Csordás G, Thomas A P, Hajnóczky G
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
EMBO J. 1999 Jan 4;18(1):96-108. doi: 10.1093/emboj/18.1.96.
Transmission of cytosolic [Ca2+] ([Ca2+]c) oscillations into the mitochondrial matrix is thought to be supported by local calcium control between IP3 receptor Ca2+ channels (IP3R) and mitochondria, but study of the coupling mechanisms has been difficult. We established a permeabilized cell model in which the Ca2+ coupling between endoplasmic reticulum (ER) and mitochondria is retained, and mitochondrial [Ca2+] ([Ca2+]m) can be monitored by fluorescence imaging. We demonstrate that maximal activation of mitochondrial Ca2+ uptake is evoked by IP3-induced perimitochondrial [Ca2+] elevations, which appear to reach values >20-fold higher than the global increases of [Ca2+]c. Incremental doses of IP3 elicited [Ca2+]m elevations that followed the quantal pattern of Ca2+ mobilization, even at the level of individual mitochondria. In contrast, gradual increases of IP3 evoked relatively small [Ca2+]m responses despite eliciting similar [Ca2+]c increases. We conclude that each mitochondrial Ca2+ uptake site faces multiple IP3R, a concurrent activation of which is required for optimal activation of mitochondrial Ca2+ uptake. This architecture explains why calcium oscillations evoked by synchronized periodic activation of IP3R are particularly effective in establishing dynamic control over mitochondrial metabolism. Furthermore, our data reveal fundamental functional similarities between ER-mitochondrial Ca2+ coupling and synaptic transmission.
胞质[Ca2+]([Ca2+]c)振荡向线粒体基质的传递被认为是由IP3受体Ca2+通道(IP3R)与线粒体之间的局部钙调控所支持的,但对其偶联机制的研究一直很困难。我们建立了一种通透细胞模型,其中内质网(ER)与线粒体之间的Ca2+偶联得以保留,并且线粒体[Ca2+]([Ca2+]m)可以通过荧光成像进行监测。我们证明,IP3诱导的线粒体外周[Ca2+]升高可引发线粒体Ca2+摄取的最大激活,其升高幅度似乎比[Ca2+]c的整体升高高出20倍以上。即使在单个线粒体水平,递增剂量的IP3也能引发遵循Ca2+动员量子模式的[Ca2+]m升高。相比之下,尽管引发了相似的[Ca2+]c升高,但IP3的逐渐增加引发的[Ca2+]m反应相对较小。我们得出结论,每个线粒体Ca2+摄取位点面对多个IP3R,同时激活这些位点是线粒体Ca2+摄取最佳激活所必需的。这种结构解释了为什么IP3R同步周期性激活引发的钙振荡在建立对线粒体代谢的动态控制方面特别有效。此外,我们的数据揭示了内质网-线粒体Ca2+偶联与突触传递之间的基本功能相似性。