Department of Molecular, Cellular, and Developmental Biology, University of Colorado Boulder, Boulder, CO 80303.
BioFrontiers Institute, University of Colorado Boulder, Boulder, CO 80303.
Proc Natl Acad Sci U S A. 2017 Aug 29;114(35):9445-9450. doi: 10.1073/pnas.1703084114. Epub 2017 Aug 14.
Electrically excitable cells harness voltage-coupled calcium influx to transmit intracellular signals, typically studied in neurons and cardiomyocytes. Despite intense study in higher organisms, investigations of voltage and calcium signaling in bacteria have lagged due to their small size and a lack of sensitive tools. Only recently were bacteria shown to modulate their membrane potential on the timescale of seconds, and little is known about the downstream effects from this modulation. In this paper, we report on the effects of electrophysiology in individual bacteria. A genetically encoded calcium sensor expressed in revealed calcium transients in single cells. A fusion sensor that simultaneously reports voltage and calcium indicated that calcium influx is induced by voltage depolarizations, similar to metazoan action potentials. Cytoplasmic calcium levels and transients increased upon mechanical stimulation with a hydrogel, and single cells altered protein concentrations dependent on the mechanical environment. Blocking voltage and calcium flux altered mechanically induced changes in protein concentration, while inducing calcium flux reproduced these changes. Thus, voltage and calcium relay a bacterial sense of touch and alter cellular lifestyle. Although the calcium effectors remain unknown, these data open a host of new questions about , including the identity of the underlying molecular players, as well as other signals conveyed by voltage and calcium. These data also provide evidence that dynamic voltage and calcium exists as a signaling modality in the oldest domain of life, and therefore studying electrophysiology beyond canonical electrically excitable cells could yield exciting new findings.
可兴奋细胞利用电压门控钙内流来传递细胞内信号,这在神经元和心肌细胞中得到了广泛研究。尽管在高等生物中进行了深入研究,但由于细菌体积小且缺乏敏感工具,对细菌电压和钙信号的研究一直滞后。直到最近,人们才发现细菌能够在秒级的时间尺度上调节其膜电位,但对于这种调节的下游效应知之甚少。在本文中,我们报告了单个细菌的电生理学效应。在 中表达的一种遗传编码钙传感器揭示了单个细胞中的钙瞬变。同时报告电压和钙的融合传感器表明,钙内流是由电压去极化诱导的,类似于后生动物动作电位。用水凝胶进行机械刺激会增加细胞质钙水平和瞬变,而单个细胞会根据机械环境改变蛋白质浓度。阻断电压和钙流会改变机械诱导的蛋白质浓度变化,而诱导钙流则会重现这些变化。因此,电压和钙传递细菌的触觉,并改变细胞的生活方式。尽管钙效应物仍不清楚,但这些数据为 开辟了一系列新的问题,包括潜在的分子参与者的身份,以及电压和钙传递的其他信号。这些数据还提供了证据,表明动态电压和钙作为一种信号传递方式存在于生命最古老的领域,因此研究超越经典可兴奋细胞的电生理学可能会产生令人兴奋的新发现。