Department of Neurology, University of California, Davis, School of Medicine, Sacramento, California, USA.
J Neurosci Res. 2010 Apr;88(5):957-70. doi: 10.1002/jnr.22262.
NG2 chondroitin sulfate proteoglycan is a surface marker of oligodendroglial progenitor cells (OPCs) in various species. In contrast to well-studied rat OPCs, however, we found that purified mouse NG2 surface positive cells (NG2(+) cells) require additional activation of cyclic AMP (cAMP) signaling for survival in a medium containing 30% B104 neuroblastoma conditioned medium supplemented with fibroblast growth factor-2 (B104CM+FGF2), whereas B104CM+FGF2 alone is sufficient for survival and selective proliferation of rat OPCs. After induction of in vitro differentiation, more than 90% of mouse NG2(+) cells became O4-positive, and a majority expressed myelin basic protein by 5 day of differentiation, which confirmed the identity of isolated mouse NG2(+) cells as OPCs. In comparison to rat OPCs, mouse OPCs in B104CM+FGF2 were less motile, and demonstrated lower basal phosphorylation levels of ERK1/2 and cAMP response element-binding protein (CREB) and a higher incidence of apoptosis mediated by the intrinsic pathway. Transient up-regulation of cAMP-CREB signaling partially inhibited apoptosis of mouse OPCs independently of the ERK pathway. This study demonstrates a difference in trophic requirements between mouse and rat OPCs, with an essential role for cAMP signaling to preserve viability of mouse OPCs.
NG2 硫酸软骨素蛋白聚糖是各种物种少突胶质前体细胞(OPC)的表面标志物。然而,与研究充分的大鼠 OPC 不同,我们发现纯化的小鼠 NG2 表面阳性细胞(NG2(+)细胞)在含有 30% B104 神经母细胞瘤条件培养基补充成纤维细胞生长因子-2(B104CM+FGF2)的培养基中需要额外激活环腺苷酸(cAMP)信号才能存活,而 B104CM+FGF2 单独即可用于大鼠 OPC 的存活和选择性增殖。体外诱导分化后,超过 90%的小鼠 NG2(+)细胞成为 O4 阳性,并且在分化的第 5 天大多数表达髓鞘碱性蛋白,这证实了分离的小鼠 NG2(+)细胞作为 OPC 的身份。与大鼠 OPC 相比,B104CM+FGF2 中的小鼠 OPC 迁移能力较低,ERK1/2 和 cAMP 反应元件结合蛋白(CREB)的基础磷酸化水平较低,并且由内在途径介导的细胞凋亡发生率较高。cAMP-CREB 信号的短暂上调可部分抑制小鼠 OPC 的细胞凋亡,而不依赖于 ERK 途径。这项研究表明了小鼠和大鼠 OPC 之间营养需求的差异,cAMP 信号在维持小鼠 OPC 的生存能力方面起着重要作用。