Pathogenesis Of Amyotrophic Lateral Sclerosis Patients With Different FUS Gene Mutations

Objectives The expression of FUS gene mutation protein,reactive oxygen level and mitochondrial function were explored for skin fibroblasts of Amyotrophic lateral sclerosis(ALS)patients with different FUS gene mutations.Methods 1 FUS gene mutation was detected using PCR and direct sequencing.2 Skin fibroblasts of three familial ALS patients with mutations and age/gender matched controls obtained by a punch skin biopsy were cultured.3 We performed immunofluorescence staining and quantitative detection of FUS proteins.4 Flow cytometry was used to detect reactive oxygen species(ROS),mitochondrial membrane potential levels.5 The cell phenotype of skin fibroblasts was observed from time to time by using the Incu Cyte system of living cell workstation for 72 h.Results 1 We found that fibroblasts from familial ALS patients carrying FUS-R524 W,FUS-P525 L mutation,there was an abnormal accumulation of FUS protein in the fibroblasts,which was 2.1 and 2.2 times higher in the mutation group than in the control group,2 The level of ROS in the mutant cells was 31.1 and 65.1 times higher than that in the control group.3 Flow cytometry detection of mutant gene cells and healthy control cells showed no statistically significant difference in mitochondrial membrane potential changes.Conclusions This work highlights that abnormal accumulation of FUS protein in the fibroblasts of ALS patients with FUS gene mutation,and the ROS level of fibroblasts in ALS patients was significantly higher than that in healthy controls.However,there was no obvious change in the mitochondrial membrane potential The peripheral tissue fibroblasts of ALS patients have a potential model of short-term and rapid diagnosis and prognosis,which is more beneficial to the future clinical application and accelerate the process of ALS research.Figure 11;Table 4;Reference 95...