1. The role of the myoplasmic free Ca2+ concentration ([Ca2+]1) in the reduction of muscle force following contractions with stretch was investigated in single fibres from mouse toe muscle.Muscle fibres were either stretched by 25% of their optimum length (Lo) for ten tetani (Protocol I) or stretched by 50% of L. for between ten and thirty tetani (Protocol II). Hough (1902) recognized the condition of exercise-induced muscle damage which followed prolonged and unaccustomed exercise and was characterized by muscle pain, tenderness and weakness which took a number of days to recover.

We have examined the extent to which prolonged reductions in low-frequency force (i.e., low-frequency fatigue) result from increases in intracellular free Ca2+ concentration ([Ca2+]i) and alterations in muscle metabolites. Force and [Ca2+]i were measured in mammalian single muscle fibers in response to short, intermediate, and long series of tetani that elevated the [Ca2+]i-time integral to 5, 17, and 29 microM x s, respectively. Only the intermediate and long series resulted in prolonged (>60 x min) reductions in Ca2+ release and low-frequency fatigue. When fibers recovered from the long series of tetani without glucose, Ca2+ release was reduced to a greater extent and force was reduced at high and low frequencies. These findings indicate that the decrease in sarcoplasmic reticulum Ca2+ release associated with fatigue has at least two components: 1) a metabolic component, which, in the presence of glucose, recovers within 1 h, and 2) a component dependent on the elevation of the [Ca2+]i-time integral, which recovers more slowly. It is this Ca2+-dependent component that is primarily responsible for low-frequency fatigue.

1. The effect of muscle length on the myoplasmic free Ca2+ concentration ([Ca2+]1) and tetanic force production of intact single muscle fibres was investigated in the mouse flexor brevis muscle. Muscle fibres were analysed at 100 ,um intervals from 100 ,um shorter than the optimum length (Lo) to 300 ,um longer than Lo.

Single fibres were dissected from mouse flexor digitorum brevis muscles and subjected to a protocol of eccentric stretches consisting of ten tetani each with a 40 % stretch. Ten minutes later the fibres showed a reduced force, a shift in the peak of the force-length relation and a steepening of the force-frequency relation. Addition of the fluorescent dye sulforhodamine B to the extracellular space enabled the T-tubular system to be visualized. In unstimulated fibres and fibres subjected to 10 isometric tetani, the T-tubules were clearly delineated. Sulforhodamine B diffused out of the T-tubules with a half-time of 18 ± 1 s. Following the eccentric protocol, vacuoles connected to the T-tubules were detected in six out of seven fibres. Sulforhodamine B diffused out of the vacuoles of eccentrically damaged fibres extremely slowly with a half-time of 6.3 ± 2.4 min and diffused out of the T-tubules with a half-time of 39 ± 4 s. Vacuole production was eliminated by application of 1 m ouabain to the muscle during the eccentric protocol. On removal of the ouabain, vacuoles appeared over a period of 1 h and were more numerous and more widely distributed than in the absence of ouabain. We propose that T-tubules are liable to rupture during eccentric contraction probably because of the relative movement associated with the inhomogeneity of sarcomere lengths. Such rupture raises intracellular sodium and when the sodium is pumped from the cell by the sodium pump, the volume load of Na + and water exceeds the capacity of the T-tubules and causes vacuole production. The damage to the T-tubules may underlie a number of the functional changes that occur in eccentrically damaged muscle fibres.

Eccentric muscle contractions generate delayed onset muscle soreness (DOMS), possibly as a result of the high tensions involved causing muscle damage. Muscle function, serum indicators of muscle damage, and DOMS were investigated throughout a training regimen that involved a 40-min eccentric walk down a 25% gradient on a treadmill at 6.4 km/h once a week for 8 wk. Serum creatine kinase and myoglobin concentrations were used as indicators of muscle damage, and both demonstrated a delayed increase after the exercise protocol. The muscles that contracted eccentrically exhibited low-frequency fatigue, as well as decreases in muscle fatigability and maximal voluntary contraction force, which were greatest immediately postexercise. Although the results show that training reduces DOMS, the serum muscle protein response, and muscle function impairment, the time courses of these adaptations are different. It is suggested that the function of the muscle can be impaired without apparent muscle damage.