Cast immobilization of hindlimb upregulates sarcolipin expression in atrophied skeletal muscles and increases thermogenesis in C57BL/6J mice

Abstract

Mechanical unloading impairs cytosolic calcium (Ca²⁺) homeostasis in skeletal muscles. In this study, we investigated whether sarco(endo)plasmic reticulum Ca²⁺-ATPase (SERCA) itself or one of the regulators of the Ca²⁺ SERCA pump, sarcolipin (SLN), is altered to deregulate Ca²⁺ homeostasis in cast immobilized, atrophied muscles. Hindlimb muscles of 8-wk-old male C57BL/6J mice were subjected to bilateral cast immobilization for 2 wk. Two-week-cast immobilization induced both body weight and skeletal muscle loss. Highly phosphorylated Ca²⁺/calmodulin-dependent protein kinase II in the atrophied muscles suggested that cytosolic Ca²⁺ concentration was elevated. Extremely high expression levels of SLN mRNA and protein were observed in the atrophied muscles. Upregulation of SLN at the transcriptional level was supported by low RCAN1 expression, which is a negative regulator of SLN. We treated C₂C₁₂ cells with dexamethasone to mimic muscle atrophy in vitro and showed a direct relationship between high SLN mRNA expression and low Ca²⁺ uptake by sarcoplasmic reticulum. Since SLN reportedly plays a role in nonshivering thermogenesis, we performed a cold tolerance test of the whole body. As a result, we found that mice with cast immobilization showed high cold tolerance, suggesting that cast immobilization promoted whole body thermogenesis. Although the activity level was decreased during cast immobilization without change in food intake, adipose tissue weights also decreased significantly after cast immobilization. Concomitantly, we conclude that cast immobilization of hindlimb increased thermogenesis in C57Bl/6J mice, probably via high expression of SLN.

Keywords: calcium; immobilization; nonshivering thermogenesis; sarcolipin.