Impact of aging on mitochondrial function in cardiac and skeletal muscle.

Both skeletal muscle and cardiac muscle are subject to marked structural and functional impairment with aging and these changes contribute to the reduced capacity for exercise as we age. Since mitochondria are involved in multiple aspects of cellular homeostasis including energetics, reactive oxygen species signaling, and regulation of intrinsic apoptotic pathways, defects in this organelle are frequently implicated in the deterioration of skeletal and cardiac muscle with aging. On this basis, the purpose of this review is to evaluate the evidence that aging causes dysfunction in mitochondria in striated muscle with a view towards drawing conclusions about the potential of these changes to contribute to the deterioration seen in striated muscle with aging. As will be shown, impairment in respiration and reactive oxygen species emission with aging are highly variable between studies and seem to be largely a consequence of physical inactivity. On the other hand, both skeletal and cardiac muscle mitochondria are more susceptible to permeability transition and this seems a likely cause of the increased recruitment of mitochondrial-mediated pathways of apoptosis seen in striated muscle. The review concludes by examining the role of degeneration of mitochondrial DNA versus impaired mitochondrial quality control mechanisms in the accumulation of mitochondria that are sensitized to permeability transition, whereby the latter mechanism is favored as the most likely cause.

Estimating the size of the capillary-to-fiber interface in skeletal muscle: a comparison of methods.

Current evidence suggests that the size of the capillary-to-fiber (C/F) interface is a major determinant of O2 flux into muscle fibers, and methods have been developed for estimating the size of this region via the C/F perimeter ratio in perfusion-fixed material (Mathieu-Costello O, Ellis CG, Potter RF, MacDonald IC, and Groom AC. Am J Physiol Heart Circ Physiol 261: H1617-H1625, 1991) and the quotient of the individual, fiber-based C/F number ratio and fiber perimeter (C/F perimeter exchange index) in muscle biopsies (Hepple RT. Can J Appl Physiol 22: 11-22, 1997). The purpose of this study was to compare the two methods and examine how differences in muscle tissue preparation between perfusion fixation and frozen biopsy can influence the estimate of the size of the C/F interface. The left medial gastrocnemius muscle of nine purpose-bred dogs was perfusion fixed in situ, and a sample from the midportion of the midbelly was processed for microscopy. A corresponding sample from the right gastrocnemius muscle obtained by open biopsy in six of the nine animals was frozen for histochemistry. A significant correlation was found between the two estimates of the size of the C/F interface in the same sections of perfusion-fixed material (r = 0.75, P < 0.05). However, estimates of the size of the C/F interface were smaller in biopsies than perfusion-fixed material, and there was no significant relationship between the estimates in the two preparations. This was due to differences in fiber size (33% larger fiber cross-sectional area in biopsy material after normalization for sarcomere length; P < 0.05) and muscle sampling between the two tissue preparations.

Structural basis of muscle O(2) diffusing capacity: evidence from muscle function in situ.

Although evidence for muscle O(2) diffusion limitation of maximal O(2) uptake has been found in the intact organism and isolated muscle, its relationship to diffusion distance has not been examined. Thus we studied six sets of three purpose-bred littermate dogs (aged 10-12 mo), with 1 dog per litter allocated to each of three groups: control (C), exercise trained for 8 wk (T), or left leg immobilized for 3 wk (I). The left gastrocnemius muscle from each animal was surgically isolated, pump-perfused, and electrically stimulated to peak O(2) uptake at three randomly applied levels of arterial oxygenation [normoxia, arterial PO(2) (Pa(O(2))) 77 +/- 2 (SE) Torr; moderate hypoxia, Pa(O(2)): 33 +/- 1 Torr; and severe hypoxia, Pa(O(2)): 22 +/- 1 Torr]. O(2) delivery (ml. min(-1). 100 g(-1)) was kept constant among groups for each level of oxygenation, with O(2) delivery decreasing with decreasing Pa(O(2)). O(2) extraction (%) was lower in I than T or C for each condition, but calculated muscle O(2) diffusing capacity (Dmus(O(2))) per 100 grams of muscle was not different among groups. After the experiment, the muscle was perfusion fixed in situ, and a sample from the midbelly was processed for microscopy. Immobilized muscle showed a 45% reduction of muscle fiber cross-sectional area (P < 0.05), and a resulting 59% increase in capillary density (P < 0.05) but minimal reduction in capillary-to-fiber ratio (not significant). In contrast, capillarity was not significantly different in T vs. C muscle. The results show that a dramatically increased capillary density (and reduced diffusion distance) after short-term immobilization does not improve Dmus(O(2)) in heavily working skeletal muscle.

