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Calpain/calpastatin activities and substrate depletion patterns during hindlimb unweighting and reweighting in skeletal muscle.

Enns D.L., Raastad T., Ugelstad I., Belcastro A.N.

Unloading of skeletal muscle by hindlimb unweighting (HU) is characterized by atrophy, protein loss, and an elevation in intracellular Ca(2+) levels that may be sufficient to activate Ca(2+)-dependent proteases (calpains). In this study, we investigated the time course of calpain activation and the depletion pattern of a specific structural protein (desmin) with unloading and subsequent reweighting. Rats underwent 12 h, 24 h, 72 h or 9 days of HU, followed by reweighting for either 0, 12 or 24 h. Total calpain-like activity was elevated with HU in skeletal muscle (P < 0.05) and was further enhanced with reweighting (P < 0.05). The increases in calpain-like activity were associated with a proportional increase in activity of the particulate fraction (P < 0.05). Activity of the mu-calpain isoform was elevated with 12 and 24 h of HU (P < 0.05) and returned to control levels thereafter. With reweighting, activities of mu-calpain were elevated above control levels for all HU groups except 9 days (P < 0.05). In contrast, minimal changes in m-calpain and calpastatin activity were observed with HU and reweighting. Although desmin depletion levels did not reach statistical significance, a significant inverse relationship was found between the mu-calpain/calpastatin ratio and the amount of desmin in isolated myofibrils (R = -0.83, P < 0.001). The results suggest that calpain activation is an early event during unloading in skeletal muscle, and that the majority of the increase in calpain activity can be attributed to the micro-isoform.

Eur. J. Appl. Physiol. 100:445-455(2007) [PubMed] [Europe PMC]

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