Morphological alterations study in the Golgi apparatus of skin fibroblasts in spinocerebellar ataxia type 3 patients
10.3760/cma.j.cn113694-20211223-00915
- VernacularTitle:脊髓小脑共济失调3型患者皮肤成纤维细胞高尔基体变化的研究
- Author:
Yanlin WANG
1
;
Zhuoya WANG
;
Zhilei WANG
;
Huifang SUN
;
Qi ZHANG
;
Na CHEN
;
Yuming XU
Author Information
1. 郑州大学第一附属医院神经内科，郑州 450052
- Keywords:
Spinocerebellar ataxias;
Fibroblasts;
Skin;
Golgi apparatus;
Apoptosis;
Cell viability
- From:
Chinese Journal of Neurology
2022;55(9):993-1001
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To identify the morphological alterations in the Golgi apparatus of skin fibroblasts in spinocerebellar ataxia type 3 (SCA3) patients.Methods:In this study, 3 SCA3 patients and 3 healthy volunteers were obtained in the First Affiliated Hospital of Zhengzhou University from 2016 to 2020. The cytosine, adenine, and guanine repeats of 3 SCA3 patients were 14/76, 20/80 and 21/82, respectively. Tissue mass culture was used to amplify skin fibroblasts derived from SCA3 patients and healthy volunteers. Cell viability and apoptosis were detected using cell counting kit-8 assay and flow cytometry, respectively. Western blotting and immunofluorescence assay were used to detect the protein expression of ataxin-3, Golgi reassembly stacking protein 2 (GORASP2), and Golgi matrix protein 130 (GM130) in the skin fibroblasts. The morphology of the Golgi apparatus in skin fibroblasts was detected using transmission electron microscopy.Results:Tissue culture of skin fibroblasts of both SCA3 patients and healthy volunteers was successfully established. The patient-derived dermal fibroblasts expressing mutant ataxin-3 protein exhibited reduced cell viability ( t=5.06,P=0.007), increased apoptosis ( t=3.77, P=0.020), fragmentation of the Golgi apparatus, increased expression of GM130 ( t=5.23, P=0.006), and decreased expression of GORASP2 ( t=4.35, P=0.012). Transmission electron microscopy revealed that the Golgi apparatus was disorganized in skin fibroblasts. Conclusion:Fragmentation of the Golgi apparatus occurs in the skin fibroblasts of SCA3 patients, and abnormal morphology and structure of the Golgi apparatus may be involved in the pathogenesis of SCA3.