Skeletal muscle: master or slave of the cardiovascular system?

Skeletal muscle and cardiovascular system responses to exercise are so closely entwined that it is often difficult to determine the effector from the affector. The purpose of this manuscript and its companion papers is to highlight (and perhaps assist in unraveling) the interdependency between skeletal muscle and the cardiovascular system in both chronic and acute exercise. Specifically, we elucidate four main areas: 1) how a finite cardiac output is allocated to a large and demanding mass of skeletal muscle, 2) whether maximal muscle oxygen uptake is determined peripherally or centrally, 3) whether blood flow or muscle metabolism set the kinetic response to the start of exercise, and 4) the matching of structural adaptations in muscle and the microcirculation in response to exercise. This manuscript, the product of an American College of Sports Medicine Symposium, unites the thoughts and findings of four researchers, each with different interests and perspectives, but with the common intent to better understand the interaction between oxygen supply and metabolic demand during exercise.

Skeletal muscle: microcirculatory adaptation to metabolic demand.

The issue of whether skeletal muscle is master or slave of the cardiovascular system depends on frame of reference. Acute manipulations of convective O2 delivery clearly show that O2 supply sets the upper limit of muscle VO2max. However, studies of adaptation to chronic conditions such as training and hypoxia show that skeletal muscle has a remarkable capacity to meet changes in metabolic demand. Moreover, there are several lines of evidence that these adaptations are essential to changes in VO2max. Studies show that with training, electrical stimulation, and chronic hypoxia, the ratio of capillary surface per fiber surface and fiber mitochondrial volume/fiber length is preserved, suggesting a primary regulated feature in skeletal muscle is matching the structural capacity for O2 flux to mitochondrial metabolic demand. Adaptations in both capillarity and mitochondrial respiratory capacity have also been shown to be important components in the adaptive increase in VO2max with training. Collectively, this evidence argues against skeletal muscle being simply a slave to the cardiovascular system.

Rapid force recovery in contracting skeletal muscle after brief ischemia is dependent on O(2) availability.

We tested the hypothesis that contracting skeletal muscle can rapidly restore force development during reperfusion after brief total ischemia and that this rapid recovery depends on O(2) availability and not an alternate factor related to blood flow. Isolated canine gastrocnemius muscle (n = 5) was stimulated to contract tetanically (isometric contraction elicited by 8 V, 0.2-ms duration, 200-ms trains, at 50-Hz stimulation) every 2 s until steady-state conditions of muscle blood flow (controlled by pump perfusion) and developed force were attained (3 min). While maintaining the same stimulation pattern, muscle blood flow was then reduced to zero (complete ischemia) for 2 min. Normal blood flow was then restored to the contracting muscle; however, two distinct conditions of oxygenation (at the same blood flow) were sequentially imposed: deoxygenated blood (30 s), blood with normal arterial O(2) content (30 s), a return to deoxygenated blood (30 s), and finally a return to normal arterial O(2) content (90 s). During the ischemic period, force development fell to 39 +/- 6 (SE)% of normal (from 460 +/- 40 to 170 +/- 20 N/100 g). When muscle blood flow was restored to normal by perfusion with deoxygenated blood, developed force continued to decline to 140 +/- 20 N/100 g. Muscle force rapidly recovered to 310 +/- 30 N/100 g (P < 0.05) during the 30 s in which the contracting muscle was perfused with oxygenated blood and then fell again to 180 +/- 30 N/100 g when perfused with blood with low PO(2). These findings demonstrate that contracting skeletal muscle has the capacity for rapid recovery of force development during reperfusion after a short period of complete ischemia and that this recovery depends on O(2) availability and not an alternate factor related to blood flow restoration.

Oxygen uptake kinetics during exercise in chronic heart failure: influence of peripheral vascular reserve.

