Selected Publication:
Paltauf-Doburzynska, J; Frieden, M; Graier, WF. 
Mechanisms of Ca2+ store depletion in single endothelial cells in a Ca(2+)-free environment.
Cell Calcium. 1999; 25(5):345-353
 Doi: 10.1054/ceca.1999.0038
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PubMed
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- Leading authors Med Uni Graz
 
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Graier Wolfgang
 
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- Abstract:
 
- Depletion of agonist-sensitive Ca2+ stores results in activation of capacitative Ca2+ entry (CCE) in endothelial cells. The proportion of Ca2+ stores contributing to the regulation of CCE is unknown. In fura-2/am loaded single endothelial cells freshly isolated from bovine left circumflex coronary arteries, we investigated whether a resting period in a Ca(2+)-free environment results in emptying of bradykinin-sensitive Ca2+ stores (BsS) and activation of CCE. In a Ca(2+)-free environment, depletion of BsS occurred in a time-dependent manner (59% after 10 min in Ca(2+)-free solution). This effect was prevented by inhibition of the Na(+)-Ca2+ exchange but not by a blockade of ryanodine-sensitive Ca2+ release (RsCR). In contrast to BsS, mitochondrial Ca2+ content remained unchanged in the Ca(2+)-free environment. Remarkably, activity of CCE (monitored as Mn2+ influx) did not increase after depletion of BsS in the Ca(2+)-free environment. In contrast to Mn2+ influx, the effect of re-addition of Ca2+ to elevate bulk Ca2+ concentration ([Ca2+]b) decreased with the time the cells rested in Ca(2+)-free buffer. This decrease was prevented by an inhibition of RsCR. In low Na+ conditions the effect of Ca2+ on [Ca2+]b was reduced while it did not change the time the cells rested in Ca(2+)-free solution. After a 2 min period in low Na+ conditions, ryanodine-induced Ca2+ extrusion was markedly diminished. Inhibition of RsCR re-established the effect of Ca2+ on [Ca2+]b in low Na+ conditions. Collapsing subplasmalemmal Ca2+ stores with nocodazole, increased the effect of Ca2+ on [Ca2+]b. In nocodazole-treated cells, the effect of Ca2+ on [Ca2+]b was not reduced in Ca(2+)-free environment. These data indicate that activation of CCE is not associated with the agonist-sensitive Ca2+ pools that deplete rapidly in a Ca(2+)-free environment. Subplasmalemmal ryanodine-sensitive Ca2+ stores (RsS) are emptied in Ca(2+)-free/low Na+ solution and re-sequester Ca2+ which enters the cells prior an increase in [Ca2+]b occurs. Thus, in endothelial cells there are differences in the functions of various subplasmalemmal Ca2+ stores (i.e. BsS and RsS), which include either activation of CCE or regulation of subplasmalemmal Ca2+.
 
- Find related publications in this database (using NLM MeSH Indexing)
 
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Animals -  
 
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Bradykinin - pharmacology 
 
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Calcium - metabolism 
 
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Calcium Channels - drug effects 
 
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Cattle - drug effects 
 
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Cells, Cultured - drug effects 
 
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Coronary Vessels - metabolism 
 
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Endoplasmic Reticulum - metabolism 
 
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Endothelium, Vascular - metabolism 
 
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Kinetics - metabolism 
 
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Magnesium - metabolism 
 
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Models, Biological - metabolism 
 
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Nocodazole - pharmacology 
 
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Ryanodine - pharmacology 
 
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Sodium-Calcium Exchanger - physiology 
 
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Spectrometry, Fluorescence - physiology 
 
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Time Factors - physiology