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Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl−
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Zeitschriftentitel: | American Journal of Physiology-Renal Physiology |
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Personen und Körperschaften: | , , , , , , , , , |
In: | American Journal of Physiology-Renal Physiology, 286, 2004, 6, S. F1046-F1053 |
Format: | E-Article |
Sprache: | Englisch |
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American Physiological Society
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author_facet |
Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe |
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author |
Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe |
spellingShingle |
Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe American Journal of Physiology-Renal Physiology Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− Physiology |
author_sort |
lang, karl s. |
spelling |
Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe 1931-857X 1522-1466 American Physiological Society Physiology http://dx.doi.org/10.1152/ajprenal.00263.2003 <jats:p>Osmotic shock by addition of sucrose to the medium stimulates erythrocyte sphingomyelinase with subsequent ceramide formation and triggers Ca<jats:sup>2+</jats:sup>entry through stimulation of cation channels. Both ceramide and Ca<jats:sup>2+</jats:sup>activate an erythrocyte scramblase, leading to breakdown of phosphatidylserine asymmetry, a typical feature of apoptosis. Because erythrocytes are regularly exposed to osmotic shock during passage of kidney medulla, the present study explored the influence of NaCl and urea on erythrocyte phosphatidylserine exposure as determined by annexin binding. The percentage of annexin-binding erythrocytes increased from <5 to 80 ± 4% ( n = 4) upon addition of 650 mM sucrose, an effect paralleled by activation of the cation channel and stimulation of ceramide formation. The number of annexin-binding erythrocytes increased only to 18% after addition of 325 mM NaCl and was not increased by addition of 650 mM urea. According to whole cell patch-clamp experiments, the cation conductance was activated by replacement of extracellular Cl<jats:sup>−</jats:sup>with gluconate at isotonic conditions or by addition of hypertonic sucrose or urea. Although stimulating the cation conductance, urea abrogated the annexin binding and concomitant increase of ceramide levels induced by osmotic cell shrinkage. In vitro sphingomyelinase assays demonstrated a direct inhibitory effect of urea on sphingomyelinase activity. Urea did not significantly interfere with annexin binding after addition of ceramide. In conclusion, both Cl<jats:sup>−</jats:sup>and urea blunt erythrocyte phosphatidylserine exposure after osmotic shock. Whereas Cl<jats:sup>−</jats:sup>is effective through inhibition of the cation conductance, urea exerts its effect through inhibition of sphingomyelinase, thus blunting formation of ceramide.</jats:p> Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl<sup>−</sup> American Journal of Physiology-Renal Physiology |
doi_str_mv |
10.1152/ajprenal.00263.2003 |
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American Physiological Society |
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American Journal of Physiology-Renal Physiology |
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title |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_unstemmed |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_full |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_fullStr |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_full_unstemmed |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_short |
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_sort |
inhibition of erythrocyte phosphatidylserine exposure by urea and cl<sup>−</sup> |
topic |
Physiology |
url |
http://dx.doi.org/10.1152/ajprenal.00263.2003 |
publishDate |
2004 |
physical |
F1046-F1053 |
description |
<jats:p>Osmotic shock by addition of sucrose to the medium stimulates erythrocyte sphingomyelinase with subsequent ceramide formation and triggers Ca<jats:sup>2+</jats:sup>entry through stimulation of cation channels. Both ceramide and Ca<jats:sup>2+</jats:sup>activate an erythrocyte scramblase, leading to breakdown of phosphatidylserine asymmetry, a typical feature of apoptosis. Because erythrocytes are regularly exposed to osmotic shock during passage of kidney medulla, the present study explored the influence of NaCl and urea on erythrocyte phosphatidylserine exposure as determined by annexin binding. The percentage of annexin-binding erythrocytes increased from <5 to 80 ± 4% ( n = 4) upon addition of 650 mM sucrose, an effect paralleled by activation of the cation channel and stimulation of ceramide formation. The number of annexin-binding erythrocytes increased only to 18% after addition of 325 mM NaCl and was not increased by addition of 650 mM urea. According to whole cell patch-clamp experiments, the cation conductance was activated by replacement of extracellular Cl<jats:sup>−</jats:sup>with gluconate at isotonic conditions or by addition of hypertonic sucrose or urea. Although stimulating the cation conductance, urea abrogated the annexin binding and concomitant increase of ceramide levels induced by osmotic cell shrinkage. In vitro sphingomyelinase assays demonstrated a direct inhibitory effect of urea on sphingomyelinase activity. Urea did not significantly interfere with annexin binding after addition of ceramide. In conclusion, both Cl<jats:sup>−</jats:sup>and urea blunt erythrocyte phosphatidylserine exposure after osmotic shock. Whereas Cl<jats:sup>−</jats:sup>is effective through inhibition of the cation conductance, urea exerts its effect through inhibition of sphingomyelinase, thus blunting formation of ceramide.</jats:p> |
