Department of Neuroregeneration, Netherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and Sciences, Amsterdam, The Netherlands.
PLoS One. 2013 May 28;8(5):e63862. doi: 10.1371/journal.pone.0063862. Print 2013.
Multiple genetic and environmental factors play a role in the development and progression of Parkinson's disease (PD). The main neuropathological hallmark of PD is the degeneration of dopaminergic (DAergic) neurons in the substantia nigra pars compacta. To study genetic and molecular contributors to the disease process, there is a great need for readily accessible cells with prominent DAergic features that can be used for reproducible in vitro cellular screening. Here, we investigated the molecular phenotype of retinoic acid (RA) differentiated SH-SY5Y cells using genome wide transcriptional profiling combined with gene ontology, transcription factor and molecular pathway analysis. We demonstrated that RA induces a general neuronal differentiation program in SH-SY5Y cells and that these cells develop a predominantly mature DAergic-like neurotransmitter phenotype. This phenotype is characterized by increased dopamine levels together with a substantial suppression of other neurotransmitter phenotypes, such as those for noradrenaline, acetylcholine, glutamate, serotonin and histamine. In addition, we show that RA differentiated SH-SY5Y cells express the dopamine and noradrenalin neurotransmitter transporters that are responsible for uptake of MPP(+), a well known DAergic cell toxicant. MPP(+) treatment alters mitochondrial activity according to its proposed cytotoxic effect in DAergic neurons. Taken together, RA differentiated SH-SY5Y cells have a DAergic-like phenotype, and provide a good cellular screening tool to find novel genes or compounds that affect cytotoxic processes that are associated with PD.
多种遗传和环境因素在帕金森病 (PD) 的发展和进展中起作用。PD 的主要神经病理学标志是黑质致密部多巴胺能 (DAergic) 神经元的退化。为了研究疾病过程中的遗传和分子贡献者，非常需要具有明显 DAergic 特征的易于获得的细胞，这些细胞可用于可重复的体外细胞筛选。在这里，我们使用全基因组转录谱结合基因本体、转录因子和分子途径分析，研究了视黄酸 (RA) 分化的 SH-SY5Y 细胞的分子表型。我们证明 RA 诱导 SH-SY5Y 细胞中的一般神经元分化程序，并且这些细胞发育出主要的成熟 DAergic 样神经递质表型。这种表型的特征是多巴胺水平增加，同时其他神经递质表型（如去甲肾上腺素、乙酰胆碱、谷氨酸、血清素和组氨酸）受到显著抑制。此外，我们表明 RA 分化的 SH-SY5Y 细胞表达多巴胺和去甲肾上腺素神经递质转运体，这些转运体负责摄取 MPP(+)，MPP(+) 是一种已知的 DAergic 细胞毒物。根据其在 DAergic 神经元中的拟毒作用，MPP(+) 处理改变线粒体活性。总之，RA 分化的 SH-SY5Y 细胞具有 DAergic 样表型，为寻找影响与 PD 相关的细胞毒性过程的新基因或化合物提供了良好的细胞筛选工具。
