title: Absence of aquaporin-4 in skeletal muscle alters proteins involved in bioenergetic pathways and calcium handling.
abstract: Aquaporin-4 (AQP4) is a water channel expressed at the sarcolemma of fast-twitch skeletal muscle fibers, whose expression is altered in several forms of muscular dystrophies. However, little is known concerning the physiological role of AQP4 in skeletal muscle and its functional and structural interaction with skeletal muscle proteome. Using AQP4-null mice, we analyzed the effect of the absence of AQP4 on the morphology and protein composition of sarcolemma as well as on the whole skeletal muscle proteome. Immunofluorescence analysis showed that the absence of AQP4 did not perturb the expression and cellular localization of the dystrophin-glycoprotein complex proteins, aside from those belonging to the extracellular matrix, and no alteration was found in sarcolemma integrity by dye extravasation assay. With the use of a 2DE-approach (BN/SDS-PAGE), protein maps revealed that in quadriceps, out of 300 Coomassie-blue detected and matched spots, 19 proteins exhibited changed expression in AQP4(-/-) compared to WT mice. In particular, comparison of the protein profiles revealed 12 up- and 7 down-regulated protein spots in AQP4-/- muscle. Protein identification by MS revealed that the perturbed expression pattern belongs to proteins involved in energy metabolism (i.e. GAPDH, creatine kinase), as well as in Ca(2+) handling (i.e. parvalbumin, SERCA1). Western blot analysis, performed on some significantly changed proteins, validated the 2D results. Together these findings suggest AQP4 as a novel determinant in the regulation of skeletal muscle metabolism and better define the role of this water channel in skeletal muscle physiology.
authors.affiliation: Department of General and Environmental Physiology, University of Bari-Aldo Moro, Bari, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Basco
authors.forename: Grazia Paola
authors.initials: GP
authors.lastname: Nicchia
authors.forename: Angelo
authors.initials: A
authors.lastname: D'Alessandro
authors.forename: Lello
authors.initials: L
authors.lastname: Zolla
authors.forename: Maria
authors.initials: M
authors.lastname: Svelto
authors.forename: Antonio
authors.initials: A
authors.lastname: Frigeri
mesh_terms: D000818:Animals; D051401:Aquaporin 4 / Q000172:deficiency*; D002118:Calcium / Q000378:metabolism*; D016189:Dystrophin / Q000378:metabolism; D004734:Energy Metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D025941:Protein Interaction Mapping; D040901:Proteomics; D052097:Quadriceps Muscle / Q000166:cytology / Q000378:metabolism; D012508:Sarcolemma / Q000378:metabolism
chemical_list: D051401:Aquaporin 4; D016189:Dystrophin; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D002118:Calcium