1. Our purpose was to compare the susceptibilities of muscles in animals of different ages to the injuries induced by stretching the contracting muscle. Single stretches provide an effective method for studying the factors that contribute to the initiation of contraction-induced injury. We hypothesized that, for maximally activated muscles in old compared with young or adult mice, the work input during a single stretch of any given strain is not different, but for a given work input the magnitude of the injury is greater. 2. The force deficit resulting from each single stretch was calculated as the decrease in the maximum isometric force expressed as a percentage of the maximum force prior to the stretch. Force deficits were compared 1 min after single stretches of in situ extensor digitorum longus (EDL) muscles of young, adult and old mice. In addition, measurements of force deficits immediately following single stretches of single permeabilized fibre segments from EDL muscles of young and old rats permitted investigation of the initial injury at the level of the contractile apparatus. 3. For maximally activated EDL muscles in young, adult and old mice, no differences were observed for the work input during stretches of any given strain. Furthermore, the relationships between the work and the resultant force deficit were not different for muscles in young and adult mice. In contrast, compared with the work-force deficit relationships for muscles in either young or adult mice, the relationship was significantly steeper for muscles in old mice. For single permeabilized fibres from muscles of old rats, the force deficits immediately after single stretches were greater than those observed for fibres from muscles of young rats. We conclude that the increased susceptibility of muscles in old animals to contraction-induced injury resides at least in part within the myofibrils.