Analysis by two-dimensional Blue Native/SDS-PAGE of membrane protein alterations in rat soleus muscle after hindlimb unloading.
Muscle atrophy occurring in several pathophysiological conditions determines decreases in muscle protein synthesis, increases in the rate of proteolysis and changes in muscle fiber composition. To determine the effect of muscle atrophy induced by hindlimb unloading (HU) on membrane proteins from rat soleus, a proteomic approach based on two-dimensional Blue Native/SDS-PAGE was performed. Proteomic analysis of normal and HU soleus muscle demonstrates statistically significant changes in the relative level of 36 proteins. Among the proteins identified by mass spectrometry, most are involved in pathways associated with muscle fuel utilization, indicating a shift in metabolism from oxidative to glycolytic. Moreover, immunoblotting analysis revealed an increase in aquaporin-4 (AQP4) water channel and an alteration of proteins belonging to the dystrophin-glycoprotein complex (DGC). AQP4 and DGC are regulated in soleus muscle subjected to simulated microgravity in response to compensatory mechanisms induced by muscle atrophy, and they parallel the slow-to-fast twitch conversion that occurs in soleus fibers during HU. In conclusion, the alterations of soleus muscle membrane proteome may play a pivotal role in the mechanisms involved in disuse-induced muscle atrophy.
Basco
Davide
D
Department of General and Environmental Physiology and Centre of Excellence in Comparative Genomics (CEGBA), University of Bari, 70126, Bari, Italy.
Nicchia
Grazia Paola
GP
Desaphy
Jean-François
JF
Camerino
Diana Conte
DC
Frigeri
Antonio
A
Svelto
Maria
M
D000818:Animals; D003911:Dextrans / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism / Q000502:physiology; D019417:Hindlimb Suspension / Q000502:physiology*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D018832:Molecular Chaperones / Q000032:analysis / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D009133:Muscular Atrophy / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D020543:Proteome / Q000032:analysis; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar
D003911:Dextrans; D008565:Membrane Proteins; D018832:Molecular Chaperones; D020543:Proteome; C017080:blue dextran