Store-operated Ca2+ influx and subplasmalemmal mitochondria.
ABSTRACT Calcium depletion of the endoplasmic reticulum (ER) induces oligomerisation, puncta formation and translocation of the ER Ca(2+) sensor proteins, STIM1 and -2 into plasma membrane (PM)-adjacent regions of the ER, where they activate the Orai1, -2 or -3 proteins present in the opposing PM. These proteins form ion channels through which store-operated Ca(2+) influx (SOC) occurs. Calcium ions exert negative feed-back on SOC. Here we examined whether subplasmalemmal mitochondria, which reduce this feed-back by Ca(2+) uptake, are located within or out of the high-Ca(2+) microdomains (HCMDs) formed between the ER and plasmalemmal Orai1 channels. For this purpose, COS-7 cells were cotransfected with Orai1, STIM1 labelled with YFP or mRFP and the mitochondrially targeted Ca(2+) sensitive fluorescent protein inverse-Pericam. Depletion of ER Ca(2+) with ATP+thapsigargin (in Ca(2+)-free medium) induced the appearance of STIM1 puncta in the < or =100 nm wide subplasmalemmal space, as examined with TIRF. Mitochondria were located either in the gaps between STIM1-tagged puncta or in remote, STIM1-free regions. After addition of Ca(2+) mitochondrial Ca(2+) concentration increased irrespective of the mitochondrion-STIM1 distance. These observations indicate that mitochondria are exposed to Ca(2+) diffused laterally from the HCMDs formed between the PM and the subplasmalemmal ER.
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ABSTRACT: Cross-talk between organelles and plasma membrane Ca(2+) channels is essential for modulation of the cytosolic Ca(2+) ([Ca(2+)]C) signals, but such modulation may differ among cells. In chromaffin cells Ca(2+) entry through voltage-operated channels (VOCC) induces calcium release from the endoplasmic reticulum (ER) that amplifies the signal. [Ca(2+)]C microdomains as high as 20-50 µM are sensed by subplasmalemmal mitochondria, which accumulate large amounts of Ca(2+) through the mitochondrial Ca(2+) uniporter (MCU). Mitochondria confine the high-Ca(2+) microdomains (HCMD) to beneath the plasma membrane, where exocytosis of secretory vesicles happens. Cell core [Ca(2+)]C is much smaller (1-2 µM). By acting as a Ca(2+) sink, mitochondria stabilise the HCMD in space and time. In non-excitable HEK293 cells, activation of store-operated Ca(2+) entry (SOCE), triggered by ER Ca(2+) emptying, did also generate subplasmalemmal HCMD, but, in this case, most of the Ca(2+) was taken up by the ER rather than by mitochondria. The smaller size of the [Ca(2+)]C peak in this case (about 2 µM) may contribute to this outcome, as the SERCA has much higher Ca(2+) affinity than MCU. There is also possible that relative positioning of organelles, channels and effectors, as well as cytoskeleton and accessory proteins play an important role.The Journal of Physiology 06/2013; 592(2). DOI:10.1113/jphysiol.2013.255661 · 4.54 Impact Factor
Journal of the American College of Cardiology 02/1995; 25(2). DOI:10.1016/0735-1097(95)93150-B · 15.34 Impact Factor
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ABSTRACT: Eukaryotic cells are divided into distinct membrane-bound organelles with unique identities and specialized metabolic functions. Communication between organelles must take place to regulate the size, shape, and composition of individual organelles, as well as to coordinate transport between organelles. The endoplasmic reticulum (ER) forms an expansive membrane network that contacts and participates in crosstalk with several other organelles in the cell, most notably the plasma membrane (PM). ER-PM junctions have well-established functions in the movement of small molecules, such as lipids and ions, between the ER and PM. Recent discoveries have revealed additional exciting roles for ER-PM junctions in the regulation of cell signaling, ER shape and architecture, and PM domain organization.Current opinion in cell biology 03/2013; DOI:10.1016/j.ceb.2013.02.020 · 8.74 Impact Factor