Division of Cell Biology and Physiology, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
J Neurosci Res. 2013 Jul;91(7):963-70. doi: 10.1002/jnr.23241. Epub 2013 May 7.
SH-SY5Y, control, and Parkinson's disease (PD) cybrids prepared from an Indian population were differentiated using retinoic acid (RA) for understanding their dopaminergic characteristics and neuritogenesis. Undifferentiated control and PD cybrids exhibited higher levels of TH mRNA, but lower c-RET expression, short neurites, low neuritic density, and low proportion of cells with neurites compared with the undifferentiated parent cell line, SH-SY5Y. The expression levels of DAT and Ptx3 were similar to SH-SY5Y. PD cybrids showed poor viability and lower differentiating potency than SH-SY5Y or control cybrids. RA treatment for 6 days elevated c-RET expression and corrected the neuritic morphology of the control, but not of PD cybrids. Cell viability was found to be reduced in differentiated control and PD cybrids. TH expression level was significantly elevated in SH-SY5Y following RA treatment, but not in both the cybrids. In differentiated control and PD cybrids, the TH immunofluorescence intensity was significantly lower compared with SH-SY5Y cells. MitoTracker Green fluorescence intensity of the mitochondria was higher in differentiated PD cybrids. Dopamine released into the medium was unaffected in the differentiated SH-SY5Y or in the control cybrids but was significantly elevated in PD cybrids. These results suggest that PD cybrids, differentiated or undifferentiated, maintained morphological and biochemical phenotypes significantly different from those of the control cybrids, or the differentiated SH-SY5Y cells, and therefore could be an ideal cellular model of the disease for pharmacological screening of drugs and for investigation of the pathophysiology of PD.
SH-SY5Y、对照和源自印度人群的帕金森病(PD)杂种细胞系用维甲酸(RA)诱导分化,以了解其多巴胺能特征和神经突生成。未分化的对照和 PD 杂种细胞系表现出更高水平的 TH mRNA,但 c-RET 表达较低,神经突较短,神经突密度较低,有神经突的细胞比例较低,与未分化的亲本细胞系 SH-SY5Y 相比。DAT 和 Ptx3 的表达水平与 SH-SY5Y 相似。PD 杂种细胞系的活力较差,分化能力低于 SH-SY5Y 或对照杂种细胞系。RA 处理 6 天可提高 c-RET 表达并纠正对照杂种细胞系的神经突形态,但不能纠正 PD 杂种细胞系的神经突形态。发现分化的对照和 PD 杂种细胞系的细胞活力降低。RA 处理后,SH-SY5Y 的 TH 表达水平显著升高,但在两种杂种细胞系中均未升高。在分化的对照和 PD 杂种细胞系中,TH 免疫荧光强度明显低于 SH-SY5Y 细胞。分化的 PD 杂种细胞系中线粒体的 MitoTracker Green 荧光强度较高。分化的 SH-SY5Y 或对照杂种细胞系中多巴胺释放到培养基中的量不受影响,但 PD 杂种细胞系中的多巴胺释放量显著升高。这些结果表明,PD 杂种细胞系,无论分化或未分化,均保持与对照杂种细胞系或分化的 SH-SY5Y 细胞显著不同的形态和生化表型,因此可能是用于药物筛选和研究 PD 病理生理学的理想疾病细胞模型。