Nerve growth factor induces neurite outgrowth of PC12 cells by promoting Gβγ-microtubule interaction.
Nerve growth factor induces neurite outgrowth of PC12 cells by promoting Gβγ-microtubule interaction.
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DOI：
10.1186/s12868-014-0132-4
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发表时间：
2014-12-31
期刊：
影响因子：
2.4
通讯作者：
Roychowdhury S
中科院分区：
文献类型：
--
作者：
Sierra-Fonseca JA;Najera O;Martinez-Jurado J;Walker EM;Varela-Ramirez A;Khan AM;Miranda M;Lamango NS;Roychowdhury S
微管的组装和拆卸对于神经突的生长和分化至关重要。有证据表明，神经生长因子（NGF）通过激活受体酪氨酸激酶TrkA诱导PC 12细胞的神经突起生长。G蛋白偶联受体（GPCR）以及异源三聚体G蛋白也参与调节神经突生长。然而，这些途径之间的可能联系，以及它们最终可能如何收敛到调节神经突生长过程中MT的组装和组织尚不清楚。在这里，我们报告，Gβγ，GPCR途径的重要组成部分，是神经生长因子诱导的PC 12细胞的神经元分化的关键。我们发现，NGF促进了Gβγ与MT的相互作用，刺激了MT的组装。虽然Gβγ螯合肽GRK 2 i抑制神经突形成，破坏MT，并诱导神经突损伤，但Gβγ激活剂mSIRK刺激神经突生长，这表明Gβγ参与了这一过程。因为我们之前已经表明，异戊烯化和随后的γ亚基甲基化/去甲基化是体外Gβγ-MT相互作用所必需的，所以在本研究中测试了靶向异戊烯化甲基化蛋白质甲基酯酶（PMPMEase）的小分子抑制剂（L-28和L-23）。我们发现这些抑制剂破坏了Gβγ和MT组织，并影响细胞形态和神经突生长。为了进一步支持Gβγ-MT相互作用在神经元分化中的作用，观察到在没有添加NGF的情况下，PC 12细胞中Gβγ的过表达诱导神经突生长。此外，过表达的Gβγ表现出与MT的结合模式，与在NGF分化细胞中观察到的相似。总之，我们的研究结果表明，异源三聚体G蛋白的βγ亚基通过与MT相互作用和调节MT重排在神经突起的生长和分化中起着关键作用。本文的在线版本（doi：10.1186/s12868-014-0132-4）包含补充材料，可供授权用户使用。
Assembly and disassembly of microtubules (MTs) is critical for neurite outgrowth and differentiation. Evidence suggests that nerve growth factor (NGF) induces neurite outgrowth from PC12 cells by activating the receptor tyrosine kinase, TrkA. G protein-coupled receptors (GPCRs) as well as heterotrimeric G proteins are also involved in regulating neurite outgrowth. However, the possible connection between these pathways and how they might ultimately converge to regulate the assembly and organization of MTs during neurite outgrowth is not well understood. Here, we report that Gβγ, an important component of the GPCR pathway, is critical for NGF-induced neuronal differentiation of PC12 cells. We have found that NGF promoted the interaction of Gβγ with MTs and stimulated MT assembly. While Gβγ-sequestering peptide GRK2i inhibited neurite formation, disrupted MTs, and induced neurite damage, the Gβγ activator mSIRK stimulated neurite outgrowth, which indicates the involvement of Gβγ in this process. Because we have shown earlier that prenylation and subsequent methylation/demethylation of γ subunits are required for the Gβγ-MTs interaction in vitro, small-molecule inhibitors (L-28 and L-23) targeting prenylated methylated protein methyl esterase (PMPMEase) were tested in the current study. We found that these inhibitors disrupted Gβγ and ΜΤ organization and affected cellular morphology and neurite outgrowth. In further support of a role of Gβγ-MT interaction in neuronal differentiation, it was observed that overexpression of Gβγ in PC12 cells induced neurite outgrowth in the absence of added NGF. Moreover, overexpressed Gβγ exhibited a pattern of association with MTs similar to that observed in NGF-differentiated cells. Altogether, our results demonstrate that βγ subunit of heterotrimeric G proteins play a critical role in neurite outgrowth and differentiation by interacting with MTs and modulating MT rearrangement. The online version of this article (doi:10.1186/s12868-014-0132-4) contains supplementary material, which is available to authorized users.
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