Stable interactions between mitochondria and endoplasmic reticulum allow rapid accumulation of calcium in a subpopulation of mitochondria
Stable interactions between mitochondria and endoplasmic reticulum allow rapid accumulation of calcium in a subpopulation of mitochondria
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DOI：
10.1074/jbc.m302301200
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发表时间：
2003-10-03
影响因子：
4.8
通讯作者：
Pozzan, T
中科院分区：
文献类型：
--
作者：
Filippin, L;Magalhaes, PJ;Pozzan, T
为了更好地了解线粒体网络在塑造活细胞中钙信号方面的功能作用，我们利用了最新的基于绿色荧光蛋白的最新基因工程传感器(“Cameleons”、“Camgaroos”和“Pericam”)和经典的钙敏感光蛋白aequorin，它们都针对线粒体基质。详细分析了基于绿色荧光蛋白的探针在亚细胞定位、光敏性和钙亲和力等方面的性质。结论是，比率骨膜是目前单细胞研究中最可靠的线粒体钙探针，尽管该探针也存在问题。在单细胞中用比率计量型Pericam获得的结果，与用水飞蓟素在群体水平上获得的结果相结合，提供了强有力的证据，证明线粒体靠近钙释放通道(从而负责在受体激活时线粒体对钙的快速吸收)在时间上是高度稳定的，表明线粒体和内质网之间存在特异性的相互作用。
To better understand the functional role of the mitochondrial network in shaping the Ca2+ signals in living cells, we took advantage both of the newest genetically engineered green fluorescent protein-based Ca2+ sensors ("Cameleons," "Camgaroos," and "Pericams") and of the classical Ca2+-sensitive photoprotein aequorin, all targeted to the mitochondrial matrix. The properties of the green fluorescent protein-based probes in terms of subcellular localization, photosensitivity, and Ca2+ affinity have been analyzed in detail. It is concluded that the ratiometric pericam is, at present, the most reliable mitochondrial Ca2+ probe for single cell studies, although this probe too is not devoid of problems. The results obtained with ratiometric pericam in single cells, combined with those obtained at the population level with aequorin, provide strong evidence demonstrating that the close vicinity of mitochondria to the Ca2+ release channels (and thus responsible for the fast uptake of Ca2+ by mitochondria upon receptor activation) are highly stable in time, suggesting the existence of specific interactions between mitochondria and the endoplasmic reticulum.