title: [Detection the mutated protein aggregation and mitochondrial function in fibroblasts from amyotrophic lateral sclerosis patients with SOD1 gene mutations].
abstract: OBJECTIVE
  To explore mutant superoxide dismutase (SOD)1 protein expression and mitochondrial function in amyotrophic lateral sclerosis (ALS) patients' fibroblasts carrying different SOD1 mutations.
  
  
  METHODS
  SOD1 gene mutation was detected using PCR and direct sequencing. Skin fibroblasts of three familial ALS patients with mutations and age/gender matched controls obtained by a punch skin biopsy were cultured. We performed immunofluorescence staining and quantitative detection of SOD1 proteins and mitochondrial membrane potential. Also, we detected the intracellular ROS by flow cytometry.
  
  
  RESULTS
  We found that fibroblasts from familial ALS patients carried SOD1-V14M, SOD1-G16A, SOD1-C111Y mutation, respectively. The cytoplasm abnormal SOD1 protein aggregates appeared in ALS patients carrying SOD1 mutations. And the cytoplasmic/nuclear ratio of SOD1 aggregates increased 2.54, 2.80, 3.25-fold for each mutations, respectively, compared to the control group. Three SOD1 mutant groups showed loss of mitochondrial membrane potential and the ratio of red / green fluorescence intensity decreased by 36%, 124%, 142%, respectively, compared to the control group. The intracellular ROS levels also increased 3.33, 3.65, and 6.87-fold respectively.
  
  
  CONCLUSIONS
  This work highlights that ALS alters SOD1 protein expression, mitochondrial function, and increases the ROS level even in peripheral tissues outside the central nervous system. Fibroblasts might therefore represent a powerful and minimally invasive tool to investigate ALS pathogenic mechanisms, which might translate into considerable advances in clinical management of the disease.
authors.affiliation: Weifang Medical University, Weifang 261053, China.
authors.forename: W C
authors.initials: WC
authors.lastname: Liu
authors.forename: T
authors.initials: T
authors.lastname: Liu
authors.forename: Z H
authors.initials: ZH
authors.lastname: Liu
authors.forename: M
authors.initials: M
authors.lastname: Deng
mesh_terms: D000690:Amyotrophic Lateral Sclerosis; D005347:Fibroblasts; D006801:Humans; D008928:Mitochondria; D009154:Mutation; D000072105:Superoxide Dismutase-1
chemical_list: C000606290:SOD1 protein, human; D000072105:Superoxide Dismutase-1