Document Detail


Regulation of calcineurin activity in insulin-secreting cells: stimulation by Hsp90 during glucocorticoid-induced apoptosis.
MedLine Citation:
PMID:  18611438     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previously, we described that apoptotic cell death induced by the synthetic glucocorticoid dexamethasone (dex) is inhibited by calcineurin inhibitors, FK506 and deltamethrin, in insulin-secreting cells. The aim of the present study was to examine the mechanism of dex-dependent activation of calcineurin. In INS-1 cells cultured up to 4d with dex (100 nmol/l), the percentage of apoptosis, quantified by condensed nuclei and TUNEL positive cells, increased from 1% to 10.9%. FK506 inhibited dex-mediated cell death. Apoptosis was significantly higher at glucose concentrations that induce [Ca(2+)](i) oscillations than at low, non-stimulatory glucose. Dex had no acute effect on [Ca(2+)](i). Calcineurin activity, measured in control and dex-treated cell homogenates, revealed that maximal activity and the sensitivity to the substrate RII peptide was unaltered. However, dex treatment significantly increased enzyme activity at submaximal, physiological Ca(2+) concentrations. Dex did not stimulate the Ca(2+)-dependent protease calpain, known to activate calcineurin by cleavage, as no cleaved calcineurin was detectable. Furthermore, the calpain inhibitor ALLN did not counteract dex-dependent cell death. Western blotting revealed that in dex-treated cells heat shock protein 90 (Hsp90), a component of the glucocorticoid receptor (GR) known to stimulate calcineurin, was increased while calcineurin protein levels were unchanged. In immunoprecipitates with calcineurin antibodies, Hsp90 was only detected in dex-treated cell homogenates. These data suggest that dex-induced apoptosis involves release of Hsp90 from the stimulated GR complex, subsequent binding to and activation of calcineurin, that may contribute to dex-mediated cell death in the presence of high glucose.
Authors:
Felicia Ranta; Martina Düfer; Björn Stork; Sebastian Wesselborg; Gisela Drews; Hans-Ulrich Häring; Florian Lang; Susanne Ullrich
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-06-18
Journal Detail:
Title:  Cellular signalling     Volume:  20     ISSN:  0898-6568     ISO Abbreviation:  Cell. Signal.     Publication Date:  2008 Oct 
Date Detail:
Created Date:  2008-08-25     Completed Date:  2008-11-14     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8904683     Medline TA:  Cell Signal     Country:  England    
Other Details:
Languages:  eng     Pagination:  1780-6     Citation Subset:  IM    
Affiliation:
Institute of Physiology, University of Tübingen, Gmelinstrasse 5, D-72076 Tübingen, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / drug effects*
Calcineurin / antagonists & inhibitors,  metabolism*
Calcium / pharmacology
Calpain / antagonists & inhibitors
Cytoplasm / drug effects,  metabolism
Dexamethasone / pharmacology*
Drug Synergism
Enzyme Activation / drug effects
Fura-2
Glucose / pharmacology
HSP90 Heat-Shock Proteins / metabolism*
Insulin-Secreting Cells / cytology*,  drug effects,  enzymology*
Protein Binding / drug effects
Rats
Substrate Specificity / drug effects
Tacrolimus / pharmacology
Chemical
Reg. No./Substance:
0/HSP90 Heat-Shock Proteins; 109581-93-3/Tacrolimus; 50-02-2/Dexamethasone; 50-99-7/Glucose; 7440-70-2/Calcium; 96314-98-6/Fura-2; EC 3.1.3.16/Calcineurin; EC 3.4.22.-/Calpain

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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