Slit2 inactivates GSK3β to signal neurite outgrowth inhibition.
Slit2 inactivates GSK3β to signal neurite outgrowth inhibition.
复制标题
DOI：
10.1371/journal.pone.0051895
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhou FQ
中科院分区：
文献类型：
--
作者：
Byun J;Kim BT;Kim YT;Jiao Z;Hur EM;Zhou FQ
狭缝分子包括轴突引导线索的四个典型家族之一，这些家族在发育中的神经系统中引导生长锥。除了它们在轴突寻路中的作用外，新出现的证据表明，广泛的细胞过程受到Slit的调节，范围从神经突生长期间的分支形成和成束到肿瘤进展和血管生成。然而，Slit下游的分子和细胞机制在很大程度上仍然是未知的，部分原因是缺乏一个容易操纵的系统，产生容易识别的性状，以响应Slit。本研究证明了使用细胞系CAD作为分析系统来剖析由Slit触发的信号通路的可行性。在这里，我们表明，CAD细胞表达受体的狭缝（Robo1和Robo2）和CAD细胞响应纳摩尔浓度的Slit2显着减速的速度过程的延伸。使用该系统，我们揭示了Slit2使GSK 3 β失活，并且Slit2需要抑制GSK 3 β来抑制过程生长。此外，我们发现Slit2诱导GSK3β磷酸化并抑制成年背根神经节神经元中的轴突生长，从而验证了Slit2信号在原代神经元中的作用。考虑到CAD细胞可以使用标准分子生物学方法方便地操作，并且由Slit 2调节的过程延伸表型可以容易地追踪和定量，使用细胞系CAD将有助于鉴定下游效应子和阐明由Slit触发的信号级联。
Slit molecules comprise one of the four canonical families of axon guidance cues that steer the growth cone in the developing nervous system. Apart from their role in axon pathfinding, emerging lines of evidence suggest that a wide range of cellular processes are regulated by Slit, ranging from branch formation and fasciculation during neurite outgrowth to tumor progression and to angiogenesis. However, the molecular and cellular mechanisms downstream of Slit remain largely unknown, in part, because of a lack of a readily manipulatable system that produces easily identifiable traits in response to Slit. The present study demonstrates the feasibility of using the cell line CAD as an assay system to dissect the signaling pathways triggered by Slit. Here, we show that CAD cells express receptors for Slit (Robo1 and Robo2) and that CAD cells respond to nanomolar concentrations of Slit2 by markedly decelerating the rate of process extension. Using this system, we reveal that Slit2 inactivates GSK3β and that inhibition of GSK3β is required for Slit2 to inhibit process outgrowth. Furthermore, we show that Slit2 induces GSK3β phosphorylation and inhibits neurite outgrowth in adult dorsal root ganglion neurons, validating Slit2 signaling in primary neurons. Given that CAD cells can be conveniently manipulated using standard molecular biological methods and that the process extension phenotype regulated by Slit2 can be readily traced and quantified, the use of a cell line CAD will facilitate the identification of downstream effectors and elucidation of signaling cascade triggered by Slit.
登录
查看更多内容
影响因子：
64.5
作者：
Brose, K;Bland, KS;Kidd, T
通讯作者：
Kidd, T
影响因子：
34.7
作者：
Hur, Eun-Mi;Zhou, Feng-Quan
通讯作者：
Zhou, Feng-Quan
影响因子：
8
作者：
Dallol, A;Krex, D;Latif, F
通讯作者：
Latif, F
影响因子：
10.5
作者：
Hur, Eun-Mi;Saijilafu;Zhou, Feng-Quan
通讯作者：
Zhou, Feng-Quan
影响因子：
10.5
作者：
Lundström, A;Gallio, M;Samakovlis, C
通讯作者：
Samakovlis, C