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The effects of various concentrations of thapsigargin, a specific inhibitor of Ca2+ -ATPase in the endoplasmic reticulum (ER) membrane, on calcium homeostasis in lymphoidal T cells (Jurkat) were investigated. Preincubation of these cells suspended in nominally calcium-free medium with 0.1 UM thapsigargin resulted in a complete release of Ca2+ from intracellular calcium stores. When the medium was supplemented with 3 mM CaCl2 the cells maintained constantly elevated level of cytosolic Ca2+ . However, thapsigargin applied at lower concentration produced only a partial depletion of the stores. For example, in the cells pretreated with 1 nM thapsigargin and suspended in calcium-free medium approximately 75% of the calcium content was released from the intracellular stores. The addition of 3 mM CaCl2 to such cell suspension led to a transient increase in cytosolic calcium concentration, followed by a return to a lower steady-state. This phenomenon, related to the refilling of the ER by Ca2+ , allowed to estimate the half-time for the process of cell recovery after activation of store-operated calcium channels. By this approach we have found that carbonyl cyanide m-chlorophenylhydrazone, which has been documented to inhibit calcium entry into Jurkat cells, does not influence the stability of the intracellular signal involved in the activation of store-operated calcium channels.
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Opis fizyczny
p.157-161,fig.
Twórcy
autor
- Nencki Institute of Experimental Biology, L.Pasteura 3, 02-093 Warsaw, Poland
autor
autor
Bibliografia
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- 15.Wasniewska, M. & Duszynski, J. (2000) The role of mitochondrial dysfunction in regulation of store-operated calcium channels in glioma C6 and human fibroblast cells. FEBS Lett. 478, 237-240.
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Bibliografia
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