Receptors of the Glial Cell Line-Derived Neurotrophic Factor Family of Neurotrophic Factors Signal Cell Survival through the Phosphatidylinositol 3-Kinase Pathway in Spinal Cord Motoneurons
Receptors of the Glial Cell Line-Derived Neurotrophic Factor Family of Neurotrophic Factors Signal Cell Survival through the Phosphatidylinositol 3-Kinase Pathway in Spinal Cord Motoneurons
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DOI：
10.1523/jneurosci.19-21-09160.1999
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发表时间：
1999-11
期刊：
影响因子：
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通讯作者：
R. Soler;X. Dolcet;M. Encinas;J. Egea;J. Bayascas;J. Comella
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文献类型：
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作者：
R. Soler;X. Dolcet;M. Encinas;J. Egea;J. Bayascas;J. Comella
神经营养因子（GDNF）家族的成员（GDNF、neurturin、persephin和artemin）能够促进包括脊髓运动神经元在内的不同神经元群的体内和体外存活。这些因子通过多组分受体发出信号，这些多组分受体包括Ret受体酪氨酸激酶和GDNF家族受体α (GRFα)家族的一个成员，即糖基磷脂酰肌醇连接的辅受体。受体的激活诱导Ret磷酸化，从而导致促进生存的作用。Ret磷酸化可激活几种细胞内通路，但每种通路的激活所引起的生物学效应尚不清楚。在目前的工作中，我们描述了GDNF家族成员在培养中促进鸡运动神经元存活的能力。我们利用原位杂交和逆转录- pcr技术在鸡运动神经元中发现了Ret和GFRα-1、GFRα-2和GFRα-4的存在。通过Western blot分析和激酶实验，我们证明了这些因子能够诱导磷脂酰肌醇3激酶（PI 3激酶）和细胞外调节激酶(ERK) -丝裂原活化蛋白（MAP）激酶途径的激活。为了表征这些途径在生存效应中的作用，我们使用了PI 3-激酶抑制剂LY 294002和MAP激酶和ERK激酶（MEK）抑制剂PD 98059。我们证明LY 294002，而不是PD 98059，可以阻止GDNF-、neurturin-和persephin诱导的运动神经元存活，这表明PI 3-激酶细胞内通路介导了神经营养效应。
The members of the glial cell line-derived neurotrophic factor (GDNF) family of neurotrophic factors (GDNF, neurturin, persephin, and artemin) are able to promote in vivo and in vitro survival of different neuronal populations, including spinal cord motoneurons. These factors signal via multicomponent receptors that consist of the Ret receptor tyrosine kinase plus a member of the GDNF family receptor α (GRFα) family of glycosylphosphatidylinositol-linked coreceptors. Activation of the receptor induces Ret phosphorylation that leads the survival-promoting effects. Ret phosphorylation causes the activation of several intracellular pathways, but the biological effects caused by the activation of each of these pathways are still unknown. In the present work, we describe the ability of the GDNF family members to promote chicken motoneuron survival in culture. We show the presence of Ret and GFRα-1, GFRα-2, and GFRα-4 in chicken motoneurons using in situ hybridization and reverse transcription-PCR techniques. By Western blot analysis and kinase assays, we demonstrate the ability of these factors to induce the phosphatidylinositol 3 kinase (PI 3-kinase) and the extracellular regulated kinase (ERK)–mitogen-activated protein (MAP) kinase pathways activation. To characterize the involvement of these pathways in the survival effect, we used the PI 3-kinase inhibitor LY 294002 and the MAP kinase and ERK kinase (MEK) inhibitor PD 98059. We demonstrate that LY 294002, but not PD 98059, prevents GDNF-, neurturin-, and persephin-induced motoneuron survival, suggesting that PI 3-kinase intracellular pathway is responsible in mediating the neurotrophic effect.