Metabolic and Oxidative Stress Effects on the Spectroelectrochemical Behavior of Single Pseudomonas aeruginosa Cells.
Metabolic and Oxidative Stress Effects on the Spectroelectrochemical Behavior of Single Pseudomonas aeruginosa Cells.
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DOI：
10.1021/cbmi.3c00083
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发表时间：
2023-10-23
期刊：
影响因子：
--
通讯作者：
Bohn, Paul W
中科院分区：
文献类型：
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作者：
Cutri, Allison R;Shrout, Joshua D;Bohn, Paul W
铜绿假单胞菌是一种机会性人类病原体，能够在免疫功能低下的患者中引起多种疾病。为了更好地了解铜绿假单胞菌的行为和毒力并推进对抗感染的药物治疗，了解铜绿假单胞菌细胞如何在应激条件下生存，特别是环境应激源将是有益的。在这里，我们报告了一种测量单个铜绿假单胞菌细胞的电位依赖性荧光的策略，作为细胞对饥饿、饥饿和氧化应激的反应的哨兵。这是通过使用微孔电极阵列来实现的，该微孔电极阵列能够在明确的空间位置捕获大量孤立的、垂直定向的细胞，以便并行研究大型单细胞阵列。我们发现，促进饥饿或氧化应激的条件会产生荧光响应的明显变化，这可以通过荧光瞬变的发生率增加来证明，荧光瞬变是单个铜绿假单胞菌细胞表现出的三种典型光谱电化学行为之一。相反，与稳定期的健康细胞相比，更适度的营养限制对光谱电化学反应几乎没有影响。这些发现证明了微孔电极阵列在细胞间距、方向和化学环境受控的条件下研究单个微生物细胞行为的能力。在如此明确的条件下实现单细胞研究使得以前所未有的控制来研究基本应激反应成为可能。
Pseudomonas aeruginosa is an opportunistic human pathogen capable of causing a wide range of diseases in immunocompromised patients. In order to better understand P. aeruginosa behavior and virulence and to advance drug therapies to combat infection, it would be beneficial to understand how P. aeruginosa cells survive stressful conditions, especially environmental stressors. Here, we report on a strategy that measures potential-dependent fluorescence of individual P. aeruginosa cells, as a sentinel, for cellular response to starvation, hunger, and oxidative stress. This is accomplished using a micropore electrode array capable of trapping large numbers of isolated, vertically oriented cells at well-defined spatial positions in order to study large arrays of single cells in parallel. We find that conditions promoting either starvation or oxidative stress produce discernible changes in the fluorescence response, demonstrated by an increase in the prevalence of fluorescence transients, one of three canonical spectroelectrochemical behaviors exhibited by single P. aeruginosa cells. In contrast, more modest nutrient limitations have little to no effect on the spectroelectrochemical response when compared to healthy cells in the stationary phase. These findings demonstrate the capabilities of micropore electrode arrays for studying the behavior of single microbial cells under conditions where the intercellular spacing, orientation, and chemical environment of the cells are controlled. Realizing single-cell studies under such well-defined conditions makes it possible to study fundamental stress responses with unprecedented control.