Pinton P, Pozzan T, Rizzuto R
Department of Biomedical Sciences and CNR Centre for the Study of Biomembranes, University of Padova, Via Colombo 3, 35121 Padova, Italy.
EMBO J. 1998 Sep 15;17(18):5298-308. doi: 10.1093/emboj/17.18.5298.
In the past few years, intracellular organelles, such as the endoplasmic reticulum, the nucleus and the mitochondria, have emerged as key determinants in the generation and transduction of Ca2+ signals of high spatio-temporal complexity. Little is known about the Golgi apparatus, despite the fact that Ca2+ within its lumen controls essential processes, such as protein processing and sorting. We report the direct monitoring of the [Ca2+] in the Golgi lumen ([Ca2+]Golgi) of living HeLa cells, using a specifically targeted Ca2+-sensitive photoprotein. With this probe, we show that, in resting cells, [Ca2+]Golgi is approximately 0.3 mM and that Ca2+ accumulation by the Golgi has properties distinct from those of the endoplasmic reticulum (as inferred by the sensitivity to specific inhibitors). Upon stimulation with histamine, an agonist coupled to the generation of inositol 1,4,5-trisphosphate (IP3), a large, rapid decrease in [Ca2+]Golgi is observed. The Golgi apparatus can thus be regarded as a bona fide IP3-sensitive intracellular Ca2+ store, a notion with major implications for the control of organelle function, as well as for the generation of local cytosolic Ca2+ signals.
在过去几年中,细胞内细胞器,如内质网、细胞核和线粒体,已成为高时空复杂性Ca2+信号产生和转导的关键决定因素。尽管高尔基体腔内的Ca2+控制着蛋白质加工和分选等重要过程,但人们对高尔基体的了解却很少。我们报告了使用一种特异性靶向的Ca2+敏感光蛋白对活HeLa细胞高尔基体腔内的[Ca2+]([Ca2+]Golgi)进行直接监测。利用该探针,我们发现,在静息细胞中,[Ca2+]Golgi约为0.3 mM,并且高尔基体对Ca2+的积累具有与内质网不同的特性(通过对特定抑制剂的敏感性推断)。在用组胺刺激后,组胺是一种与肌醇1,4,5-三磷酸(IP3)生成相关的激动剂,观察到[Ca2+]Golgi大幅快速下降。因此,高尔基体可被视为一个真正的IP3敏感的细胞内Ca2+储存库,这一概念对细胞器功能的控制以及局部胞质Ca2+信号的产生具有重要意义。