Calcium signaling controls Pseudomonas aeruginosa invasion and adaptation to the host intracellular environment

钙信号控制铜绿假单胞菌入侵和适应宿主细胞内环境

• 批准号: 10685112
• 负责人: Marianna Patrauchan
• 金额: $ 9.95万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10685112
• 关键词: Bacteria; Behavior; Calcium; Calcium Signaling; Cell physiology; Cells; Clinical; Computer software; Detection; Development; Environment; Eukaryota; Event; Fluorescence; Funding; Homeostasis; Human; Image; Infection; Knowledge; Measurement; Molecular; Monitor; Noise; Pattern; Physical condensation; Process; Pseudomonas aeruginosa; Reader; Reading; Reagent; Research; Role; Signal Transduction; Students; Subcellular Spaces; System; Temperature; Training; Update; base; human pathogen; improved; instrument; luminescence; pathogenic bacteria; prevent; temporal measurement

ABSTRACT The currently funded research 2R15GM124670-02 is focused on discovering the role of Ca2+ signaling in the interactions of P. aeruginosa with host cells. These studies include establishing the signaling patterns of Ca2+ homeostasis by monitoring the changes of Ca2+ levels in P. aeruginosa during interactions with host cells, including invasion, intracellular survival, and escape. To enable the accurate measurements of the temporal and spatial changes of Ca2+ concentration within subcellular spaces of P. aeruginosa cells, we request to replace the currently obsolete plate reader Biotek with the updated version of the instrument, Cytation 5. The new instrument offers enhanced capabilities and accuracy of measurements. It allows low-noise reading of monochromator-based absorbance, tunable fluorescence, and luminescence. Importantly, it will also include imaging using 20X – 60X objectives, high contrast brightfield and four different imaging cubes for fluorescence. It allows tight temperature control with a 4-zone temperature control system and prevents condensation on plate lids. It offers dual reagent injectors able for multimode detection. The requested instrument will be placed in the PI's lab and will be available for the use by all the students in the PI's lab and the collaborating lab of Dr. Lutter. All the participating students will be trained to use the instrument and the accompanying software by the company representative. The anticipated research advances will significantly enhance our ability to study calcium signaling in P. aeruginosa interacting with its eukaryotic host. The obtained new knowledge will improve our understanding of this important human pathogen and inform the development of more efficient means for controlling or preventing its deadly infections.

抽象的 当前资助的研究2R15GM124670-02专注于发现CA2+的作用 铜绿假单胞菌与宿主细胞相互作用的信号传导。这些研究包括建立 通过监测P中Ca2+水平的变化，Ca2+稳态的信号传导模式。 在与宿主细胞相互作用的过程中，铜绿菌，包括侵袭，细胞内存活和 逃脱。为了实现CA2+的临时和空间变化的准确测量 铜绿假单胞菌细胞的亚细胞空间内的浓度，我们要求更换当前 过时的板读取器Biotek具有仪器的更新版本，Cytation 5。 仪器提供了增强的功能和测量的准确性。它允许低噪声 读取基于单色剂的吸收，可调荧光和发光。 重要的是，它还将包括使用20倍 - 60倍目标成像，高对比度Brightfield和 四个不同的成像立方体用于荧光。它允许使用4区的温度控制 温度控制系统并防止板盖上的凝结。它提供双试剂 喷油器可以用于多模检测。请求的仪器将放置在PI的实验室中， PI实验室中的所有学生和Dr. Dr.的合作实验室都可以使用。 丽特。所有参与的学生将接受培训以使用该工具和参与 公司代表软件。预期的研究进展将显着 增强我们研究铜绿假单胞菌与其真核宿主相互作用的钙信号传导的能力。 获得的新知识将改善我们对这种重要人类病原体的理解 并告知开发更有效的手段来控制或防止其致命 感染。