Preferences help
enabled [disable] Abstract
Number of results
2001 | 48 | 1 | 157-161
Article title

Mitochondrial uncoupling does not influence the stability of the intracellular signal activating plasma membrane calcium channels.

Title variants
Languages of publication
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 μM 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.

Physical description
  • Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warszawa, Poland
  • Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warszawa, Poland
  • Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warszawa, Poland
  • Barrit, G.J. (1999) Receptor-activated Ca2+ inflow in animal cells: A variety of pathways tailored to meet different intracellular Ca2+ signalling requirements. Biochem. J. 337, 153-169.
  • Berridge, M.J. (1995) Capacitative calcium entry. Biochem. J. 312, 1-11.
  • Fernando, C.K., Gregory, R.B. & Barritt, G.J. (1998) Protein kinase A regulates the disposition of Ca2+ which enters the cytoplasmic space through store- activated Ca2+ channels in rat hepatocytes by diverting inflowing Ca2+ to mitochondria. Biochem. J. 267, 877-884.
  • Gamberucci, A., Innocenti, B., Fulceri, R., Banhegy, G., Giunti, R. & Benedetti, A. (1994) Modulation of Ca2+ influx dependent on store depletion by intracellular adenine-guanine nucleotide levels. J. Biol. Chem. 269, 23597-23602.
  • Grynkiewicz, G., Poenie, M. & Tsien, R.Y. (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440-3450.
  • Hofer, A.M., Fasolato, C. & Pozzan, T. (1998) Capacitative Ca2+ entry is closely linked to the filling state of internal Ca2+ stores: A study using simultaneous measurements of ICRAC and intraluminal [Ca2+]. J. Cell Biol. 140, 325-334.
  • Hoth, M., Fanger, C.M. & Lewis, R.S. (1997) Mitochondrial regulation of store-operated calcium signaling in T lymphocytes. J. Cell Biol. 137, 633-648.
  • Kiselyov, K., Xu, X., Mozhayeva, G., Kuo, T., Pessah, I., Mignery, G., Zhu, X., Birnbaumer, L. & Muallem, S. (1998) Functional interaction between InsP3 receptors and store-operated Htrp3 channels. Nature 396, 478-482.
  • Makowska, A., Zabłocki, K. & Duszyński, J. (2000) the role of mitochondria in the regulation of calcium influx into Jurkat cells. Eur. J. Biochem. 267, 877-884.
  • Parekh, A.B. (1998) Slow feedback inhibition of calcium release-activated calcium current by calcium entry. J. Biol. Chem. 273, 14925-14932.
  • Parekh, A.B. & Penner, R. (1997) Store depletion and calcium influx. Physiol. Rev. 77, 901-929.
  • Putney, J.W. (1986) A model for receptor-regulated calcium entry. Cell Calcium 7, 1-12.
  • Putney, J.W. (1990) Capacitative calcium entry revisited. Cell Calcium 11, 611-624.
  • Thastrup, O., Dawson, A.P., Scharff, O., Foder, B., Cullen, P.J. Drobak, B.K., Bjerrum, P.J., Christensen, S.B. & Hanley, M.R. (1994) Thapsigargin, a novel molecular probe for studying intracellular calcium release and storage. Agents Actions 43, 187-193.
  • Waśniewska, M. & Duszyński, 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.
  • Zweifach, A. & Lewis, R.S. (1995a) Slow calcium-dependent inactivation of depletion-activated calcium current. Store-dependent and -independent mechanisms. J. Biol. Chem. 270, 14445- 14451.
  • Zweifach, A. & Lewis, R.S. (1995b) Rapid inactivation of depletion-activated calcium current (ICRAC) due to local calcium feedback. J. Gen. Physiol. 105, 209-226.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.