This study evaluated the effect of short bouts of stretching on the soleus muscle after immobilization, by measuring the birefringence of the intramuscular connective tissue (ICT) and the muscle fiber area. Thirty rats were divided into five groups: the left soleus was immobilized in the shortened position; after immobilization the animals remained free; after immobilization, the soleus was stretched daily (10 stretches of 60 sec followed by 30 sec rest); after immobilization, the soleus was stretched 3 times a week; control. Immobilization caused a loss of birefringence of the ICT and of muscle fiber area and only daily stretching increased both compared with the control (p< 0.01). In conclusion, short daily bouts of stretching after immobilization induced molecular reorganization of the collagen bundles and muscle fiber hypertrophy in the rat soleus.

In order to gain insight into intracellular mechanisms involved in longitudinal growth of skeletal muscle, we determined gene expression of ubiquitin-ligases (MAFbx/atrogin-1, E3 alpha, and MuRF-1) and deubiquitinating enzymes (UBP45, UBP69, and USP28) at different time-points (24, 48, and 96 h) of continuous stretch of the soleus and tibialis anterior (TA) muscles. In the soleus, real-time polymerase chain reaction (PCR) showed that MAFbx/atrogin-1, E3 alpha, and MuRF-1 gene expression was downregulated, peaking at 24-48 h. Gene expression of all deubiquitinating enzymes increased with continuous stretch of soleus. In the TA, gene expression of the ubiquitin-ligases MAFbx/atrogin-1 and MuRF-1 was elevated, whereas expression of UBP45 and UBP69 was downregulated. Western blot analysis showed that the overall ubiquitination level decreased in the soleus and increased in the TA during stretch. These results suggest that ubiquitin-ligases and deubiquitinating enzymes are involved in longitudinal growth induced by continuous muscle stretch.