Microtubule alteration is an early cellular reaction to the metabolic challenge in ischemic cardiomyocytes

Cytoskeleton damage, particularly microtubule (MT) alterations, may play an important role in the pathogenesis of ischemia-induced myocardial injury. However, this disorganization has been scarcely confirmed in the cellular context. We evaluated MT network disassembly in myoblast cell line H9c2 and...

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Bibliographic Details
Published in:Molecular and cellular biochemistry 2004-03, Vol.258 (1-2), p.99-108
Main Authors: Vandroux, David, Schaeffer, Céline, Tissier, Cindy, Lalande, Alain, Bès, Sandrine, Rochette, Luc, Athias, Pierre
Format: Article
Language:English
Subjects:
Animals
Animals, Newborn
Cardiomyocytes
Cell Survival
Cells
Cells, Cultured
Computer Science
Hypoxia
Ischemia
Medical Imaging
Microtubules - metabolism
Myocardial Infarction - metabolism
Myocardial Infarction - pathology
Myocytes, Cardiac - metabolism
Myocytes, Cardiac - pathology
Rats
Tubulin - metabolism
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Summary:Cytoskeleton damage, particularly microtubule (MT) alterations, may play an important role in the pathogenesis of ischemia-induced myocardial injury. However, this disorganization has been scarcely confirmed in the cellular context. We evaluated MT network disassembly in myoblast cell line H9c2 and in neonatal rat cardiomyocytes in an in vitro substrate-free hypoxia model of simulated ischemia (SI). After different duration of SI from 30 up to 180 min, the cells were fixed and the microtubule network was revealed by immunocytochemistry. The microtubule alterations were quantified using a house-developed image analysis program. Additionally, the tubulin fraction were extracted and quantified by Western blotting. The cell respiration, the release of cellular LDH and the cell viability were evaluated at the same periods. An early MT disassembly was observed after 60 min of SI. The decrease in MT fluorescence intensity at 60 and 90 min was correlated with a microtubule disassembly. Conversely, SI-induced significant LDH release (35%) and decrease in cell viability (34%) occurred after 120 min only. These results suggest that the simulated ischemia-induced changes in MT network should not be considered as an ultrastructural hallmark of the cell injury and could rather be an early ultrastructural correlate of the cellular reaction to the metabolic challenge.
ISSN:0300-8177
1573-4919
DOI:10.1023/B:MCBI.0000012840.67616.cc