Plasma membrane large-conductance calcium-activated potassium (BK) channels are important players in various physiological processes, including those mediated by epithelia. Like other cell types, human bronchial epithelial (HBE) cells also express BK in the inner mitochondrial membrane (mitoBK). The genetic relationships between these mitochondrial and plasma membrane channels and the precise role of mitoBK in epithelium physiology are still unclear. Here, we tested the hypothesis that the mitoBK channel is encoded by the same gene as the plasma membrane BK channel in HBE cells. We also examined the impact of channel loss on the basic function of HBE cells, which is to create a tight barrier. For this purpose, we used CRISPR/Cas9 technology in 16HBE14o- cells to disrupt the gene, which encodes the α-subunit responsible for forming the pore of the plasma membrane BK channel. Electrophysiological experiments demonstrated that the disruption of the gene resulted in the loss of BK-type channels in the plasma membrane and mitochondria. We have also shown that HBE ΔαBK cells exhibited a significant decrease in transepithelial electrical resistance which indicates a loss of tightness of the barrier created by these cells. We have also observed a decrease in mitochondrial respiration, which indicates a significant impairment of these organelles.

中文翻译：

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BKCa 钾通道的缺乏对人类支气管上皮的生理学具有重要意义

质膜大电导钙激活钾 (BK) 通道在各种生理过程（包括上皮介导的生理过程）中发挥着重要作用。与其他细胞类型一样，人支气管上皮 (HBE) 细胞也在线粒体内膜 (mitoBK) 中表达 BK。这些线粒体和质膜通道之间的遗传关系以及 mitoBK 在上皮生理学中的确切作用仍不清楚。在这里，我们测试了这样的假设：mitoBK 通道与 HBE 细胞中的质膜 BK 通道由相同的基因编码。我们还研究了通道丢失对 HBE 细胞基本功能（即创建紧密屏障）的影响。为此，我们在 16HBE14o- 细胞中使用 CRISPR/Cas9 技术来破坏编码负责形成质膜 BK 通道孔的 α 亚基的基因。电生理学实验表明，该基因的破坏导致质膜和线粒体中 BK 型通道的丧失。我们还表明，HBE ΔαBK 细胞的跨上皮电阻显着降低，这表明这些细胞产生的屏障的紧密性丧失。我们还观察到线粒体呼吸减少，这表明这些细胞器受到显着损害。