Stępkowski Tomasz Maciej, Wasyk Iwona, Grzelak Agnieszka, Kruszewski Marcin
Centre for Radiobiology and Biological Dosimetry, Institute of Nuclear Chemistry and Technology, Dorodna 16, 03-195, Warsaw, Poland.
Department of Molecular Biophysics, University of Łódź, Banacha 12/16, 90-237, Łódź, Poland.
Cell Mol Neurobiol. 2015 Nov;35(8):1137-47. doi: 10.1007/s10571-015-0207-5. Epub 2015 May 19.
LUHMES cells, a recently established line of immortalized embryonic mesencephalic cells, are the novel in vitro model for studying Parkinson's disease (PD) and dopaminergic neuron biology. Phosphoglyceromutase 5 (PGAM5) is a mitochondrial protein involved in mitophagy, mitochondria dynamics, and other processes important for PD pathogenesis. We tested the impact of lentiviral overexpression of PGAM5 protein in LUHMES cells on their differentiation and expression of 84 PD-related genes. LUHMES cells were transduced with PGAM5 or mock and treated with 100 μM 6-hydroxydopamine (6-OHDA), a model PD neurotoxin. Real-Time PCR analysis revealed that the treatment with 6-OHDA-induced changes in expression of 44 PD-related genes. PGAM5 transduction alone did not cause alternations in PD-related genes expression, nor it affected changes in gene expression mediated by 6-OHDA. The 6-OHDA-induced PD-related gene expression profile of LUHMES cells is presented for the first time and widely discussed.
LUHMES细胞是最近建立的永生化胚胎中脑细胞系,是研究帕金森病(PD)和多巴胺能神经元生物学的新型体外模型。磷酸甘油酸变位酶5(PGAM5)是一种线粒体蛋白,参与线粒体自噬、线粒体动力学以及其他对PD发病机制很重要的过程。我们测试了慢病毒过表达PGAM5蛋白对LUHMES细胞分化和84个PD相关基因表达的影响。用PGAM5或空载体转导LUHMES细胞,并用模型PD神经毒素100μM 6-羟基多巴胺(6-OHDA)处理。实时PCR分析显示,用6-OHDA处理可诱导44个PD相关基因表达发生变化。单独的PGAM5转导不会引起PD相关基因表达的改变,也不会影响由6-OHDA介导的基因表达变化。首次展示并广泛讨论了6-OHDA诱导的LUHMES细胞的PD相关基因表达谱。