Axonal growth regulation of fetal and embryonic stem cell-derived dopaminergic neurons by Netrin-1 and Slits
Axonal growth regulation of fetal and embryonic stem cell-derived dopaminergic neurons by Netrin-1 and Slits
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DOI：
10.1634/stemcells.2006-0119
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发表时间：
2006-11-01
期刊：
影响因子：
5.2
通讯作者：
Isacson, Ole
中科院分区：
文献类型：
--
作者：
Lin, Ling;Isacson, Ole
通过植入胚胎干细胞对帕金森病中受损或丢失的多巴胺回路进行物理修复可能成为一种治疗方法。了解ES衍生的多巴胺(DA)神经元对决定轴突路径和靶向的引导信号的反应是至关重要的。应用胶原凝胶培养系统，观察分泌分子Netrin-1和Slits对胚胎DA神经元和ES分化的DA神经元突起生长的影响。我们以前已经证明，胎儿DA神经元表达DCC和Robo1/2受体，Netrin-1和Slit2对DA突起的生长具有吸引和排斥作用。在本研究中，我们观察到Slit1和SLIT3都排斥和抑制胎儿DA神经元的轴突生长。在这里，我们还证明了ES分化的神经元，包括DA神经元，表达Netrin受体DCC和Sit受体Robo蛋白。在ES细胞凝胶培养体系中，Netrin-1通过DCC受体促进突起生长，Slit1和SLIT3通过Robo受体抑制突起生长。Slit2在共培养实验中表现出抑制和排斥作用。然而，与胎儿DA突起不同的是，在ES来源的DA神经元与Netrin-1、Slit1和SLIT3产生细胞的共同培养中，没有观察到定向突起生长。这些发现表明，目前的方法产生的胚胎来源的DA神经元在体外可以以类似于胎儿细胞的方式对指导信号做出反应，但也表现出不同的反应。这可能是由于胚胎干细胞分化过程中现有的体外细胞构型或条件处理方法产生的发育差异所致。
The physical restoration of dopamine circuits damaged or lost in Parkinson disease by implanting embryonic stem (ES)-derived cells may become a treatment. It is critical to understand responses of ES-derived dopamine (DA) neurons to guidance signals that determine axonal path and targeting. Using a collagen gel culture system, we examined effects of secreted molecules Netrin-1 and Slits on neurite outgrowth of fetal DA neurons and murine ES-differentiated DA neurons. We have previously shown that fetal DA neurons express DCC and Robo1/2 receptors and that Netrin-1 and Slit2 function as an attractant and a repellent for DA neurite outgrowth. In the present study, we observe that both Slit1 and Slit3 repel and inhibit neurite growth of fetal DA neurons. Here, we also demonstrate that ES-differentiated neurons including DA neurons express the Netrin receptor DCC and Slit receptor Robo proteins. In the gel culture system of ES cells, Netrin-1 promoted neurite outgrowth mediated by DCC receptor, and Slit1 and Slit3 were inhibitory for neurite outgrowth through Robo receptors. Slit2 appeared to exert inhibitory as well as repulsive effects in the coculture assay. However, unlike fetal DA neurites, no directed neurite outgrowth was observed in the cocultures of ES-derived DA neurons with Netrin-1-, Slit1-, and Slit3-producing cells. The findings suggest that ES-derived DA neurons generated by current protocols can respond to guidance cues in vitro in a similar manner to fetal cells but also exhibit distinct responses. This may result from developmental differences generated by present in vitro methods of cell patterning or conditioning during ES cell differentiation.