William M. Southern

Activity-induced changes in skeletal muscle metabolism measured with optical spectroscopy.

PURPOSE Previous studies have used near-infrared spectroscopy (NIRS) to measure skeletal muscle mitochondrial capacity. This study tested the hypothesis that NIRS-measured mitochondrial capacity would improve with endurance exercise training and decline with detraining. METHODS Nine young participants performed 4 wk of progressively increasing endurance exercise training of the wrist flexor muscles followed by approximately 5 wk of inactivity. The rate of recovery of muscle oxygen consumption (mV(˙)O₂) was measured with NIRS every 3-7 d, indicating mitochondrial oxidative capacity. RESULTS A linear increase in mitochondrial capacity (NIRS rate constant) was found with a group average of 64% ± 37% improvement after 4 wk of exercise training (P < 0.05). Mitochondrial capacity declined exponentially upon cessation of exercise training, with a mean half-time of approximately 7.7 d. CONCLUSIONS Both the magnitude and the time course of mitochondrial adaptations to exercise training and detraining measured with NIRS was consistent with previous studies using both in vitro and in vivo techniques. These findings show that NIRS-based measurements can detect meaningful changes in mitochondrial capacity.

Reproducibility of near-infrared spectroscopy measurements of oxidative function and postexercise recovery kinetics in the medial gastrocnemius muscle.

The purpose of this study was to assess the reproducibility of resting blood flow, resting oxygen consumption, and mitochondrial capacity in skeletal muscle using near-infrared spectroscopy (NIRS). We also determined the influence of 2 exercise modalities (ergometer and rubber exercise bands) on the NIRS measurements. Fifteen young, healthy participants (5 female, 10 male) were tested on 2 nonconsecutive occasions within an 8-day period. The NIRS device was placed on the medial gastrocnemius. Venous and arterial occlusions were performed to obtain blood flow and oxygen consumption. A series of repeated arterial occlusions was used to measure the recovery kinetics of muscle oxygen consumption after ∼7-10 s of voluntary plantar flexion exercise. Resting blood flow had mean coefficients of variation (CV) of 42% and 38% for bands and ergometer, respectively, and resting metabolism had mean CVs of 17% and 12% for bands and ergometer, respectively. The recovery time constant of oxygen consumption (day 1 bands and ergometer: 23.2 ± 3.7 s, 27.6 ± 6.5 s, respectively; day 2 bands and ergometer: 25.5 ± 5.4 s, 25.0 ± 4.9 s, respectively) had mean CVs of 10% and 11% for bands and ergometer, respectively. We conclude that measurements of oxygen consumption and mitochondrial capacity using NIRS can be obtained with good reproducibility.

Reduced skeletal muscle oxidative capacity and impaired training adaptations in heart failure

Systolic heart failure (HF) is associated with exercise intolerance that has been attributed, in part, to skeletal muscle dysfunction. The purpose of this study was to compare skeletal muscle oxidative capacity and training‐induced changes in oxidative capacity in participants with and without HF. Participants with HF (n = 16, 65 ± 6.6 years) were compared with control participants without HF (n = 23, 61 ± 5.0 years). A subset of participants (HF: n = 7, controls: n = 5) performed 4 weeks of wrist‐flexor exercise training. Skeletal muscle oxidative capacity was determined from the recovery kinetics of muscle oxygen consumption measured by near‐infrared spectroscopy (NIRS) following a brief bout of wrist‐flexor exercise. Oxidative capacity, prior to exercise training, was significantly lower in the HF participants in both the dominant (1.31 ± 0.30 min−1 vs. 1.59 ± 0.25 min−1, P = 0.002; HF and control groups, respectively) and nondominant arms (1.29 ± 0.24 min−1 vs. 1.46 ± 0.23 min−1, P = 0.04; HF and control groups, respectively). Following 4 weeks of endurance training, there was a significant difference in the training response between HF and controls, as the difference in oxidative training adaptations was 0.69 ± 0.12 min−1 (P < 0.001, 95% CI 0.43, 0.96). The wrist‐flexor training induced a ~50% improvement in oxidative capacity in participants without HF (mean difference from baseline = 0.66 ± 0.09 min−1, P < 0.001, 95% CI 0.33, 0.98), whereas participants with HF showed no improvement in oxidative capacity (mean difference from baseline = −0.04 ± 0.08 min−1, P = 0.66, 95% CI −0.24, 0.31), suggesting impairments in mitochondrial biogenesis. In conclusion, participants with HF had reduced oxidative capacity and impaired oxidative adaptations to endurance exercise compared to controls.

