Involvement of the neurotensin receptor-3 in the neurotensin-induced migration of human microglia
Involvement of the neurotensin receptor-3 in the neurotensin-induced migration of human microglia
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DOI：
10.1523/jneurosci.23-04-01198.2003
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发表时间：
2003-02-15
影响因子：
5.3
通讯作者：
Mazella, J
中科院分区：
文献类型：
--
作者：
Martin, S;Vincent, JP;Mazella, J
小胶质细胞的运动性在应对脑组织损伤或胞毒损伤中起着至关重要的作用。我们使用了两种体外实验，创伤愈合模型和趋化实验，证明神经肽神经降压素通过依赖于磷脂酰肌醇3-激酶(PI3-激酶)和丝裂原激活蛋白(MAP)激酶途径来诱导人小胶质细胞系C13NJ迁移。神经降压素对细胞迁移的影响被神经降压素受体-3前肽所阻断，神经降压素受体-3是神经降压素受体的选择性配体。通过RT-PCR、光亲和标记和Western印迹分析，我们证明了I型神经降压素受体-3是这些小胶质细胞中唯一已知的神经降压素受体，它的激活导致了细胞外信号调节激酶1/2和Akt的磷酸化。此外，在神经降压素对细胞迁移的影响之前，F-肌动蛋白细胞骨架发生了深刻的改变，特别是通过迅速形成大量的细胞丝状伪足。神经降压素诱导的运动和丝状足的出现均可被MAP激酶或PI3-激酶通路的选择性抑制剂完全阻断。这表明神经降压素受体-3具有功能，并介导神经降压素的迁移作用。
Microglia motility plays a crucial role in response to lesion or exocytotoxic damage of the cerebral tissue. We used two in vitro assays, a wound-healing model and a chemotaxis assay, to show that the neuropeptide neurotensin elicited the migration of the human microglial cell line C13NJ by a mechanism dependent on both phosphatidylinositol 3-kinase (PI3-kinase) and mitogen-activated protein (MAP) kinase pathways. The effect of neurotensin on cell migration was blocked by the neurotensin receptor-3 propeptide, a selective ligand of this receptor. We demonstrate, by using RT-PCR, photoaffinity labeling, and Western blot analysis, that the type I neurotensin receptor-3 was the only known neurotensin receptor expressed in these microglial cells and that its activation led to the phosphorylation of both extracellular signal-regulating kinases 1/2 and Akt. Furthermore, the effect of neurotensin on cell migration was preceded by a profound modification of the F-actin cytoskeleton, particularly by the rapid formation of numerous cell filopodia. Both the motility and the filopodia appearance induced by neurotensin were totally blocked by selective inhibitors of MAP kinases or PI3-kinase pathways. This demonstrates that the neurotensin receptor-3 is functional and mediates the migratory actions of neurotensin.