title: Proteomic profiling of the spinal cord in ALS: decreased ATP5D levels suggest synaptic dysfunction in ALS pathogenesis.
abstract: We aimed to gain new insights into the pathogenesis of sporadic ALS (sALS) through a comprehensive proteomic analysis.
  
  
  
  Protein profiles of the anterior and posterior horn in post-mortem spinal cord samples of 10 ALS patients and 10 controls were analysed using 2D-differential gel electrophoresis. The identified protein spots with statistically significant level changes and a spot ratio >2.0 were analysed by LC-MS/MS.
  
  
  
  In the posterior horn only 3 proteins were differentially expressed. In the anterior horn, 16 proteins with increased levels and 2 proteins with decreased levels were identified in ALS compared to controls. The identified proteins were involved in mitochondrial metabolism, calcium homeostasis, protein metabolism, glutathione homeostasis, protein transport and snRNP assembly. The two proteins with decreased levels, ATP5D and calmodulin, were validated by Western blot and immunostaining. Immunohistochemical and immunofluorescent double staining of ATP5D and synaptophysin showed that the reduction of ATP5D was most pronounced at synapses.
  
  
  
  We speculate that mitochondrial dysfunction in synaptic clefts could play an important role in sALS pathogenesis. A similar approach revealed decreased calmodulin expression mainly in the neuronal body and dendrites of ALS patients. These findings contribute to a deeper understanding of the disease process underlying ALS.
authors.affiliation: a Department of Neurology , Academic Medical Centre , Amsterdam , The Netherlands.
authors.forename: Jooyeon
authors.initials: J
authors.lastname: Engelen-Lee
authors.affiliation: b Department of Translational Neuroscience , Brain Centre Rudolf Magnus, University Medical Centre Utrecht , Utrecht , The Netherlands.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Blokhuis
authors.affiliation: c Department of Pathology , University Medical Centre Utrecht , Utrecht , The Netherlands.
authors.forename: Wim G M
authors.initials: WGM
authors.lastname: Spliet
authors.affiliation: b Department of Translational Neuroscience , Brain Centre Rudolf Magnus, University Medical Centre Utrecht , Utrecht , The Netherlands.
authors.forename: R Jeroen
authors.initials: RJ
authors.lastname: Pasterkamp
authors.affiliation: d Department of (Neuro)Pathology , Academic Medical Centre , Amsterdam , The Netherlands.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Aronica
authors.affiliation: e Proteomics Centre, Erasmus University Medical Centre , Rotterdam , The Netherlands.
authors.forename: Jeroen A A
authors.initials: JAA
authors.lastname: Demmers
authors.affiliation: c Department of Pathology , University Medical Centre Utrecht , Utrecht , The Netherlands.
authors.forename: Roel
authors.initials: R
authors.lastname: Broekhuizen
authors.affiliation: f Department of Molecular Biochemistry and Pharmacology , Mario Negri Institute for Pharmacological Research , Milano , Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Nardo
authors.affiliation: c Department of Pathology , University Medical Centre Utrecht , Utrecht , The Netherlands.|g Laboratory of Translational Immunology , University Medical Centre Utrecht , Utrecht , The Netherlands , and.
authors.forename: Niels
authors.initials: N
authors.lastname: Bovenschen
authors.affiliation: h Department of Neurology and Neurosurgery , Brain Centre Rudolf Magnus, University Medical Centre Utrecht , Utrecht , The Netherlands.
authors.forename: Leonard H
authors.initials: LH
authors.lastname: Van Den Berg
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000473:pathology*; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000096:biosynthesis / Q000235:genetics; D015536:Down-Regulation / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D020869:Gene Expression Profiling; D006706:Homeostasis / Q000235:genetics; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D008928:Mitochondria / Q000473:pathology; D025261:Mitochondrial Proton-Translocating ATPases; D011506:Proteins / Q000378:metabolism; D040901:Proteomics; D006180:Proton-Translocating ATPases / Q000235:genetics*; D013116:Spinal Cord / Q000737:chemistry / Q000473:pathology*; D013569:Synapses / Q000473:pathology*
chemical_list: D002147:Calmodulin; D011506:Proteins; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases; C579804:ATP5F1D protein, human; D002118:Calcium
keywords: ATP5D; Amyotrophic lateral sclerosis; Two-Dimensional Differential Gel Electrophoresis (2D-DIGE); proteomics; spinal cord; synaptic mitochondrial defect