Protein Turnover in Rat Skeletal Muscle During In Vitro Hypo-Osmotic Stress

Abstract

Hypo-osmolality influences tissue metabolism, but research on protein turnover in skeletal muscle is limited. The purpose of this investigation was to examine the effects of hypo-osmotic stress on protein turnover in rat skeletal muscle. We hypothesized increased protein synthesis and reduced degradation following hypo-osmotic exposure. EDL muscles (n=8/group) were incubated in iso-osmotic (290 Osm/kg) or hypo-osmotic (190 Osm/kg) modified medium 199 (95% O2, 5% CO2, pH 7.4, 30±2 °C) for 60 min, followed by 75 min incubations with L-U[14C]phenylalanine or cycloheximide to determine protein synthesis and degradation. Immunoblotting was performed to assess signalling pathways involved. Phenylalanine uptake and incorporation were increased by 199% and 169% respectively in HYPO from ISO (p < 0.05). This was supported by elevated phosphorylation of mTOR Ser2448 (+12.5%) and increased Thr389 phosphorylation on p70s6 kinase (+23.6%) (p < 0.05). Hypo-osmotic stress increased protein synthesis and potentially amino acid uptake. Future studies should examine the upstream mechanisms involved.