Conditional knockout of Mn-SOD targeted to type IIB skeletal muscle fibers increases oxidative stress and is sufficient to alter aerobic exercise capacity

Lustgarten MS, Jang YC, Liu Y, Muller FL, Qi W, Steinhelper M, Brooks SV, Larkin L, Shimizu T, Shirasawa T, McManus LM, Bhattacharya A, Richardson A, Van Remmen H. Conditional knockout of Mn-SOD targeted to type IIB skeletal muscle fibers increases oxidative stress and is sufficient to alter aerobic exercise capacity. Am J Physiol Cell Physiol 297: C 1520-C1532, 2009. First published September 23, 2009; doi: 10.1152/ajpcell.00372.2009.--In vitro studies of isolated skeletal muscle have shown that oxidative stress is limiting with respect to contractile function. Mitochondria are a potential source of muscle function-limiting oxidants. To test the hypothesis that skeletal muscle-specific mitochondrial oxidative stress is sufficient to limit muscle function, we bred mice expressing Cre recombinase driven by the promoter for the inhibitory subunit of troponin (TnIFastiCre) with mice containing a floxed Sod2 (Sod[2.sup.fl/fl]) allele. Mn-SOD activity was reduced by 82% in glycolytic (mainly type II) muscle fiber homogenates from young TnIFastCreSod[2.sup.fl/fl] mice. Furthermore, Mn-SOD content was reduced by 70% only in type IIB muscle fibers. Aconitase activity was decreased by 56%, which suggests an increase in mitochondrial matrix superoxide. Mitochondrial superoxide release was elevated more than twofold by mitochondria isolated from glycolytic skeletal muscle in TnIFastCreSod[2.sup.fl/fl] mice. In contrast, the rate of mitochondrial [H.sub.2][O.sub.2] production was reduced by 33%, and only during respiration with complex II substrate. [F.sub.2]-isoprostanes were increased by 36% in tibialis anterior muscles isolated from TnIFastCreSod[2.sup.fl/fl] mice. Elevated glycolytic muscle-specific mitochondrial oxidative stress and damage in TnIFastCreSod[2.sup.fl/fl] mice were associated with a decreased ability of the extensor digitorum longus and gastrocnemius muscles to produce contractile force as a function of time, whereas force production by the soleus muscle was unaffected. TnIFastCreSod.[2.sup.fl/fl] mice ran 55% less distance on a treadmill than wild-type mice. Collectively, these data suggest that elevated mitochondrial oxidative stress and damage in glycolytic muscle fibers are sufficient to reduce contractile muscle function and aerobic exercise capacity. muscle function; contractile function; oxidative damage; free radical doi: 10.1152/ajpcell.00372.2009