Chai Xuejun, Förster Eckart, Zhao Shanting, Bock Hans H, Frotscher Michael
Institut für Anatomie und Zellbiologie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg, Germany.
J Neurosci. 2009 Jan 7;29(1):288-99. doi: 10.1523/JNEUROSCI.2934-08.2009.
The extracellular matrix protein Reelin, secreted by Cajal-Retzius cells in the marginal zone of the cortex, controls the radial migration of cortical neurons. Reelin signaling involves the lipoprotein receptors apolipoprotein E receptor 2 (ApoER2) and very low density lipoprotein receptor (VLDLR), the adapter protein Disabled1 (Dab1), and phosphatidylinositol-3-kinase (PI3K). Eventually, Reelin signaling acts on the cytoskeleton; however, these effects on cytoskeletal organization have remained elusive. In Reelin-deficient mutant mice, most cortical neurons are unable to migrate to their destinations, suggesting a role for Reelin signaling in the dynamic cytoskeletal reorganization that is required for neurons to migrate. Here, we show that Reelin signaling leads to serine3 phosphorylation of n-cofilin, an actin-depolymerizing protein that promotes the disassembly of F-actin. Phosphorylation at serine3 renders n-cofilin unable to depolymerize F-actin, thereby stabilizing the cytoskeleton. We provide evidence for ApoER2, Dab1, Src family kinases (SFKs), and PI3K to be involved in n-cofilin serine3 phosphorylation. Phosphorylation of n-cofilin takes place in the leading processes of migrating neurons as they approach the Reelin-containing marginal zone. Immunostaining for phospho-cofilin in dissociated reeler neurons is significantly increased after incubation in Reelin-containing medium compared with control medium. In a stripe choice assay, neuronal processes are stable on Reelin-coated stripes but grow on control stripes by forming lamellipodia. These novel findings suggest that Reelin-induced stabilization of neuronal processes anchors them to the marginal zone which appears to be required for the directional migration process.
细胞外基质蛋白Reelin由皮质边缘区的Cajal-Retzius细胞分泌,可控制皮质神经元的径向迁移。Reelin信号传导涉及脂蛋白受体载脂蛋白E受体2(ApoER2)和极低密度脂蛋白受体(VLDLR)、衔接蛋白Disabled1(Dab1)和磷脂酰肌醇-3-激酶(PI3K)。最终,Reelin信号传导作用于细胞骨架;然而,这些对细胞骨架组织的影响仍不明确。在Reelin缺陷型突变小鼠中,大多数皮质神经元无法迁移到其目的地,这表明Reelin信号传导在神经元迁移所需的动态细胞骨架重组中发挥作用。在这里,我们表明Reelin信号传导导致n-丝切蛋白的丝氨酸3磷酸化,n-丝切蛋白是一种肌动蛋白解聚蛋白,可促进F-肌动蛋白的解体。丝氨酸3磷酸化使n-丝切蛋白无法解聚F-肌动蛋白,从而稳定细胞骨架。我们提供了ApoER2、Dab1、Src家族激酶(SFK)和PI3K参与n-丝切蛋白丝氨酸3磷酸化的证据。n-丝切蛋白的磷酸化发生在迁移神经元接近含Reelin的边缘区时的前端。与对照培养基相比,在含Reelin的培养基中孵育后,解离的reeler神经元中磷酸化丝切蛋白的免疫染色显著增加。在条纹选择试验中,神经元突起在涂有Reelin的条纹上稳定,但在对照条纹上通过形成片状伪足生长。这些新发现表明,Reelin诱导的神经元突起稳定将它们锚定在边缘区,这似乎是定向迁移过程所必需的。