SERCA activation prevents Ca2+ and ATP upregulation during 3-day soleus muscle unloading in rats

Abstract

The imbalance in the ratio of protein synthesis versus protein degradation results in skeletal muscle atrophy following unloading. The onset of these processes is regulated by the sarcoplasmic concentrations of ATP and calcium (Ca²⁺). We tested the hypothesis that unloading-induced inactivation of sarcoendoplasmic reticulum calcium ATPase (SERCA) results in raised Ca²⁺ concentrations, triggering catabolic processes. CDN1163, an activator of SERCA, was used to test this hypothesis. Three groups of male rats were used: control rats with intraperitoneal injection of placebo (C), 3 days of unloading with placebo injection (3HS), and 3 days of unloading injected with CDN1163 (3HSC). Treatment with CDN1163 during 3 days of soleus muscle unloading prevented the upregulation of Ca²⁺ and ATP and the slow-to-fast shift in muscle fiber composition. This treatment blocked the decrease in the phosphorylation of the anabolic markers [glycogen synthase kinase-3β (GSK3β), eukaryotic translation elongation factor 2 (eEF2), and ribosomal protein S6 (S6, Ser240/244/Ser235/236)], and therefore it is likely that it improved the efficiency of translation in the unloaded muscle, but it did not affect mTORC1-dependent signaling. Treatment with CDN1163 also modulated the regulation of the Ca²⁺-dependent signaling in muscle during unloading via SERCA1 and calsequestrin 2 (CSQ2) and changes in the calcium/calmodulin-dependent protein kinase II (CaMKII) phosphorylation and the content of inositol 1,4,5-trisphosphate receptor (IP3R). In addition, CDN1163 prevented the upregulation of the mRNA expression of muscle-specific RING finger protein 1 (MuRF1) [but not ubiquitin ligase muscle atrophy F-box (MAFbx)] and attenuated the increase of casitas B lymphoma-b (Cbl-b) and ubiquitin mRNA expression during unloading. Activation of SERCA with CDN1163 prevents the upregulation of Ca²⁺ and ATP, as well as calcium-dependent and ubiquitin-proteasome pathways markers, and improves protein translation efficiency in 3-day unloaded soleus muscle.NEW & NOTEWORTHY A hypothesis was tested that unloading-induced inactivation of SERCA results in the accumulation of increased Ca²⁺ concentrations and activation of catabolic processes. CDN1163, an activator of SERCA, was used to test this hypothesis. CDN1163 prevented the decrease in phosphorylation of anabolic markers, which likely improved translation efficiency in unloaded muscle. CDN1163 prevented unloading-induced upregulation of mRNA expression of MuRF1 (but not MAFbx) and attenuated the increase of Cbl-b and ubiquitin mRNA expression.

Keywords: AMPK; ATP; CaMKII; atrophy; unloading.