Nucleotides and epidermal growth factor induce parallel cytoskeletal rearrangements and migration in cultured adult murine neural stem cells
Nucleotides and epidermal growth factor induce parallel cytoskeletal rearrangements and migration in cultured adult murine neural stem cells
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DOI：
10.1111/j.1748-1716.2010.02092.x
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发表时间：
2010-06-01
影响因子：
6.3
通讯作者：
Zimmermann, H.
中科院分区：
文献类型：
--
作者：
Grimm, I.;Ullsperger, S. N.;Zimmermann, H.
目的：成年人脑室下区（SVZ）含有神经干细胞，这些神经干细胞产生迁移到嗅球（OB）并分化为中间神经元的神经母细胞。控制神经发生途径中的基本功能的分子线索，如增殖，短距离和长距离迁移，功能整合和细胞存活知之甚少。我们以前已经表明，培养的成人神经干细胞表达了相当多的各种核苷酸受体和核苷酸和表皮生长因子（EGF）诱导收敛的细胞内信号传导途径，进行潜在的协同作用，在控制神经干细胞增殖和细胞存活。在这里，我们研究EGF和核苷酸ATP，ADP β S和UTP在神经干细胞migration.Methods的作用：神经干细胞从成年小鼠分离，并进行贴壁培养。用四甲基罗丹明异硫氰酸盐-鬼笔环肽标记F-肌动蛋白。处理图像以定量评价荧光标记。激动剂诱导的AKT和黏着斑激酶的磷酸化通过定量Western印迹法进行分析。使用48孔microchemotaxis chambers.Results：核苷酸和EGF诱导应力纤维的形成，增加皮质肌动蛋白细胞骨架和细胞扩散的激动剂依赖性细胞迁移进行了测定。这与AKT和黏着斑激酶的磷酸化增加有关。使用microchemotaxis室，我们证明了一个平行的细胞migration.Conclusion增加：我们的研究结果表明，核苷酸和EGF作为旁分泌或自分泌的信号物质可以是相关的结构和维持SVZ的细胞结构和神经母细胞迁移到OB的流。
Aim:The adult subventricular zone (SVZ) contains neural stem cells that generate neuroblasts migrating to the olfactory bulb (OB) and differentiating into interneurones. The molecular cues controlling essential functions within the neurogenesis pathway such as proliferation, short and long distance migration, functional integration and cell survival are poorly understood. We have previously shown that cultured adult neural stem cells express a considerable variety of nucleotide receptors and that nucleotides and epidermal growth factor (EGF) induce converging intracellular signalling pathways that carry potential for synergism in the control of neural stem cell proliferation and cell survival. Here we investigate the role of EGF and the nucleotides ATP, ADP beta S and UTP in neural stem cell migration.Methods:Neural stem cells were prepared from adult mice and subjected to adherent culture. Labelling of F-actin was performed with tetramethylrhodamine isothiocyanate-phalloidin. Images were processed for quantitative evaluation of fluorescence labelling. Agonist-induced phosphorylation of AKT and focal adhesion kinase was analysed by quantitative Western blotting. Agonist-dependent cell migration was assayed using 48-well microchemotaxis chambers.Results:Nucleotides and EGF induce the formation of stress fibres, an increase in the cortical actin cytoskeleton and in cell spreading. This is associated with increased phosphorylation of AKT and focal adhesion kinase. Using microchemotaxis chambers we demonstrate a parallel increase in cell migration.Conclusion:Our results suggest that nucleotides and EGF acting as paracrine or autocrine signalling substances can be of relevance for structuring and maintaining the cytoarchitecture of the SVZ and the stream of neuroblasts migrating to the OB.