Gu Jun, Bao Yifeng, Chen Jian, Huang Chuanjun, Zhang Xinghua, Jiang Rui, Liu Qianqian, Liu Yonghua, Xu Xide, Shi Wei
Department of Neurosurgery, Affiliated Hospital of Nantong UniversityNantong, China; Department of Neurosurgery, Yancheng Third People's HospitalYancheng, China.
Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University Nantong, China.
Front Cell Neurosci. 2016 Dec 22;10:282. doi: 10.3389/fncel.2016.00282. eCollection 2016.
The proliferation and differentiation of neural stem cells (NSCs) is important for neural regeneration after cerebral injury. Here, for the first time, we show that phosphorylated (p)-ser847-nNOS (NP847), rather than nNOS, may play a major role in NSC proliferation after traumatic brain injury (TBI). Western blot results demonstrated that the expression of NP847 and Sox2 in the hippocampus is up-regulated after TBI, and they both peak 3 days after brain injury. In addition, an immunofluorescence experiment indicated that NP847 and Sox2 partly co-localize in the nuclei of NSCs after TBI. Further immunoprecipitation experiments found that NP847 and Sox2 can directly interact with each other in NSCs. Moreover, in an OGD model of NSCs, NP847 expression is decreased, which is followed by the down-regulation of Sox2. Interestingly, in this study, we did not observe changes in the expression of nNOS in the OGD model. Further research data suggest that the NP847-Sox2 complex may play a major role in NSCs through the Shh/Gli signaling pathway in a CaMKII-dependent manner after brain injury.
神经干细胞(NSCs)的增殖和分化对脑损伤后的神经再生至关重要。在此,我们首次表明,磷酸化(p)-丝氨酸847-nNOS(NP847)而非nNOS,可能在创伤性脑损伤(TBI)后NSCs的增殖中起主要作用。蛋白质免疫印迹结果显示,TBI后海马中NP847和Sox2的表达上调,且在脑损伤后3天均达到峰值。此外,免疫荧光实验表明,TBI后NP847和Sox2部分共定位于NSCs的细胞核中。进一步的免疫沉淀实验发现,NP847和Sox2在NSCs中可直接相互作用。此外,在NSCs的氧糖剥夺(OGD)模型中,NP847表达降低,随后Sox2表达下调。有趣的是,在本研究中,我们未观察到OGD模型中nNOS表达的变化。进一步的研究数据表明,脑损伤后,NP847-Sox2复合物可能通过CaMKII依赖性方式经Shh/Gli信号通路在NSCs中起主要作用。