Do changes in mitochondrial quality contribute to increases in skeletal muscle oxidative capacity following endurance training?

Abstract

Endurance training improves skeletal muscle oxidative capacity, but the molecular adaptations that drive this process are not fully understood. Specifically, whether oxidative capacity is improved solely by augmentations in mitochondrial quantity, or by enhanced mitochondrial quality as well, is unclear. For example, compared to active individuals, elite endurance athletes exhibit superior in vitro oxidative capacity even when normalised to citrate synthase (CS) activity, a common marker of mitochondrial quantity (Jacobs & Lundby, 2013). Two important questions emerge from these results: (1) what molecular or enzymatic aspects of mitochondrial composition might allow rates of in vitro oxidative capacity to increase for a given mitochondrial volume, and (2) what endurance training methods are best suited to stimulate these positive adaptations? Adaptations to endurance training have traditionally been studied following a period of moderate intensity continuous training (MICT), which is characterised by prolonged periods of aerobic activity at submaximal workloads (i.e. high volume training). More recently, interest has shifted towards high intensity interval training (HIIT), which involves repeated bouts of vigorous-intensity exercise interspersed with periods of recovery. Daussin et al. (2008) reported that in vitro oxidative capacity is significantly improved by HIIT, but not by MICT. Larsen et al. (2013) demonstrated that as few as six sessions of HIIT are sufficient to improve in vivo markers of skeletal muscle oxidative capacity, measured as the maximal rate of phosphocreatine resynthesis. However, these studies did not “normalise” oxidative capacity measurements to mitochondrial quantity, nor did they measure changes in mitochondrial enzymatic composition, making it difficult to infer the molecular mechanisms that control measures of oxidative capacity and how training intensity may influence these changes. In an article in The Journal of Physiology, MacInnis et al. (2016) attempted to address this gap in the literature by comparing changes in whole muscle and mitochondria-specific in vitro oxidative capacity after 2 weeks of MICT and HIIT. To compare the different training modalities, they used single-leg cycle ergometry, which allowed all participants (n=10 young males) to perform both MICT and HIIT over the same training period and serve as their own controls. Peak aerobic capacity (Wpeak) was measured on each leg using a ramp protocol before and after 2 weeks of endurance training. Participants completed six sessions of single-leg MICT (30 min at 50% Wpeak) and HIIT (4 bouts of 5 min at 65% Wpeak and 2.5 min at 20% Wpeak). Muscle biopsies were taken from the vastus lateralis of each leg preand post-training to measure markers of mitochondrial composition and mitochondrial oxidative capacity. Markers of mitochondrial composition included CS (used as a marker for mitochondrial quantity), cytochrome c oxidase subunit 4 (COXIV), NADH:ubiquinone oxidoreductase subunit A9 (NDUFA9), and mitofusin 2 (MFN2); the protein content of the latter three was measured in myosin heavy chain (MHC) I and IIA fibres. Oxidative capacity of permeabilised muscle fibres was measured in vitro using a substrate uncoupler inhibitor titration protocol that allowed maximal O2-respiratory rates (J O2 ) through complexes I and II of the electron transport chain to be determined separately. Mass-specific J O2 was calculated as J O2 /muscle biopsy mass, whereas mitochondria-specific J O2 was calculated as the mass-specific J O2 normalised to CS content. The increase in whole muscle CS activity was significantly greater following HIIT compared to MICT (+39% vs. +11%, respectively). HIIT also produced significantly greater improvements in mass-specific J O2 through complex I (HIIT +22% vs. MICT –7%) and complex I+II (HIIT +22% vs. MICT –9%). In contrast, neither training method stimulated improvements in mitochondria-specific J O2 . And although MICT and HIIT both stimulated increases in COXIV, NDUFA9 and MFN2 protein content, none of these increases appeared to be fibre-type specific. Consistent with the authors’ hypotheses, HIIT was more effective than MICT for improving skeletal muscle mitochondrial quantity (i.e. CS activity), which coincided with greater increases in mass-specific J O2 . However, neither training method stimulated significant improvements in mitochondria-specific J O2 (i.e. mitochondrial quality). Based on these observations, it is possible that endurance training stimulates changes in mitochondrial abundance prior to mitochondrial quality. Alternatively stated, perhaps improvements in mitochondrial quality only occur once mitochondrial abundance has increased to the limit allowed by the spatial constraints of the muscle fibre. It would be interesting to track these changes incrementally over the course of several weeks to see if changes in massand mitochondria-specific J O2 are time-dependent. We were also intrigued that training method did not differentially impact changes in COXIV, NDUFA9 and MFA2 protein content within MHC I and IIA muscles fibres. Although previous studies have shown that six sessions of HIIT are sufficient to produce improvements in skeletal muscle oxidative capacity (Daussin et al. 2008; Larsen et al. 2013), the on-interval exercise intensity in these studies has typically been much greater than the 65% of Wpeak employed by MacInnis et al. (2016). Thus, the relatively modest difference in training intensity prescribed for HIIT and MICT (65 vs. 50% Wpeak, respectively) may have been insufficient to elicit different energetic overloads between the two training methods. Perhaps increasing the intensity of the HIIT protocol (e.g. 80% Wpeak) and shortening the on-interval duration would have elicited different responses between the two training protocols. It is also possible that enzymes other than those of the electron transport chain are limiting to J O2 and oxidative capacity. For example, during in vitro respirometry, mitochondrial J O2 is often stimulated with large, saturating boluses of ADP. Under these conditions, maximal J O2 is essentially limited by the rate of ADP transport