Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
J Neurosci Res. 2011 Jul;89(7):1070-8. doi: 10.1002/jnr.22625. Epub 2011 Apr 6.
The use of immortalized cells has been instrumental as a tool with which to study gene regulation. However, it is crucial to understand the status of a given cell line, especially when investigating the regulation of genes whose expression is developmentally regulated. Several immortalized cell lines have been derived from primary cultures of mouse oligodendrocytes. Two such cell lines, N20.1 and Oli-neu, were characterized here in terms of their relative expression of myelin genes at both the mRNA level and the protein level. Analysis of the splice isoforms expressed by the myelin proteolipid protein (Plp1), myelin basic protein (Mbp), and 2',3'-cyclic nucleotide 3'-phosphodiesterase (Cnp) genes, along with the relative amount of protein expressed by these genes, suggests that the cell lines are representative of immature oligodendrocytes, although Oli-neu cells appear to be farther along the differentiation pathway compared with N20.1 cells. Previous studies have shown that the developmental increase in Plp1 gene expression that occurs during the active myelination period is governed by transcription regulatory elements present within the first intron. The responsiveness of one of these elements, the so-called antisilencer/enhancer (ASE), was investigated in both cell lines. Results presented here suggest that the ASE has a much more potent effect in Oli-neu cells. Thus, the two cell lines appear to be at different stages and will be useful as a means to study transcription regulatory elements whose influence changes during development.
永生化细胞的应用作为研究基因调控的工具是非常重要的。然而，了解给定细胞系的状态至关重要，尤其是在研究其表达受发育调控的基因的调节时。已经从小鼠少突胶质细胞的原代培养物中衍生出了几种永生化细胞系。在这里，我们对两种这样的细胞系，N20.1 和 Oli-neu，在 mRNA 水平和蛋白质水平上相对表达髓鞘基因的情况进行了特征描述。对髓鞘蛋白脂蛋白（Plp1）、髓鞘碱性蛋白（Mbp）和 2'，3'-环核苷酸 3'-磷酸二酯酶（Cnp）基因表达的剪接异构体进行分析，以及这些基因表达的蛋白质的相对量表明，这些细胞系代表不成熟的少突胶质细胞，尽管与 N20.1 细胞相比，Oli-neu 细胞似乎更接近分化途径。先前的研究表明，在活跃的髓鞘形成期间发生的 Plp1 基因表达的发育性增加受第一内含子中存在的转录调节元件控制。在这两种细胞系中研究了这些元件之一，即所谓的反沉默子/增强子（ASE）的反应性。这里提出的结果表明，ASE 在 Oli-neu 细胞中具有更强的作用。因此，这两种细胞系似乎处于不同的阶段，将作为研究其影响在发育过程中发生变化的转录调节元件的有用手段。
