Partial hexokinase II knockout results in acute ischemia-reperfusion damage in skeletal muscle of male, but not female, mice

Cellular studies have demonstrated a protective role of mitochondrial hexokinase against oxidative insults. It is unknown whether HK protective effects translate to the in vivo condition. In the present study, we hypothesize that HK affects acute ischemia–reperfusion injury in skeletal muscle of the intact animal. Male and female heterozygote knockout HKII (HK(+/-)), heterozygote overexpressed HKII (HK(tg)), and their wild-type (WT) C57Bl/6 littermates mice were examined. In anesthetized animals, the left gastrocnemius medialis (GM) muscle was connected to a force transducer and continuously stimulated (1-Hz twitches) during 60 min ischemia and 90 min reperfusion. Cell survival (%LDH) was defined by the amount of cytosolic lactate dehydrogenase (LDH) activity still present in the reperfused GM relative to the contralateral (non-ischemic) GM. Mitochondrial HK activity was 72.6 ± 7.5, 15.7 ± 1.7, and 8.8 ± 0.9 mU/mg protein in male mice, and 72.7 ± 3.7, 11.2 ± 1.4, and 5.9 ± 1.1 mU/mg in female mice for HK(tg), WT, and HK(+/-), respectively. Tetanic force recovery amounted to 33 ± 7% for male and 17 ± 4% for female mice and was similar for HK(tg), WT, and HK(+/-). However, cell survival was decreased (p = 0.014) in male HK(+/-) (82 ± 4%LDH) as compared with WT (98 ± 5%LDH) and HK(tg) (97 ± 4%LDH). No effects of HKII on cell survival was observed in female mice (92 ± 2% LDH). In conclusion, in this mild model of acute in vivo ischemia–reperfusion injury, a partial knockout of HKII was associated with increased cell death in male mice. The data suggest for the first time that HKII mediates skeletal muscle ischemia–reperfusion injury in the intact male animal

Effects of physiological amounts of high- and low-rate chronic stimulation on fast-twitch muscle of the cat hindlimb. II. Endurance-related properties

1. Long-term electrical stimulation was given to the peroneal nerve of deafferented hindlimbs in hemispinalized adult cats. The amount of stimulation covered 0.5-5.5% of total time per day, different in different animals. For some aspects of the present study, use was also made of cats subjected to "tonic" patterns of chronic stimulation (typically covering 50% of total time; 10, 16). 2. In a terminal acute experiment under general anesthesia, performed after 4 or 8 wk of long-term stimulation, one of the treated peroneal muscles (m. peroneus longus, PerL) was used for measurements of the resistance to contractile fatigue. The fatigue test consisted of 0.33-s bursts of motor-nerve stimulation at 40 Hz, repeated once a second for 4 min (6, 7). During this fatigue test, the evoked compound spikes of the muscle were recorded by electromyographic (EMG) techniques. Following the physiological procedures, PerL was removed for further histochemical analysis. In transverse sections, measurements of optical density were made in central regions of single fibers after staining for the activity of an oxidative enzyme, succinate dehydrogenase (core SDH). 3. Findings from chronically stimulated PerL muscles were compared with three kinds of control PerL muscles: 1) those contralateral to the stimulated ones, 2) those from the operated side of animals that had been deafferented and hemispinalized but not subjected to chronic stimulation, and 3) those from untreated normal animals. 4. Stimulation patterns covering both greater than or equal to 50% and 5-5.5% of daily time gave a marked improvement of fatigue resistance. Pulse rate seemed of little importance for these effects. The pattern covering only 0.5% of total daily time caused no increase of contractile endurance beyond that of normal muscles. 5. During the fatigue test of a control muscle (see above), the amplitude of the compound EMG spikes typically showed a marked decline. This "EMG depression" was effectively counteracted by all the present patterns of chronic stimulation, including the 0.5% pattern. 6. Fibers of chronically stimulated muscles became more similar to each other with respect to their density of core SDH staining. However, among muscles treated during 0.5-5.5% of total daily time, the degree and pattern of change in core SDH staining was not related to the amount and pattern of chronic stimulation or to the resulting degree of contractile enduranc

Mitochondrial hexokinase and cardioprotection of the intact heart

The interaction of hexokinase with mitochondria has emerged as a powerful mechanism in protecting many cell types against cell death. However, the role of mitochondrial hexokinase (mitoHK) in cardiac ischemia-reperfusion injury has as of yet received little attention. In this review we examine whether increased binding of hexokinase to the mitochondrion is also an integral component of cardioprotective signalling. We discuss observations in cardiac mitochondrial activation that directed us to the hypothesis of hexokinase cellular redistribution with reversible, cardioprotective ischemia, summarize the data showing that many cardioprotective interventions, such as ischemic preconditioning, insulin, morphine and volatile anesthetics, increase mitochondrial hexokinase binding within the intact heart, and discuss similarities between mitochondrial hexokinase association and ischemic preconditioning. Although most data indicate that mitochondrial hexokinase may indeed be an integral part of cardioprotection, a definitive proof for a causal relation between the amount of mitoHK and cardiac ischemia-reperfusion injury in the intact heart is eagerly awaited. When such relationship is indeed observed, the association of hexokinase with mitochondria will offer an opportunity to develop new therapies to combat ischemic cardiac disease