Neurogenomics Laboratory, Multiple Sclerosis Center, Sheba Medical Center, Israel.
Neurogenomics Laboratory, Multiple Sclerosis Center, Sheba Medical Center, Israel.
Neuroscience. 2017 Nov 19;364:107-121. doi: 10.1016/j.neuroscience.2017.09.018. Epub 2017 Sep 19.
Spontaneous neural repair from endogenous neural stem cells' (NSCs) niches occurs in response to central nervous system (CNS) injuries to only a limited extent. Uncovering the mechanisms that control neural repair and can be further manipulated to promote NSCs toward oligodendrocyte progenitors cells (OPCs) and myelinating oligodendrocytes is a major objective. In the current study, we describe high-throughput transcriptional changes in adult mouse subventricular zone (SVZ)-NSCs during differentiation in vitro. In order to identify myelin-specific transcriptional regulators among large transcriptional changes associated with differentiation, we have focused on transcripts encoding transcription factors and regulators showing expression profile that is highly correlated with expression of myelin-encoding genes. We have revealed previously undescribed effect of Prickle1 and Nfe2l3 transcriptional regulators that are positively correlated with expression of myelin basic protein (MBP). Using Prickle1 and Nfe2l3 silencing and immunocytochemistry approaches, we demonstrated that silencing of Prickle1 dramatically decreases differentiation to NG2OPCs while Nfe2l3 moderately decreases as compared with control siRNA. Moreover, silencing of Prickle1 also decreases maturation of OPCs to MBP oligodendrocytes (OLs). Accordingly, overexpression of Prickle1 increases the differentiation of NSCs to CNPase pre-myelinating and myelinating MBP OLs. In particular, the necessity of Prickle1 for oligodendrocyte differentiation is demonstrated in purified OPCs cultures. Our findings demonstrate the role of Prickle1 in positive regulation of differentiation and maturation of oligodendrocytes suggesting that targeting Prickle1 in CNS injuries and particularly in demyelinating disease could promote generation of myelinating oligodendrocytes from endogenous niches to replenish damaged oligodendrocytes.
内源性神经干细胞（NSCs）龛中的自发神经修复仅在中枢神经系统（CNS）损伤后在一定程度上发生。揭示控制神经修复的机制，并进一步操纵这些机制以促进 NSCs 向少突胶质前体细胞（OPCs）和髓鞘形成少突胶质细胞是主要目标。在本研究中，我们描述了成年小鼠侧脑室下区（SVZ）-NSC 在体外分化过程中的高通量转录变化。为了在与分化相关的大量转录变化中鉴定出髓鞘特异性转录调节剂，我们专注于编码转录因子和调节剂的转录本，这些转录本的表达谱与髓鞘编码基因的表达高度相关。我们揭示了 Prickle1 和 Nfe2l3 转录调节剂以前未描述的作用，它们与髓鞘碱性蛋白（MBP）的表达呈正相关。通过 Prickle1 和 Nfe2l3 沉默和免疫细胞化学方法，我们证明沉默 Prickle1 可显著减少向 NG2OPCs 的分化，而与对照 siRNA 相比，Nfe2l3 则适度减少。此外，沉默 Prickle1 还会减少 OPC 向 MBP 少突胶质细胞（OL）的成熟。因此，Prickle1 的过表达增加了 NSCs 向 CNPase 前髓鞘和髓鞘 MBP OL 的分化。特别是，在纯化的 OPC 培养物中证明了 Prickle1 对少突胶质细胞分化的必要性。我们的研究结果表明 Prickle1 在少突胶质细胞分化和成熟的正向调节中的作用，表明在 CNS 损伤，特别是在脱髓鞘疾病中靶向 Prickle1 可能会促进内源性龛位中髓鞘形成少突胶质细胞的产生，以补充受损的少突胶质细胞。
