Intracellular signaling during phagocytosis

吞噬过程中的细胞内信号传导

基本信息

批准号：
8319303
负责人：
Kevin Swier
金额：
$ 14.5万
依托单位：
CHICAGO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8319303
关键词：
Animal Model Bacteria Biological Assay Biological Models Biological Sciences Biology Biomedical Research Cell Culture Techniques Cell Differentiation process Cell Line Cell Separation Cell physiology Cells Cellular biology Chicago Communities Complex Data Defect Development Dictyostelium Dictyostelium discoideum Eating Educational Curriculum Educational process of instructing Educational workshop Endocytosis Eukaryotic Cell Extramural Activities Fluorescence Food Funding Future Genes Genetic Genetic Screening Goals Green Fluorescent Proteins Human Immune Immune response Immune system Institution Latex Bead Lectin Link Mainstreaming Mediating Mentors Metabolic Metabolism Methods Microbe Minority Minority-Serving Institution Organism Phagocytes Phagocytosis Phagosomes Physiologic pulse Physiological Pilot Projects Plasmids Population Principal Investigator Process Program Development Property Proteins Public Health Recruitment Activity Regulation Reporter Research Resource Sharing Screening procedure Signal Pathway Signal Transduction Sorting - Cell Movement Source Staging Students Time Universities Vertebrates career cell growth cell transformation cell type design detection of nutrient experience fluorescence activated cell sorter device innovation insight meetings microorganism mutant novel premature professor programs promoter red fluorescent protein response restriction enzyme undergraduate education

项目摘要

DESCRIPTION (provided by applicant): This application seeks funding for a pilot project that will allow the PI to chart a new direction in research. The PI is a productive Professor of Biology in the Department of Biological Sciences at Chicago State University, a minority-serving institution. The PI was recently introduced to the model organism Dictyostelium discoideum and has been using this organism to introduce students in classrooms and summer workshops to basic cell biology research. This experience has inspired the PI to establish a new research program that builds upon his experience in the cell biology of endocytosis and phagocytosis. The innovative screens of Dictyostelium mutants proposed here will immediately establish a new lab setting in which to mentor students and set a new research agenda for the PI. The PI aims to establish his presence in the Dictyostelium community, and gather preliminary results that will support proposals for funding from mainstream extramural sources. Phagocytosis, or cell-eating, is a complex process carried out by many eukaryotic cells, especially those of vertebrate immune systems. It is especially important to understand how phagocytic cells respond to the variety of microorganisms they encounter. Dictyostelium is a professional phagocyte and is the only model organism that allows forward genetic approaches to studying the fundamental cell biology of phagocytosis. Two novel fluorescent assays are proposed that will be used to screen Dictyostelium mutants for defects in signaling during phagocytosis. Dictyostelium cells will be mutagenized by restriction enzyme-mediated integration (REMI) of a plasmid expressing a selectable marker. In the first screen, Dictyostelium mutants will be pulsed with fluorescent bacteria and sorted for cells that cannot degrade the bacteria. For the second screen, a Dictyostelium cell line will be constructed that fluoresces upon induction of the prestarvation response. Dictyostelium mutants will then be screened for inability to inhibit this response in the presence of bacteria, their primary food source. These assays will identify novel genes involved in intracellular signaling during phagocytosis. Because phagocytosis in Dictyostelium is necessarily linked to decisions regarding cell growth and differentiation, we expect to establish Dictyostelium as a model system for investigating cellular mechanisms that integrate nutrient sensing, metabolic regulation and development.

描述（由申请人提供）：本申请为试点项目寻求资金，该项目将允许PI绘制研究的新方向。 PI是芝加哥州立大学生物科学系生物学的生产教授，这是一家少数族裔服务机构。 PI最近被引入模型生物体迪斯特尔迪斯特尔，并一直在使用该生物体向教室和夏季研讨会中的学生介绍基本的细胞生物学研究。这种经验激发了PI建立一个新的研究计划，该计划基于他在内吞和吞噬作用的细胞生物学方面的经验。此处提出的Dictyostelium突变体的创新屏幕将立即建立一个新的实验室环境，以指导学生并为PI设定新的研究议程。 PI的目的是建立他在Dictyostelium社区中的存在，并收集初步结果，以支持主流壁外来源资助的建议。吞噬作用或吞噬细胞是许多真核细胞，尤其是脊椎动物免疫系统的复杂过程。了解吞噬细胞如何应对它们遇到的微生物的种类尤为重要。 dictyostelium是一种专业的吞噬细胞，是唯一允许研究吞噬细胞增多基本细胞生物学的远期遗传方法的模型生物。提出了两种新型的荧光测定，该测定法将用于筛选dictyostelium突变体，以使吞噬作用期间的信号传导缺陷。柱状细胞将通过表达可选标记物的质粒的限制酶介导的整合（REMI）进行诱变。在第一个屏幕上，柱状骨突变体将用荧光细菌脉冲，并分类无法降解细菌的细胞。对于第二个屏幕，将构建柱状细胞系，该细胞系在诱导预脱位响应后会产生荧光。然后将筛选dictyostelium突变体，以免在细菌存在的主要食物来源的存在下抑制这种反应。这些测定将确定吞噬作用期间参与细胞内信号传导的新基因。由于dictyostelium中的吞噬作用必然与细胞生长和分化的决策有关，因此我们希望将dictyostelium建立为研究整合营养感应，代谢调节和发育的细胞机制的模型系统。