title: A transcriptomic approach to search for novel phenotypic regulators in McArdle disease.
abstract: McArdle disease is caused by lack of glycogen phosphorylase (GP) activity in skeletal muscle. Patients experience exercise intolerance, presenting as early fatigue and contractures. In this study, we investigated the effects produced by a lack of GP on several genes and proteins of skeletal muscle in McArdle patients. Muscle tissue of 35 patients and 7 healthy controls were used to identify abnormalities in the patients' transcriptomic profile using low-density arrays. Gene expression was analyzed for the influence of variables such as sex and clinical severity. Differences in protein expression were studied by immunoblotting and 2D electrophoresis analysis, and protein complexes were examined by two-dimensional, blue native gel electrophoresis (BN-PAGE). A number of genes including those encoding acetyl-coA carboxylase beta, m-cadherin, calpain III, creatine kinase, glycogen synthase (GS), and sarcoplasmic reticulum calcium ATPase 1 (SERCA1), were found to be downregulated in patients. Specifically, compared to controls, GS and SERCA1 proteins were reduced by 50% and 75% respectively; also, unphosphorylated GS and SERCA1 were highly downregulated. On BN-PAGE analysis, GP was present with GS in two muscle protein complexes. Our findings revealed some issues that could be important in understanding the physiological consequences of McArdle disease: (i) SERCA1 downregulation in patients could result in impaired calcium transport in type II (fast-twitch) muscle fibers, leading to early fatigability during exercise tasks involving type II fibers (which mostly use glycolytic metabolism), i.e. isometric exercise, lifting weights or intense dynamic exercise (stair climbing, bicycling, walking at a very brisk pace), (ii) GP and GS were found together in two protein complexes, which suggests a new regulatory mechanism in the activity of these glycogen enzymes.
authors.affiliation: Departament de Patologia Mitocondrial i Neuromuscular, Hospital Universitari Vall d'Hebron, Institut de Recerca, Universitat Autónoma de Barcelona, Barcelona, Spain.
authors.forename: Gisela
authors.initials: G
authors.lastname: Nogales-Gadea
authors.forename: Inés
authors.initials: I
authors.lastname: Consuegra-García
authors.forename: Juan C
authors.initials: JC
authors.lastname: Rubio
authors.forename: Joaquin
authors.initials: J
authors.lastname: Arenas
authors.forename: Marc
authors.initials: M
authors.lastname: Cuadros
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Camara
authors.forename: Javier
authors.initials: J
authors.lastname: Torres-Torronteras
authors.forename: Carmen
authors.initials: C
authors.lastname: Fiuza-Luces
authors.forename: Alejandro
authors.initials: A
authors.lastname: Lucia
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Martín
authors.forename: Elena
authors.initials: E
authors.lastname: García-Arumí
authors.forename: Antoni L
authors.initials: AL
authors.lastname: Andreu
mesh_terms: D016022:Case-Control Studies; D005786:Gene Expression Regulation; D024981:Glycogen Phosphorylase / Q000032:analysis; D006012:Glycogen Storage Disease Type V / Q000235:genetics*; D006006:Glycogen Synthase / Q000032:analysis; D006801:Humans; D046912:Multiprotein Complexes / Q000502:physiology; D018656:Muscle Fibers, Fast-Twitch; D010641:Phenotype; D040901:Proteomics; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000235:genetics; D059467:Transcriptome
chemical_list: D046912:Multiprotein Complexes; D024981:Glycogen Phosphorylase; D006006:Glycogen Synthase; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases