Background: This study describes the ultrastructure of long-term denervated rat extensor digitorum longus and tibialis anterior muscles, with particular emphasis on understanding the cellular basis for the reduced restorative capacity of long-term denervated muscle. Methods: In 30 male WI/HicksCar rats, the right hindleg was denervated for periods of 1, 2, 4, 5.5, 6, 7, 12, 14, and 18 months before tissues were prepared for electron microscopy. Results: Atrophy of muscle fibers was prominent by the second month post-denervation. At this time, type II fibers showed greater atrophy than type I fibers. At further periods of denervation, atrophy of all fibers was seen; and with increasing times of denervation the muscle fibers became surrounded by dense mats of collagen fibers. Muscle spindles persisted for the duration of this study. At two and four months, satellite cells showed signs of activation, such as elongated cytoplasmic processes and an increased concentration of cytoplasmic organelles. As denervation progressed , activated satellite cells became more widely separated from their associated muscle fibers, and basal lamina material was deposited between the satellite cells and muscle fibers. Some satellite cells broke free from their muscle fibers, and others acted as bridges between two muscle fibers. Evidence was seen of both muscle fiber degeneration and the regeneration of new muscle fibers, often more than one regenerating fiber beneath a single basal lamina. Loose folds of basal lamina were often present around atrophic muscle fibers. As denervation progressed, the morphology of individual muscle fibers varied. Some contained well-ordered lattice arrays of myofilaments, whereas in others considerable sarcomeric disorganization was evident. Mitochondria became smaller and rounded; elements of the sarcoplasmic reticulum proliferated and became more disorganized; lipid droplets, glycogen deposits, and autopha-gic vesicles were all present in the cytoplasm of atrophic muscle fibers. Conclusions: In addition to muscle fiber atrophy, long-term denervated muscles show evidence of myofiber and capillary death, as well as the deposition of massive amounts of interstitial collagen. These changes, all of which would appear to reduce the restorative capacity of the dener-vated muscle, take place concurrently with the morphological activation of satellite cells. The latter indicates that even in the denervated condition , restorative processes occur concurrently with regressive processes.

Background In order to understand the cellular basis underlying the progressively poorer restorative capacity of long‐term denervated muscle, we determined the effects of long‐term denervation on the muscle fibers and satellite cell population of the rat extensor digitorum longus (EDL) muscle. Methods In 36 male rats, the right hind legs were denervated, and EDL muscles were removed 2, 4, 7, 12, and 18 months later. Muscles were either fixed for electron microscopic analysis or were dissociated into individual muscle fibers for direct fiber counting or for confocal microscopic analysis. Results The percentage of satellite cells rose from the 2.8% control value to 9.1% at 2 months of denervation; thereafter the percentage decreased to 1.1% at 18 months of denervation. The number of myonuclei per muscle fiber steadily declined from 410 in 4 month control muscle to 158 in 7 month denervated muscle. Up to 7 months of denervation, the total number of muscle fibers per muscle remained relatively constant at somewhat over 5,000. The calculated total satellite cell population in 4 month denervated EDL muscle was the same as that of controls at 65,000, but by 7 months of denervation it had declined to 21,000. With increasing time of denervation, the number of cross‐sectional profiles of muscle fibers not containing nuclei rose from 14% in control muscle to 49% in 12 month denervated muscle. This was correlated with a pronounced regular clumping of the nuclei, with pronounced nonnucleated segments between nuclear clumps. Conclusions Increasing times of denervation are accompanied by a pronounced decline in the number of myonuclei per muscle fiber and an initial rise and subsequent fall in satellite cell number. These changes are correlated with a decreasing restorative ability of these muscles over the same periods of denervation. Further work on the proliferative capacity of the remaining satellite cells is necessary before firm quantitative conclusions can be made. Anat. Rec. 248:346‐354 1997. © 1997 Wiley‐Liss, Inc.

