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Received June 11, 2004
Revised July 1, 2004
Accepted after revision August 25, 2004
1 University of Liverpool
2 Department of Medicine
* To whom correspondence should be addressed. E-mail: m.j.jackson{at}liverpool.ac.uk.
Adaptations of skeletal muscle following exercise are accompanied by changes in gene expression, which can result in protection against subsequent potentially damaging exercise. One cellular signal activating these adaptations may be an increased production of reactive oxygen and nitrogen species (ROS). The aim of this study was to examine the effect of a short period of non-damaging contractions on the subsequent susceptibility of muscle to contraction-induced damage and to examine the changes in gene expression that occur following the initial contraction protocol. Comparisons with changes in gene expression in cultured myotubes following treatment with a non-damaging concentration of hydrogen peroxide (H2O2) were used to identify redox-sensitive genes whose expression may be modified by the increased ROS production during contractions. Hindlimb muscles of mice were subjected to a pre-conditioning, non-damaging isometric contraction protocol in vivo. Four or 12 hours later, extensor digitorum longus (EDL) and soleus muscles were removed and subjected to a (normally) damaging contraction protocol in vitro. Muscles were also analysed for changes in gene expression induced by the preconditioning protocol using cDNA expression techniques. In a parallel study, C2C12 myotubes were treated with a non-damaging concentration (100mM) of H2O2 and, at 4 and 12 hours following treatment, myotubes were treated with a damaging concentration of H2O2 (2mM). Myotubes were analysed for changes in gene expression at 4 hours following treatment with 100mM H2O2 alone. Data demonstrate that a prior period of non-damaging contractile activity resulted in significant protection of EDL and soleus muscles against a normally damaging contraction protocol 4 hours later. This protection was associated with significant changes in gene expression. Prior treatment of myotubes with a non-damaging concentration of H2O2 also resulted in significant protection against a damaging treatment, 4 and 12 hours later. Comparison of changes in gene expression in both studies identified heme oxygenase-1 as the sole gene showing increased expression during adaptation in both instances suggesting that activation of this gene results from the increased ROS production during contractile activity and that it may play a role in protection of muscle cells against subsequent exposure to damaging activity.
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