Nita Iulia I, Hershfinkel Michal, Kantor Chase, Rutter Guy A, Lewis Eli C, Sekler Israel
Department of Physiology and Cell Biology and.
Section of Cell Biology, Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, Faculty of Medicine, Imperial College London, London, UK.
FASEB J. 2014 Aug;28(8):3301-12. doi: 10.1096/fj.13-248161. Epub 2014 Apr 9.
Communication between the plasma membrane and mitochondria is essential for initiating the Ca(2+) and metabolic signals required for secretion in β cells. Although voltage-dependent Na(+) channels are abundantly expressed in β cells and activated by glucose, their role in communicating with mitochondria is unresolved. Here, we combined fluorescent Na(+), Ca(2+), and ATP imaging, electrophysiological analysis with tetrodotoxin (TTX)-dependent block of the Na(+) channel, and molecular manipulation of mitochondrial Ca(2+) transporters to study the communication between Na(+) channels and mitochondria. We show that TTX inhibits glucose-dependent depolarization and blocks cytosolic Na(+) and Ca(2+) responses and their propagation into mitochondria. TTX-sensitive mitochondrial Ca(2+) influx was largely blocked by knockdown of the mitochondrial Ca(2+) uniporter (MCU) expression. Knockdown of the mitochondrial Na(+)/Ca(2+) exchanger (NCLX) and Na(+) dose response analysis demonstrated that NCLX mediates the mitochondrial Na(+) influx and is tuned to sense the TTX-sensitive cytosolic Na(+) responses. Finally, TTX blocked glucose-dependent mitochondrial Ca(2+) rise, mitochondrial metabolic activity, and ATP production. Our results show that communication of the Na(+) channels with mitochondria shape both global Ca(2+) and metabolism signals linked to insulin secretion in β cells.- Nita, I. I., Hershfinkel, M., Kantor, C., Rutter, G. A., Lewis, E. C., Sekler, I. Pancreatic β-cell Na(+) channels control global Ca(2+) signaling and oxidative metabolism by inducing Na(+) and Ca(2+) responses that are propagated into mitochondria.
质膜与线粒体之间的通讯对于启动β细胞分泌所需的Ca(2+)和代谢信号至关重要。尽管电压依赖性Na(+)通道在β细胞中大量表达并被葡萄糖激活,但其在与线粒体通讯中的作用仍未明确。在此,我们结合荧光Na(+)、Ca(2+)和ATP成像、用河豚毒素(TTX)依赖性阻断Na(+)通道进行电生理分析以及对线粒体Ca(2+)转运体进行分子操作,以研究Na(+)通道与线粒体之间的通讯。我们发现TTX抑制葡萄糖依赖性去极化,并阻断胞质Na(+)和Ca(2+)反应及其向线粒体的传播。线粒体Ca(2+)单向转运体(MCU)表达的敲低在很大程度上阻断了TTX敏感的线粒体Ca(2+)内流。线粒体Na(+)/Ca(2+)交换体(NCLX)的敲低和Na(+)剂量反应分析表明,NCLX介导线粒体Na(+)内流,并经调节以感知TTX敏感的胞质Na(+)反应。最后,TTX阻断了葡萄糖依赖性线粒体Ca(2+)升高、线粒体代谢活性和ATP产生。我们的结果表明,Na(+)通道与线粒体的通讯塑造了与β细胞胰岛素分泌相关的全局Ca(2+)和代谢信号。- 尼塔,I.I.,赫什芬克尔,M.,坎托,C.,鲁特,G.A.,刘易斯,E.C.,塞克勒,I. 胰腺β细胞Na(+)通道通过诱导传播到线粒体的Na(+)和Ca(2+)反应来控制全局Ca(2+)信号传导和氧化代谢