Collaborative Research: Characterization of a Bioluminescence Reporter System for Mechanical Stress

合作研究：机械应力生物发光报告系统的表征

• 批准号: 1205930
• 负责人: Michael Latz
• 金额: $ 27.65万
• 依托单位: University of California-San Diego Scripps Inst of Oceanography
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1205930&HistoricalAwards=false
• 关键词: Collaborative; Research; Characterization; Bioluminescence; Reporter

In this project, the PIs will investigate the relationship between mechanical properties of the cell and physiological response, using bioluminescence as a fast reporter of mechanosensitivity. Specific questions to be addressed are: (1) What are the mechanical properties of the cell? (2) How is mechanosensitivity related to the mechanical properties of the cell? and (3) What is the role of the cytoskeleton in determining the mechanical properties of cells and their physiological sensitivity? These issues will be studied in three species of dinoflagellates using atomic force microscopy to measure the elasticity of living cells and apply quantified mechanical stress to cells, microfluidics to subject cells to known levels of fluid shear stress, and laser confocal microscopy coupled with fluorescent probes to visualize the cytoskeleton. Pharmacological treatments that disrupt the cytoskeleton will be examined for their effect on cell elasticity and mechanosensitivity. This collaborative project applies advanced methods of physics, material science, and microfluidic technology to characterize the mechanical properties of the cell and the relationship to physiological sensitivity. The goal of the project is to enhance the value of dinoflagellate bioluminescence as a reporter of mechanical stress for applications in both the biological and physical sciences. This project will broaden the participation of underrepresented groups and advances discovery and understanding by providing internship opportunities to these undergraduate students through UCSD internship programs, the opportunity for a Ph.D. graduate student in the physical sciences to work in an interdisciplinary setting, and for two undergraduate students to participate as REU interns. The project advances discovery and understanding by contributing to the professional development of middle and high school science teachers by developing course materials through the Scripps Classroom Connection Summer Institute in partnership with COSEE CA, and providing demonstration kits of dinoflagellate bioluminescence to their students. Project results will be broadly disseminated through established scientific venues as well as education and public outreach activities through the Birch Aquarium at Scripps.

在该项目中，PI将使用生物发光作为机械敏感性的快速记者，研究细胞的机械性能与生理反应之间的关系。要解决的具体问题是：（1）细胞的机械性能是什么？ （2）机械敏感与细胞的机械性能有何关系？ （3）细胞骨架在确定细胞的机械性能及其生理敏感性中的作用是什么？这些问题将使用原子力显微镜在三种鞭毛藻中进行研究，以测量活细胞的弹性，并将机械应力施加到细胞中，将微流体施加到受试者细胞中，以对已知水平的流体剪切应力和激光共焦显微镜与荧光探针相连，以使细胞球骨骼可视化。破坏细胞骨架的药理学治疗对细胞弹性和机械敏感性的影响将检查。该协作项目应用了物理，材料科学和微流体技术的先进方法来表征细胞的机械性能以及与生理灵敏度的关系。该项目的目的是增强鞭毛藻生物发光的价值，作为在生物科学和物理科学中应用机械应力的记者。该项目将通过UCSD实习计划向这些本科生提供实习机会，从而扩大代表性不足的团体的参与，并进步发现和理解。物理科学的研究生将在跨学科的环境中工作，并让两名本科生作为REU实习生参加。该项目通过与Cosee CA合作开发课程材料来为中学和高中科学教师的专业发展做出贡献，从而提高发现和理解，并为学生提供恐龙生物发光的示范套件。项目结果将通过既定的科学场所以及通过Scripps的桦木水族馆进行的教育和公共外展活动进行广泛传播。