Scanning electron microscopy of tenotomized soleus muscles of the rat

The effects of tenotomy on the surface morphology of muscle fibers including myotendinous junctions in the rat soleus muscle were studied by scanning electron microscopy (SEM). Using potassium hydroxide (KOH) and collagenase, the extracellular materials were successfully removed to expose the surface of muscle fibers. When the soleus muscle was tenotomized at both proximal and distal ends, virtually all muscle fibers showed marked alterations of the fiber surface characterized by the formation of numerous transverse grooves and folds along their length. Narrow longitudinal grooves and folds of the sarcolemma were also observed. At myotendinous junctions, the fiber ends showed an over-all rounded shape with several short sarcoplasmic processes, indicating that the processes were significantly retracted. These changes were clearly recognizable at 5 days after tenotomy, and most apparent at one week. Thin-section electron microscopy of the same SEM samples demonstrated that such folding of the sarcolemma was not directly related to the sarcomere pattern of the underlying myofibrils, suggesting that, once formed, the folds and grooves were retained for a certain period of time. At 2 and 3 weeks the surface morphology of the fibers underwent a recovery process of restoring the smooth surface on which the cross-striations of the underlying myofibrils were seen. At the fiber ends, sarcoplasmic processes regrew into slender, wavy and short forms. Such sarcoplasmic processes were greater in number and more elaborate than those in the control muscle. At 5 and 6 weeks the fiber surface resumed an almost normal morphology, except that the sarcoplasmic processes at the fiber end were still shorter and more numerous than those in the control. These observations support our previous results obtained by thin-section electron microscopy that the myotendinous junction undergoes a series of morphological changes of collapse and regrowth of the sarcoplasmic processes, reflecting changes in the underlying myofibrils. In conclusion, the changes in the surface morphology of tenotomized muscle fibers were well correlated chronologically to those of myofibrils such as the central core lesion.