Background: In order to understand the cellular basis underlying the progressively poorer restorative capacity of long-term denervated muscle, we determined the effects of long-term denervation on the muscle fibers and satellite cell population of the rat extensor digitorum longus (EDL) muscle.Methods: In 36 male rats, the right hind legs were denervated, and EDL muscles were removed 2, 4, 7, 12, and 18 months later. Muscles were either fixed for electron microscopic analysis or were dissociated into individual muscle fibers for direct fiber counting or for confocal microscopic analysis.Results: The percentage of satellite cells rose from the 2.8% control value to 9.1% at 2 months of denervation; thereafter the percentage decreased to 1.1% at 18 months of denervation. The number of myonuclei per muscle fiber steadily declined from 410 in 4 month control muscle to 158 in 7 month denervated muscle. Up to 7 months of denervation, the total number of muscle fibers per muscle remained relatively constant at somewhat over 5,000. The calculated total satellite cell population in 4 month denervated EDL muscle was the same as that of controls at 65,000, but by 7 months of denervation it had declined to 21,000. With increasing time of denervation, the number of cross-sectional profiles of muscle fibers not containing nuclei rose from 14% in control muscle to 49% in 12 month denervated muscle. This was correlated with a pronounced regular clumping of the nuclei, with pronounced nonnucleated segments between nuclear clumps.Conclusions: Increasing times of denervation are accompanied by a pronounced decline in the number of myonuclei per muscle fiber and an initial rise and subsequent fall in satellite cell number. These changes are correlated with a decreasing restorative ability of these muscles over the same periods of denervation. Further work on the proliferative capacity of the remaining satellite cells is necessary before firm quantitative conclusions can be made. Anat. Rec. 248:346-354 1997. r 1997 Wiley-Liss, Inc.

Background This study describes the ultrastructure of long‐term denervated rat extensor digitorum longus and tibialis anterior muscles, with particular emphasis on understanding the cellular basis for the reduced restorative capacity of long‐term denervated muscle. Methods In 30 male WI/HicksCar rats, the right hindleg was denervated for periods of 1, 2, 4, 5.5, 6, 7, 12, 14, and 18 months before tissues were prepared for electron microscopy. Results Atrophy of muscle fibers was prominent by the second month post‐denervation. At this time, type II fibers showed greater atrophy than type I fibers. At further periods of denervation, atrophy of all fibers was seen; and with increasing times of denervation the muscle fibers became surrounded by dense mats of collagen fibers. Muscle spindles persisted for the duration of this study. At two and four months, satellite cells showed signs of activation, such as elongated cytoplasmic processes and an increased concentration of cytoplasmic organelles. As denervation progressed, activated satellite cells became more widely separated from their associated muscle fibers, and basal lamina material was deposited between the satellite cells and muscle fibers. Some satellite cells broke free from their muscle fibers, and others acted as bridges between two muscle fibers. Evidence was seen of both muscle fiber degeneration and the regeneration of new muscle fibers, often more than one regenerating fiber beneath a single basal lamina. Loose folds of basal lamina were often present around atrophic muscle fibers. As denervation progressed, the morphology of individual muscle fibers varied. Some contained well‐ordered lattice arrays of myofilaments, whereas in others considerable sarcomeric disorganization was evident. Mitochondria became smaller and rounded; elements of the sarcoplasmic reticulum proliferated and became more disorganized; lipid droplets, glycogen deposits, and autophagic vesicles were all present in the cytoplasm of atrophic muscle fibers. Conclusions In addition to muscle fiber atrophy, long‐term denervated muscles show evidence of myofiber and capillary death, as well as the deposition of massive amounts of interstitial collagen. These changes, all of which would appear to reduce the restorative capacity of the denervated muscle, take place concurrently with the morphological activation of satellite cells. The latter indicates that even in the denervated condition, restorative processes occur concurrently with regressive processes. Anat. Rec. 248:355–365, 1997. © 1997 Wiley‐Liss, Inc.

The extensor digitorum longus muscle (EDL) or soleus muscle (SOL) in rats was mechanically overloaded on one side. The muscles were (i) untreated (normal) or (ii) self- or foreign-reinnervated (leading to persisting muscle fibres) or transplanted (leading to regenerating muscle fibres). The effects of the different procedures were studied in the treated and untreated muscles on the operated side and in the untreated muscles on the contralateral side. Overloading led to an absolute increase in mass (versus control values) in the normal muscles and to a relative increase in mass (versus the lower mass after reinnervation) in the treated muscles. The mechanism underlying this gain in mass was usually a compensatory hypertrophy. Overloading was followed by transformation of fibres from fast to slow in normal muscles. In the reinnervated muscles, the fibre distribution changed in response to the new nervous input and then remained constant. The majority of the experimental procedures elicited significant muscular changes in the contralateral muscles, including hyperplasia, fibre transformation and fibre hypertrophy or atrophy. The changes are interpreted as the consequence of a general compensatory neuromuscular activity designed to maintain a symmetrical posture during walking and running. These frequent and substantial muscular changes in the unoperated muscles clearly show that the muscles of the contralateral side cannot be used as normal controls.