Exercise performance in chronic heart failure is severely impaired, due in part to a peripherally mediated limitation. In addition to impaired maximal exercise capacity, the O(2) uptake (VO(2)) response during submaximal exercise may be affected, with a greater reliance on anaerobiosis leading to early fatigue. However, the response of VO(2) kinetics to submaximal exercise in chronic heart failure has not been studied extensively; in particular, the relationship between oxygen utilization and the peripheral response to exercise has not been studied. The present investigation examined the time-constant (tau, corresponding to 63% of the total response fitted from exercise onset) of the VO(2) kinetics on-response to submaximal exercise and its relationship to maximal peripheral blood flow in patients with chronic heart failure, and compared responses with those in healthy sedentary subjects. Subjects were 10 patients with chronic heart failure (NYHA class II/III). The mean age was 50+/-12 years, with a mean resting left ventricular ejection fraction of 25+/-9%. Controls were 10 age-matched healthy subjects. VO(2(max)) was first determined for all subjects. Repeated transitions from rest to exercise were performed on a cycle ergometer while measuring breath-by-breath responses of VO(2) at a fixed work rate of 50% of VO(2(max)) (heart failure patients and healthy controls) and at a work rate equivalent to the average in heart failure patients (65 W; healthy controls only). On a separate occasion, post-maximal ischaemic exercise calf blood flow was measured (strain-gauge plethysmography). Whereas heart failure subjects displayed a significantly prolonged VO(2) kinetics response at a similar absolute workload (i.e. 65 W), as indicated by a longer tau value (42 s, compared with 22 s in controls; P<0.01), there was no difference in tau at a similar relative work rate [50% of VO(2(max))]. In addition, heart failure subjects demonstrated a lower maximal calf blood flow (P<0.05) than control subjects. These results indicate that patients with heart failure have a prolonged VO(2) kinetics on-response compared with healthy subjects at a similar absolute work rate (i.e. 65 W), but not at a similar relative work rate [50% of VO(2(max))]. Thus, despite a reduced maximal calf blood flow response associated with heart failure, it does not appear that this contributes to an impairment of the submaximal exercise response beyond that explained by a reduced maximal exercise capacity [VO(2(max))].

Dissociation of peak vascular conductance and V(O2) max among highly trained athletes.

Previously, a strong relationship has been found between whole body maximal aerobic power (VO(2 max)) and peak vascular conductance in the calf muscle (J. L. Reading, J. M. Goodman, M. J. Plyley, J. S. Floras, P. P. Liu, P. R. McLaughlin, and R. J. Shephard. J. Appl. Physiol. 74: 567-573, 1993; P. G. Snell, W. H. Martin, J. C. Buckley, and C. G. Blomqvist. J. Appl. Physiol. 62: 606-610, 1987), suggesting a matching between maximal exercise capacity and peripheral vasodilatory reserve across a broad range of aerobic power. In contrast, long-term training could alter this relationship because of the unique demands for muscle blood flow and cardiac output imposed by different types of training. In particular, the high local blood flows but relatively low cardiac output demand imposed by the type of resistance training used by bodybuilders may cause a relatively greater development in peripheral vascular reserve than in aerobic power. To examine this possibility, we studied the relationship between treadmill VO(2 max) and vascular conductance in the calf by using strain-gauge plethysmography after maximal ischemic plantar flexion exercise in 8 healthy sedentary subjects (HS) and 28 athletes. The athletes were further divided into three groups: 10 elite middle-distance runners (ER), 11 power athletes (PA), and 7 bodybuilders (BB). We found that both BB and ER deviate from the previously demonstrated relationship between VO(2 max) and vascular conductance. Specifically, for a given vascular conductance, BB had a lower VO(2 max), whereas ER had a higher VO(2 max) than did HS and PA. We conclude that the relationship between peak vascular conductance and aerobic power is altered in BB and ER because of training-specific effects on central vs. peripheral cardiovascular adaptation to local skeletal muscle metabolic demand.

Increased capillarity in leg muscle of finches living at altitude.

