To determine whether the regulation of calpain system is involved in non-hibernators and hibernators in disused condition, the soleus (SOL) and extensor digitorum longus (EDL) muscles were used for investigating the muscle mass, the ratio of muscle wet weight/body weight (MWW/BW), fiber-type distribution, fiber cross-sectional area (CSA), and the protein expression of MuRF1, calpain-1, calpain-2, calpastatin, desmin, troponin T, and troponin C in hindlimb unloading rats and hibernating Daurian ground squirrels. The muscle mass, MWW/BW, and fiber CSA were found significantly decreased in SOL and EDL of hindlimb unloading rats, but unchanged in hibernating ground squirrels. The MuRF1 expression was increased in both SOL and EDL of unloading rats, while it was only increased in SOL, but maintained in EDL of hibernating ground squirrels. The expression levels of calpain-1 and calpain-2 were increased in different degrees in unloaded SOL and EDL in rats, while they were maintained in EDL and even reduced in SOL of hibernating ground squirrels. Besides, the expression of calpastatin was decreased in unloaded rats, but increased in hibernating ground squirrels. The desmin expression was decreased in unloaded rats, but maintained in hibernating squirrels. Interestingly, the levels of troponin T and troponin C were decreased in both SOL and EDL of unloaded rats, but increased in hibernating ground squirrels with muscle-type specificity. In conclusion, differential calpain activation and substrate-selective degradation in slow and fast muscles are involved in the mechanisms of muscle atrophy of unloaded rats and remarkable ability of muscle maintenance of hibernating ground squirrels.

We examined the lateral gastrocnemius (LG), plantaris (PL), and extensor digitorum longus (EDL) muscles to determine whether differential activation of the calpain system is related to the degree of atrophy in these fast-twitch skeletal muscles during hibernation in Daurian ground squirrels ( Spermophilus dauricus). Results from morphological indices showed various degrees of atrophy in the order LG > PL > EDL. Furthermore, all three muscles underwent fast-to-slow fiber-type conversion in hibernation. In regard to the calpain system in the LG muscle, cytosolic Ca2+ increased significantly in hibernation, followed by recovery in posthibernation. Furthermore, calpastatin expression significantly decreased, and calpain 1 and 2 expression significantly increased, which may be responsible for the increased degradation of desmin during hibernation compared with that during summer activity. In the EDL muscle, Ca2+ overload was observed during interbout arousal, and calpastatin showed an increase during hibernation and interbout arousal, which could explain the increased levels of troponin T during both periods compared with levels during summer activity. These findings suggest that cytosolic Ca2+ overload and subsequent calpain 1 and 2 activation may be an important mechanism of LG muscle atrophy during hibernation. Cytosolic Ca2+ homeostasis and high expression of calpain inhibitor calpastatin during hibernation may also be an important mechanism for the EDL muscle to maintain muscle mass. Thus, the differential activation of the calpain system and selective degradation of downstream substrates may be involved in muscle atrophy of different fast-twitch muscles during hibernation. NEW & NOTEWORTHY We found that the extent of both muscle atrophy and calpain system activation differed in fast-twitch lateral gastrocnemius (LG), plantaris (PL), and extensor digitorum longus (EDL) skeletal muscles in hibernating Daurian ground squirrels, but similar hierarchies in the order of LG > PL > EDL. The differential activation of the calpain system and selective degradation of downstream substrates may be involved in muscle atrophy in different fast-twitch muscles during hibernation.