏见 在分析来自Mitchell实验室的大型图像数据集并允许快速测试小胶质细胞基因时 功能。强大的初步数据和调查人员的专业知识表明，该团队将完成拟议的 工作。Long-Mitchell的合作将通过以下方式为学生提供连续的研究机会 提供更广泛的学生研究体验。龙博士拥有专业知识和环境来推动 学生进入数据科学，这是现代生物学研究的必要补充。本科生将 在以本科为主的刘易斯-克拉克州立学院的长期实验室中得到支持， 以及研究密集型爱达荷州大学的米切尔实验室。

PROJECT SUMMARY This Idaho INBRE Administrative Supplement will establish a collaboration between an INBRE-Developmental Research Program Investigator, S Long, and an IDeA co-funded R01 principle investigator, D Mitchell. Dr. Long is a computer scientist with expertise in machine learning and computer-assisted image analyses. Dr. Mitchell is a cell biologist/immunologist with expertise in retinal development/regeneration. The project is titled, ‘Quantitative image analysis to determine the function of selected microglia-expressed genes in retinal development and regeneration’. There are three Specific Aims: Specific Aim 1. Develop a collaboration between the two investigators to enhance both projects and increase undergraduate student research opportunities. Specific Aim 2: Determine the requirement for selected microglia-expressed genes in retinal development and regeneration. Specific Aim 3: Develop an image analysis pipeline to support zebrafish retinal analyses. Microglia are the resident phagocytes in the central nervous system and engulf and degrade pathogens, apoptotic cells, and debris. In addition, these cells may be involved in retinal degenerative disease, injury, homeostasis, development, regeneration, and/or crosstalk with Müller glia. The Mitchell laboratory did transcriptome analysis of microglia/macrophage populations isolated from regenerating zebrafish retinas. From this, a shortlist of five microglia-expressed genes of interest are identified. Zebrafish mutant lines are, or will be, established for each gene. Real-time live confocal imaging will be done on wild-type, heterozygous, and homozygous zebrafish lines to record microglial dynamics in developing or regenerating retinas. The Long laboratory will develop an image analysis pipeline to automate the interpretation of the large datasets from these experiments. The analyses will quantify microglia numbers and morphology, cell death, Müller glial proliferation, hypertrophy, migration of daughter cells, and expression of neural progenitor markers. The machine learning techniques developed by the Long laboratory will relieve the bottleneck and potential biases in analysis of large image datasets from the Mitchell laboratory and allow rapid testing of microglia gene function. Strong preliminary data and investigator expertise indicate that this team will complete the proposed work. The Long-Mitchell collaboration will provide a continuum of research opportunities for students by delivering broadened student research experiences. Dr. Long has the expertise and the environment to propel students into Data Science, a needed complement for modern biological research. Undergraduate students will be supported in both the Long laboratory at the primarily undergraduate institution, Lewis-Clark State College, and the Mitchell laboratory at the research-intensive, University of Idaho.