An increased ratio of muscle capillary to fiber number (capillary/fiber number) at altitude has been found in only a few investigations. The highly aerobic pectoralis muscle of finches living at 4,000-m altitude (Leucosticte arctoa; A) was recently shown to have a larger capillary/fiber number and greater contribution of tortuosity and branching to total capillary length than sea-level finches (Carpodacus mexicanus; SL) of the same subfamily (O. Mathieu-Costello, P. J. Agey, L. Wu, J. M. Szewczak, and R. E. MacMillen. Respir. Physiol. 111: 189-199, 1998). To evaluate the role of muscle aerobic capacity on this trait, we examined the less-aerobic leg muscle (deep portion of anterior thigh) in the same birds. We found that, similar to pectoralis, the leg muscle in A finches had a greater capillary/fiber number (1.42 +/- 0.06) than that in SL finches (0.77 +/- 0.05; P < 0.01), but capillary tortuosity and branching were not different. As also found in pectoralis, the resulting larger capillary/fiber surface in A finches was proportional to a greater mitochondrial volume per micrometer of fiber length compared with that in SL finches. These observations, in conjunction with a trend to a greater (rather than smaller) fiber cross-sectional area in A than in SL finches (A: 484 +/- 42, SL: 390 +/- 26 micrometer2, both values at 2.5-micrometer sarcomere length; P = 0.093), support the notion that chronic hypoxia is also a condition in which capillary-to-fiber structure is organized to match the size of the muscle capillary-to-fiber interface to fiber mitochondrial volume rather than to minimize intercapillary O2 diffusion distances.

Resistance and aerobic training in older men: effects on VO2peak and the capillary supply to skeletal muscle.

Both aerobic training (AT) and resistance training (RT) may increase aerobic power (VO2peak) in the older population; however, the role of changes in the capillary supply in this response has not been evaluated. Twenty healthy men (age 65-74 yr) engaged in either 9 wk of lower body RT followed by 9 wk of AT on a cycle ergometer (RT-->AT group) or 18 wk of AT on a cycle ergometer (AT-->AT group). RT was performed three times per week and consisted of three sets of four exercises at 6-12 repetitions maximum. AT was performed three times per week for 30 min at 60-70% heart rate reserve. VO2peak was increased after both RT and AT (P < 0.05). Biopsies (vastus lateralis) revealed that the number of capillaries per fiber perimeter length was increased after both AT and RT (P < 0.05), paralleling the changes in VO2peak, whereas capillary density was increased only after AT (P < 0.01). These results, and the finding of a significant correlation between the change in capillary supply and VO2peak (r = 0.52), suggest the possibility that similar mechanisms may be involved in the increase of VO2peak after high-intensity RT and AT in the older population.

A new measurement of tissue capillarity: the capillary-to-fibre perimeter exchange index.

The surface area of contact between capillaries and muscle fibres has been suggested to be the site of greatest oxygen flux density in the movement of oxygen from the capillaries to the muscle fibres. A new measurement of tissue capillarity, designed specifically for use on non-perfusion fixed muscle tissue (i.e., that obtained via needle biopsy), is presented that describes the capillary supply from this perspective. The Capillary-to-Fibre Perimeter Exchange Index (the CFPE Index) is derived as the quotient of the individual capillary-to-fibre ratio (i.e., the capillary-to-fibre ratio calculated for each fibre individually) and the fibre perimeter. This method is suggested to provide a means of quantitating potential alterations in the capacity for oxygen flux (e.g., as may occur in response to a training intervention) and any carrier- or receptor-mediated aspect of blood-tissue exchange between the capillaries and muscle-fibres (e.g., insulin or glucose delivery).

Quantitating the capillary supply and the response to resistance training in older men.

Resistance training (RT) has been shown to increase aerobic power in older humans. To determine the effects of RT on the capillary supply in this population, nine older men (65-74 y) engaged in 9 weeks RT of the lower body. Following RT, peak O2 uptake (V.O2,peak) increased by 7% (P<.01). Needle biopsies (vastus lateralis muscle) revealed significant increases (mean +/- SE) in fibre area (3,874 +/- 314 microm2 to 4,778 +/- 309 microm2), fibre perimeter (P, 262 +/- 11 microm to 296 +/- 11 microm), capillary contacts (3.7 +/- .2 to 4.3 +/- .3) and the individual capillary-to-fibre ratio (C:Fi, 1. 33 +/- .32 to 1.61 +/- .37, P<.005). To evaluate the potential for blood-tissue exchange, both fibre area-based and perimeter-based measures of the capillary supply were compared. While the area-based measures were maintained, C:Fi/P was increased, consistent with an increase in the size of the fibre-capillary interface and thus, an increased potential for oxygen flux following RT. Of the measurements of capillary supply, V.O2,peak correlated best with C:Fi/P (r = 0.69, P<.005). These results indicate a significant increase in the capillary supply relative to the perimeter, but not the cross-sectional area, of the muscle fibres following RT in older men, and that C:Fi/P is strongly correlated to the V.O2,peak in this population.