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Parkin mutation decreases neurite complexity and maturation in neurons derived from human fibroblasts.
Brain Research Bulletin ( IF 3.8 ) Pub Date : 2020-03-07 , DOI: 10.1016/j.brainresbull.2020.03.006
Jiali Pu 1 , Ting Gao 1 , Ran Zheng 1 , Yi Fang 1 , Yang Ruan 1 , Chongyao Jin 1 , Ting Shen 1 , Jun Tian 1 , Baorong Zhang 1
Affiliation  

BACKGROUND Parkinson's disease (PD) is one of the most common neurodegenerative disorders, and mainly characterized by the progressive degeneration of dopaminergic (DA) neurons in the midbrain substantia nigra and non-DA neurons in many other parts of the brain. Previous studies have shown that several genes associated with the causes of PD can influence neurite outgrowth. Mutations of PRKN (encoding parkin, an E3 ubiquitin ligase) are the most frequent cause of recessively inherited PD. The lack of a PD phenotype in Prkn-knockout mice may imply a unique vulnerability of neurons to parkin mutations. METHODS CRISPR/Cas9 technology was used to target random mutations into exon3 of PRKN in human fibroblasts cell line MRC-5. The induced DA neurons were achieved from direct conversion of fibroblasts (with or without PRKN mutations) via a cocktail of transcriptional factors (Ascl1, Nurr1, Lmx1a, miRNA124, p53 shRNA) and chemicals (CHIR99021, Purmorphamine, TGFβ3, BDNF, GDNF, NGF and Y27632). RESULTS Herein, we successfully established human neuronal cell models with parkin mutations from fibroblast-reprogrammed neurons. In these neurons, not only were the induced ratio and number of mature neurons markedly decreased, but also the complexity of the neuronal processes, measured by total neurite length and number of terminals, was greatly reduced, in TH+ and TH-neurons with PRKN mutations. CONCLUSIONS The results suggest that parkin not only maintains the morphological complexity of human neurons, but also influences maturation and differentiation in the fibroblast reprogramming process.

中文翻译:

Parkin 突变降低了源自人类成纤维细胞的神经元的神经突复杂性和成熟度。

背景帕金森病(PD)是最常见的神经退行性疾病之一,其主要特征在于中脑黑质中的多巴胺能(DA)神经元和大脑的许多其他部分中的非DA神经元的进行性退化。先前的研究表明,与 PD 原因相关的几个基因会影响神经突的生长。PRKN(编码 parkin,一种 E3 泛素连接酶)的突变是隐性遗传 PD 的最常见原因。Prkn 基因敲除小鼠缺乏 PD 表型可能意味着神经元对 parkin 突变具有独特的脆弱性。方法 CRISPR/Cas9 技术用于靶向随机突变到人成纤维细胞 MRC-5 中 PRKN 的外显子 3。诱导的 DA 神经元是通过转录因子(Ascl1、Nurr1、Lmx1a、miRNA124、p53 shRNA)和化学物质(CHIR99021、Purmorphamine、TGFβ3、BDNF、GDNF、NGF和 Y27632)。结果在此,我们成功地建立了具有来自成纤维细胞重编程神经元的parkin 突变的人类神经元细胞模型。在这些神经元中,在具有 PRKN 突变的 TH+ 和 TH-神经元中,不仅成熟神经元的诱导比例和数量显着降低,而且通过总神经突长度和末端数量测量的神经元过程的复杂性也大大降低. 结论 结果表明,parkin 不仅保持了人类神经元的形态复杂性,
更新日期:2020-03-09
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