Universiteit Hasselt, BIOMED, Agoralaan Building C, 3590 Diepenbeek, Belgium.
J Neurosci Methods. 2009 Oct 30;184(1):1-9. doi: 10.1016/j.jneumeth.2009.07.004. Epub 2009 Jul 10.
OLN-93 cells, a cell line established from spontaneously transformed rat brain glial cultures, are used as a model for oligodendrocytes. These cells are known to undergo morphological changes upon serum deprivation. The objective of the present study is to investigate a possible correlation between these morphological changes and (1) the loss or gain of oligodendrocyte markers and (2) the electrophysiological properties of these cells. Using RT-PCR and immunocytochemistry, we demonstrate that the OLN-93 cell line expresses a broad range of markers (NG2, CNP, MAG, MOG) both when cultured in medium containing 10% or 0.5% fetal calf serum. Whole-cell patch-clamp recordings demonstrate that, regardless of the culture conditions, OLN-93 cells mainly express delayed-rectifying K+ currents, a characteristic of immature oligodendrocytes. These currents are in part mediated by the shaker family of voltage-gated potassium channels. Kv1.1 and Kv1.3-expression are present at the mRNA and at the protein levels, and functional evidence for Kv1.3 mediated currents was obtained by using the selective blocker margatoxin. Under low serum conditions, OLN-93 cells exhibit differentiation-like morphological changes. However, we provide evidence that these morphological modifications do not necessarily correlate with biochemical or functional changes. Based on these data, we conclude that the OLN-93 cell line can be situated at a developmental stage between a late pre-oligodendrocyte and a late immature oligodendrocyte, regardless of serum concentration.
OLN-93 细胞是从自发转化的大鼠脑胶质培养物中建立的细胞系,用作少突胶质细胞的模型。已知这些细胞在血清剥夺时会发生形态变化。本研究的目的是研究这些形态变化与(1)少突胶质细胞标志物的丧失或获得,以及(2)这些细胞的电生理特性之间可能存在的相关性。通过 RT-PCR 和免疫细胞化学,我们证明 OLN-93 细胞系在含有 10%或 0.5%胎牛血清的培养基中培养时表达广泛的标志物(NG2、CNP、MAG、MOG)。全细胞膜片钳记录表明,无论培养条件如何,OLN-93 细胞主要表达延迟整流钾电流,这是未成熟少突胶质细胞的特征。这些电流部分由 shaker 家族电压门控钾通道介导。Kv1.1 和 Kv1.3 的表达存在于 mRNA 和蛋白水平,并且通过使用选择性阻断剂 margatoxin 获得了 Kv1.3 介导的电流的功能证据。在低血清条件下,OLN-93 细胞表现出分化样形态变化。然而,我们提供的证据表明,这些形态修饰不一定与生化或功能变化相关。基于这些数据,我们得出结论,无论血清浓度如何,OLN-93 细胞系都可以处于晚期前少突胶质细胞和晚期未成熟少突胶质细胞之间的发育阶段。