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

摘要 目前资助的研究2R15GM124670-02专注于发现钙离子的作用 铜绿假单胞菌与宿主细胞相互作用中的信号转导。这些研究包括建立 通过监测赤潮藻钙离子水平的变化，探讨钙离子动态平衡的信号模式。 铜绿假单胞菌与宿主细胞的相互作用，包括侵袭、细胞内存活和 逃走。使准确测量钙离子的时空变化成为可能 在铜绿假单胞菌细胞亚细胞间隙内的浓度，我们要求替换目前 过时的平板阅读器Biotek，配备了更新版本的仪器Cytation 5。新的 该仪器增强了测量能力和测量精度。它允许低噪音 基于单色仪的吸光度、可调荧光和发光的读数。 重要的是，它还将包括使用20X-60X物镜、高对比度亮场和 四种不同的荧光成像立方体。它允许通过4个分区进行严格的温度控制 温度控制系统，防止板盖结露。它提供双试剂 能够进行多模式检测的注射器。所要求的仪器将被放置在PI的实验室中 将可供PI实验室和Dr。 卢特。所有参与的学生都将接受培训，以使用该乐器及其附带的 软件由公司代表提供。预期的研究进展将显著 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的能力。 所获得的新知识将增进我们对这种重要的人类病原体的了解。 并通知开发更有效的方法来控制或预防其致命的 感染。

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.