Kim Dahyeon, Seo Jiseong, Jeon Taeck Joong
Department of Integrative Biological Sciences & BK21 FOUR Educational Research Group for Age-Associated Disorder Control Technology, Chosun University, Gwangju 61452, Republic of Korea.
The Basic Science Institute of Chosun University, Chosun University, Gwangju 61452, Republic of Korea.
J Microbiol Biotechnol. 2025 Apr 27;35:e2412066. doi: 10.4014/jmb.2412.12066.
Intracellular calcium (Ca) plays a vital role in numerous cellular processes, including signal transduction, cell motility, and development. Despite extensive research on intracellular calcium dynamics, the specific mechanisms by which extracellular calcium influences intracellular calcium levels remain unclear. In this study, we generated a -specific plasmid expressing the calcium biosensor GCaMP3 and visualized real-time intracellular calcium fluctuations in live cells. In response to external calcium, vegetative cells displayed a rapid and transient increase in intracellular calcium levels, peaking within 20 s, whereas aggregation-competent cells showed no such response. -null cells showed a fast and slight induction at approximately 5 s, with a slower and more efficient, extended high induction at 30 s. Meanwhile, -null cells showed a slightly delayed response, and a significantly lower proportion of -null cells responded to external calcium stimuli compared to wild-type cells. In the presence of W7, a calmodulin antagonist that inhibits calcium release from acidic stores, cells exhibited a marked reduction in the major calcium induction peak at 20 s post-stimulation. Our results suggest that there are at least two pathways to increase the intracellular calcium level in response to external calcium stimuli, and that intracellular acidic calcium stores partially contribute to the second major peak of calcium induction following external stimuli. It appears that G proteins, cGMP, and IplA are involved in calcium homeostasis upon external calcium stimulation, and play an important role in modulating the timing and amplitude of calcium responses.
细胞内钙(Ca)在众多细胞过程中发挥着至关重要的作用,包括信号转导、细胞运动和发育。尽管对细胞内钙动力学进行了广泛研究,但细胞外钙影响细胞内钙水平的具体机制仍不清楚。在本研究中,我们构建了一个表达钙生物传感器GCaMP3的特异性质粒,并可视化了活细胞中实时的细胞内钙波动。响应外部钙刺激,营养细胞显示细胞内钙水平迅速且短暂升高,在20秒内达到峰值,而具有聚集能力的细胞则无此反应。缺失 - 的细胞在大约5秒时显示出快速且轻微的诱导,在30秒时诱导速度较慢但更有效且持续时间更长。同时,缺失 - 的细胞显示出反应略有延迟,与野生型细胞相比,对外部钙刺激有反应的缺失 - 的细胞比例显著更低。在存在W7(一种抑制酸性储存库中钙释放的钙调蛋白拮抗剂)的情况下,细胞在刺激后20秒时主要钙诱导峰明显降低。我们的结果表明,至少有两条途径可响应外部钙刺激来提高细胞内钙水平,并且细胞内酸性钙储存部分促成了外部刺激后钙诱导的第二个主要峰值。看来G蛋白、cGMP和IplA在外部钙刺激时参与钙稳态,并在调节钙反应的时间和幅度方面发挥重要作用。