Department of Biological Science, Sookmyung Womenos University, Seoul, Korea.
BMB Rep. 2010 May;43(5):369-74. doi: 10.5483/bmbrep.2010.43.5.369.
The PC12 is the widely used cell line to study neuronal differentiation. We had extensively investigated the details of protein expression in differentiated PC12 cells by proteomic analysis. The cells were incubated at the presence of nerve growth factor. We had analyzed the expression changes in the differentiating PC12 cells by 2-dimensional electrophoresis and the identification of the proteins using MALDI-TOF MS. By comparing expression pattern in the time course, we identified the candidate genes which are associated with neuronal differentiation. Among these genes, we performed real-time PCR analysis to validate Idh3alpha expression by the time course. To identify the function of Idh3alpha in neuronal differentiation stage, the transfection of Idh3alpha to PC12 cells was performed. As a result, we proved that up-regulation of Idh3alpha causes reduction in neural differentiation of PC12 cells. Based on these data, we suggest that Idh3alpha plays a role to the neuronal differentiation.
PC12 细胞是广泛用于研究神经元分化的细胞系。我们通过蛋白质组学分析,深入研究了分化 PC12 细胞中蛋白质表达的细节。细胞在神经生长因子的存在下孵育。我们通过二维电泳分析了分化 PC12 细胞中的表达变化,并使用 MALDI-TOF MS 鉴定了蛋白质。通过比较时程中的表达模式,我们鉴定了与神经元分化相关的候选基因。在这些基因中,我们通过实时 PCR 分析验证了 Idh3alpha 表达的时程。为了鉴定 Idh3alpha 在神经元分化阶段的功能,将 Idh3alpha 转染至 PC12 细胞。结果表明,Idh3alpha 的上调导致 PC12 细胞的神经分化减少。基于这些数据,我们认为 Idh3alpha 在神经元分化中起作用。