Hajnóczky G, Hager R, Thomas A P
Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
J Biol Chem. 1999 May 14;274(20):14157-62. doi: 10.1074/jbc.274.20.14157.
The concerted action of inositol 1,4,5-trisphosphate (IP3) and Ca2+ on the IP3 receptor Ca2+ release channel (IP3R) is a fundamental step in the generation of cytosolic Ca2+ oscillations and waves, which underlie Ca2+ signaling in many cells. Mitochondria appear in close association with regions of endoplasmic reticulum (ER) enriched in IP3R and are particularly responsive to IP3-induced increases of cytosolic Ca2+ ([Ca2+]c). To determine whether feedback regulation of the IP3R by released Ca2+ is modulated by mitochondrial Ca2+ uptake, the interactions between ER and mitochondrial Ca2+ pools were examined by fluorescence imaging of compartmentalized Ca2+ indicators in permeabilized hepatocytes. IP3 decreased luminal ER Ca2+ ([Ca2+]ER), and this was paralleled by an increase in mitochondrial matrix Ca2+ ([Ca2+]m) and activation of Ca2+-sensitive mitochondrial metabolism. Remarkably, the decrease in [Ca2+]ER evoked by submaximal IP3 was enhanced when mitochondrial Ca2+ uptake was blocked with ruthenium red or uncoupler. Moreover, subcellular regions that were relatively deficient in mitochondria demonstrated greater sensitivity to IP3 than regions of the cell with a high density of mitochondria. These data demonstrate that Ca2+ uptake by the mitochondria suppresses the local positive feedback effects of Ca2+ on the IP3R, giving rise to subcellular heterogeneity in IP3 sensitivity and IP3R excitability. Thus, mitochondria can play an important role in setting the threshold for activation and establishing the subcellular pattern of IP3-dependent [Ca2+]c signaling.
肌醇1,4,5 - 三磷酸(IP3)和Ca2+对IP3受体Ca2+释放通道(IP3R)的协同作用是细胞质Ca2+振荡和波动产生过程中的一个基本步骤,而细胞质Ca2+振荡和波动是许多细胞中Ca2+信号传导的基础。线粒体似乎与富含IP3R的内质网(ER)区域紧密相连,并且对IP3诱导的细胞质Ca2+([Ca2+]c)增加特别敏感。为了确定释放的Ca2+对IP3R的反馈调节是否受线粒体Ca2+摄取的调节,通过对通透化肝细胞中分隔的Ca2+指示剂进行荧光成像,研究了内质网和线粒体Ca2+池之间的相互作用。IP3降低了内质网腔Ca2+([Ca2+]ER),与此同时线粒体基质Ca2+([Ca2+]m)增加以及Ca2+敏感的线粒体代谢被激活。值得注意的是,当用钌红或解偶联剂阻断线粒体Ca2+摄取时,亚最大浓度的IP3引起的[Ca2+]ER降低会增强。此外,线粒体相对较少的亚细胞区域对IP3的敏感性高于线粒体密度高的细胞区域。这些数据表明,线粒体对Ca2+的摄取抑制了Ca2+对IP3R的局部正反馈作用,从而导致IP3敏感性和IP3R兴奋性的亚细胞异质性。因此,线粒体在设定激活阈值和建立IP3依赖性[Ca2+]c信号传导的亚细胞模式方面可以发挥重要作用。