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author | Lang, Karl S., Myssina, Swetlana, Lang, Philipp A., Tanneur, Valerie, Kempe, Daniela S., Mack, Andreas F., Huber, Stephan M., Wieder, Thomas, Lang, Florian, Duranton, Christophe |
author_facet | Lang, Karl S., Myssina, Swetlana, Lang, Philipp A., Tanneur, Valerie, Kempe, Daniela S., Mack, Andreas F., Huber, Stephan M., Wieder, Thomas, Lang, Florian, Duranton, Christophe, Lang, Karl S., Myssina, Swetlana, Lang, Philipp A., Tanneur, Valerie, Kempe, Daniela S., Mack, Andreas F., Huber, Stephan M., Wieder, Thomas, Lang, Florian, Duranton, Christophe |
author_sort | lang, karl s. |
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container_title | American Journal of Physiology-Renal Physiology |
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description | <jats:p>Osmotic shock by addition of sucrose to the medium stimulates erythrocyte sphingomyelinase with subsequent ceramide formation and triggers Ca<jats:sup>2+</jats:sup>entry through stimulation of cation channels. Both ceramide and Ca<jats:sup>2+</jats:sup>activate an erythrocyte scramblase, leading to breakdown of phosphatidylserine asymmetry, a typical feature of apoptosis. Because erythrocytes are regularly exposed to osmotic shock during passage of kidney medulla, the present study explored the influence of NaCl and urea on erythrocyte phosphatidylserine exposure as determined by annexin binding. The percentage of annexin-binding erythrocytes increased from <5 to 80 ± 4% ( n = 4) upon addition of 650 mM sucrose, an effect paralleled by activation of the cation channel and stimulation of ceramide formation. The number of annexin-binding erythrocytes increased only to 18% after addition of 325 mM NaCl and was not increased by addition of 650 mM urea. According to whole cell patch-clamp experiments, the cation conductance was activated by replacement of extracellular Cl<jats:sup>−</jats:sup>with gluconate at isotonic conditions or by addition of hypertonic sucrose or urea. Although stimulating the cation conductance, urea abrogated the annexin binding and concomitant increase of ceramide levels induced by osmotic cell shrinkage. In vitro sphingomyelinase assays demonstrated a direct inhibitory effect of urea on sphingomyelinase activity. Urea did not significantly interfere with annexin binding after addition of ceramide. In conclusion, both Cl<jats:sup>−</jats:sup>and urea blunt erythrocyte phosphatidylserine exposure after osmotic shock. Whereas Cl<jats:sup>−</jats:sup>is effective through inhibition of the cation conductance, urea exerts its effect through inhibition of sphingomyelinase, thus blunting formation of ceramide.</jats:p> |
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institution | DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
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spelling | Lang, Karl S. Myssina, Swetlana Lang, Philipp A. Tanneur, Valerie Kempe, Daniela S. Mack, Andreas F. Huber, Stephan M. Wieder, Thomas Lang, Florian Duranton, Christophe 1931-857X 1522-1466 American Physiological Society Physiology http://dx.doi.org/10.1152/ajprenal.00263.2003 <jats:p>Osmotic shock by addition of sucrose to the medium stimulates erythrocyte sphingomyelinase with subsequent ceramide formation and triggers Ca<jats:sup>2+</jats:sup>entry through stimulation of cation channels. Both ceramide and Ca<jats:sup>2+</jats:sup>activate an erythrocyte scramblase, leading to breakdown of phosphatidylserine asymmetry, a typical feature of apoptosis. Because erythrocytes are regularly exposed to osmotic shock during passage of kidney medulla, the present study explored the influence of NaCl and urea on erythrocyte phosphatidylserine exposure as determined by annexin binding. The percentage of annexin-binding erythrocytes increased from <5 to 80 ± 4% ( n = 4) upon addition of 650 mM sucrose, an effect paralleled by activation of the cation channel and stimulation of ceramide formation. The number of annexin-binding erythrocytes increased only to 18% after addition of 325 mM NaCl and was not increased by addition of 650 mM urea. According to whole cell patch-clamp experiments, the cation conductance was activated by replacement of extracellular Cl<jats:sup>−</jats:sup>with gluconate at isotonic conditions or by addition of hypertonic sucrose or urea. Although stimulating the cation conductance, urea abrogated the annexin binding and concomitant increase of ceramide levels induced by osmotic cell shrinkage. In vitro sphingomyelinase assays demonstrated a direct inhibitory effect of urea on sphingomyelinase activity. Urea did not significantly interfere with annexin binding after addition of ceramide. In conclusion, both Cl<jats:sup>−</jats:sup>and urea blunt erythrocyte phosphatidylserine exposure after osmotic shock. Whereas Cl<jats:sup>−</jats:sup>is effective through inhibition of the cation conductance, urea exerts its effect through inhibition of sphingomyelinase, thus blunting formation of ceramide.</jats:p> Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl<sup>−</sup> American Journal of Physiology-Renal Physiology |
spellingShingle | Lang, Karl S., Myssina, Swetlana, Lang, Philipp A., Tanneur, Valerie, Kempe, Daniela S., Mack, Andreas F., Huber, Stephan M., Wieder, Thomas, Lang, Florian, Duranton, Christophe, American Journal of Physiology-Renal Physiology, Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl−, Physiology |
title | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_full | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_fullStr | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_full_unstemmed | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_short | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
title_sort | inhibition of erythrocyte phosphatidylserine exposure by urea and cl<sup>−</sup> |
title_unstemmed | Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl− |
topic | Physiology |
url | http://dx.doi.org/10.1152/ajprenal.00263.2003 |