Skeletal muscle metabolic adaptations to endurance exercise training are attainable in mice with simvastatin treatment.

We tested the hypothesis that a 6-week regimen of simvastatin would attenuate skeletal muscle adaptation to low-intensity exercise. Male C57BL/6J wildtype mice were subjected to 6-weeks of voluntary wheel running or normal cage activities with or without simvastatin treatment (20 mg/kg/d, n = 7–8 per group). Adaptations in in vivo fatigue resistance were determined by a treadmill running test, and by ankle plantarflexor contractile assessment. The tibialis anterior, gastrocnemius, and plantaris muscles were evaluated for exercised-induced mitochondrial adaptations (i.e., biogenesis, function, autophagy). There was no difference in weekly wheel running distance between control and simvastatin-treated mice (P = 0.51). Trained mice had greater treadmill running distance (296%, P<0.001), and ankle plantarflexor contractile fatigue resistance (9%, P<0.05) compared to sedentary mice, independent of simvastatin treatment. At the cellular level, trained mice had greater mitochondrial biogenesis (e.g., ~2-fold greater PGC1α expression, P<0.05) and mitochondrial content (e.g., 25% greater citrate synthase activity, P<0.05), independent of simvastatin treatment. Mitochondrial autophagy-related protein contents were greater in trained mice (e.g., 40% greater Bnip3, P<0.05), independent of simvastatin treatment. However, Drp1, a marker of mitochondrial fission, was less in simvastatin treated mice, independent of exercise training, and there was a significant interaction between training and statin treatment (P<0.022) for LC3-II protein content, a marker of autophagy flux. These data indicate that whole body and skeletal muscle adaptations to endurance exercise training are attainable with simvastatin treatment, but simvastatin may have side effects on muscle mitochondrial maintenance via autophagy, which could have long-term implications on muscle health.

Measuring reactive hyperemia in the lower limb using near-infrared spectroscopy

Abstract. Near-infrared spectroscopy (NIRS) has been used to measure reactive hyperemia following a vascular occlusion. However, the procedures and methods of analysis used have varied. The purpose of the present study is to identify reproducible methods for measuring reactive hyperemia using HbO2 NIRS signals in the calf and foot. Healthy participants (10 male, 10 female) aged 19 to 28 years performed one of two tests: reproducibility trials or elevation protocol (30 and 60 cm limb elevation above the heart). The time to 50% reperfusion (T1/2) and the second (R2q) quartile rates of reperfusion were found to be the most reproducible parameters (coefficient of variation=7.12 to 14.1%). The time to 95% reperfusion (T95) was 12.7% more reproducible on average than the previously reported parameter of time to peak hyperemia. Measures of reperfusion time and rate slowed with increasing limb elevation. Correlations were identified between the calf and foot in the measurements of R2q (R2=0.713, p=0.021), T1/2 (R2=0.673, p=0.033), and T95 (R2=0.792, p=0.006). Half and 95% recovery times and second and third quartile rates expressed good reproducibility and sensitivity to change with reduced perfusion pressure. NIRS measures of reactive hyperemia have the potential to evaluate microvascular perfusion in clinical populations.

Comparisons of Ultrasound‐estimated Intramuscular Fat with Fitness and Health Indicators

We examined ultrasound‐estimated intramuscular fat of 4 muscles: rectus femoris (RF), biceps femoris (BF), tibialis anterior (TA), and medial gastrocnemius (MG), and compared the results with other health measures.

Two-photon deep-tissue spatially resolved mitochondrial imaging using membrane potential fluorescence fluctuations

Cell metabolism and viability are directly reflected in their mitochondria. Imaging-based analysis of mitochondrial morphological structure, size and dynamic characteristics can therefore provide critical insight into cell function. However, mitochondria are often very abundant, and due to their close to diffraction-limit size, it is often non-trivial to distinguish a tubular or large mitochondrion from an ensemble of punctate mitochondria. In this paper, we use membrane potential dependent fluorescence fluctuations of individual mitochondria to resolve them using an approach similar to single molecule localization microscopy. We use 2-photon microscopy to image mitochondrial intensity fluctuations at 200 μm deep inside an intact in-vivo mouse soleus muscle. By analyzing the acquired images, we can reconstruct images with an extra layer of information about individual mitochondria, separated from their ensemble. Our analysis shows a factor of 14 improvement in detection of mitochondria.