...
首页> 外文期刊>Journal of neuroendocrinology >Selective Roles of Protein Kinase C Isoforms on Cell Motility of GT1 Immortalized Hypothalamic Neurones.
【24h】

Selective Roles of Protein Kinase C Isoforms on Cell Motility of GT1 Immortalized Hypothalamic Neurones.

机译:蛋白激酶C同工型对GT1永生的下丘脑神经元的细胞运动的选择性作用。

获取原文
获取原文并翻译 | 示例
           

摘要

Recently, we demonstrated that activation of the protein kinase C (PKC) signalling pathway promoted morphological differentiation of GT1 hypothalamic neurones via an increase in beta-catenin, a cell-cell adhesion molecule, indicating a possible involvement of PKC in cellular motility. In this study, we explored the differential roles of PKC isoforms in GT1 cell migration. First, we transiently transfected GT1 cells with enhanced green fluorescence protein (EGFP)-tagged actin to monitor the dynamic rearrangement of filamentous-actin (F-actin) in living cells. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a PKC activator, markedly promoted lamellipodia formation, while safingol (a PKCalpha-selective inhibitor) blocked the TPA-induced lamellipodial actin structure. Both wound-healing and Boyden migration assays showed that TPA treatment promoted neuronal migration of GT1 cells; however, cotreatment of TPA with safingol or rottlerin (a PKCdelta-selective inhibitor) clearly blocked this TPA effect, indicating that both PKCalpha and PKCdelta may be positive regulators of neuronal migration. By contrast, PKCgamma-EGFP-expressing GT1 cells exhibited decreased cellular motility and weak staining for actin stress fibres, suggesting that PKCgamma may act as a negative mediator of cell migration in these neurones. Among the PKC downstream signal molecules, p130Cas, a mediator of cell migration, and its kinase, focal adhesion kinase (FAK), increased following TPA treatment; phosphorylation of p130Cas was induced in a PKCalpha-dependent manner. Together, these results demonstrate that PKCalpha promotes GT1 neuronal migration by activating focal adhesion complex proteins such as p130Cas and FAK.
机译:最近,我们证明了激活的蛋白激酶C(PKC)信号通路通过增加β-catenin(一种细胞间粘附分子)的方式促进了GT1下丘脑神经元的形态分化,表明PKC可能参与了细胞运动。在这项研究中,我们探索了PKC亚型在GT1细胞迁移中的不同作用。首先,我们用增强的绿色荧光蛋白(EGFP)标记的肌动蛋白瞬时转染了GT1细胞,以监测活细胞中丝状肌动蛋白(F-actin)的动态重排。用PKC活化剂12-O-十四烷酰phorbol-13-乙酸盐(TPA)处理可显着促进层状脂蛋白形成,而沙丁酚(PKCalpha选择性抑制剂)则可阻断TPA诱导的层状脂蛋白肌动蛋白结构。伤口愈合和博伊登迁移试验均表明,TPA处理可促进GT1细胞的神经元迁移。然而,TPA与safingol或rottlerin(一种PKCdelta选择性抑制剂)的共同治疗明显阻断了该TPA的作用,表明PKCalpha和PKCdelta可能都是神经元迁移的正调节剂。相比之下,表达PKCgamma-EGFP的GT1细胞表现出降低的细胞运动性和肌动蛋白应激纤维的弱染色,表明PKCgamma可能在这些神经元中充当细胞迁移的负调节剂。在TPA处理后,在PKC下游信号分子中,细胞迁移的介体p130Cas及其激酶粘着斑激酶(FAK)增加。以PKCalpha依赖性方式诱导p130Cas的磷酸化。总之,这些结果表明,PKCalpha通过激活诸如p130Cas和FAK的粘着斑复合蛋白来促进GT1神经元的迁移。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号