COUP-TFI (Chicken ovalbumin upstream promoter-transcription factor I) regulates cell migration and axogenesis in differentiating P19 embryonal carcinoma cells
COUP-TFI (Chicken ovalbumin upstream promoter-transcription factor I) regulates cell migration and axogenesis in differentiating P19 embryonal carcinoma cells
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DOI：
10.1210/me.14.12.1918
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发表时间：
2000-12-01
影响因子：
--
通讯作者：
Salbert, G
中科院分区：
文献类型：
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作者：
Adam, F;Sourisseau, T;Salbert, G
核孤儿受体COUP-TFs（鸡卵白蛋白上游启动子转录因子）的发育表达模式与几种动物的神经发生有关。然而，coup - tf在神经发生中的作用仍然未知。我们通过两种互补的方法研究了COUP-TFI在维甲酸（RA）诱导的猪胚胎癌细胞神经元分化中的功能参与：1)去调控COUP-TFI的表达，以及2)通过显性阴性的COUP-TFI突变体使内源性COUP-TFI失活。低水平的野生型（wt）COUP-TFI转基因表达不抑制神经细胞的命运，并主要增强了ra处理的P19聚集体的神经元生长。相反，高COUP-TFI表达阻碍了RA诱导的猪细胞的神经元分化，导致细胞培养缺乏神经元。这种形态学效应与e -钙粘蛋白mRNA水平升高有关。显性阴性的COUP-TFI突变体在类风湿关节炎治疗后诱导细胞堆积，抑制神经突的延伸和神经元的生长。RGD肽干扰实验表明内源性coup - tf通过整合素依赖机制促进神经元迁移。因此，RT-PCR分析显示，COUP-TFI上调了玻璃体连接蛋白mRNA水平，并且在瞬时转染实验中，COUP-TFI刺激了小鼠玻璃体连接蛋白启动子的活性。综上所述，这些数据表明，COUP-TFI不仅仅是类视黄醛功能的全局抑制因子，而且在调节参与细胞粘附和迁移过程的基因方面表现出高选择性，这对神经元分化尤为重要。
The developmental expression patterns of the nuclear orphan receptors COUP-TFs (chicken ovalbumin upstream promoter-transcription factors) have been correlated to neurogenesis in several animal species. Nevertheless, the role of COUP-TFs in neurogenesis remains unknown. We have studied the functional involvement of COUP-TFI in retinoic acid (RA)-induced neuronal differentiation of pig embryonal carcinoma cells through two complementary approaches: 1) deregulated expression of COUP-TFI, and 2) inactivation of endogenous COUP-TFs by means of a dominant-negative COUP-TFI mutant. Low levels of wild-type (wt)COUP-TFI transgene expression did not inhibit neural cell fate and primarily enhanced neuron outgrowth from RA-treated P19 aggregates. In contrast, high COUP-TFI expression impeded the neuronal differentiation of pig cells induced with RA, resulting in cell cultures lacking neurons. This morphological effect was correlated to an elevated level of E-cadherin mRNA. The dominant-negative COUP-TFI mutant induced cell packing after RA treatment and inhibited neurite extension and neuron outgrowth from aggregates. A RGD peptide interference assay indicated that endogenous COUP-TFs could favor migration of neurons through an integrin-dependent mechanism. Accordingly, vitronectin mRNA levels were shown to be up-regulated by COUP-TFI by RT-PCR analysis, and COUP-TFI stimulated the mouse vitronectin promoter activity in transient transfection assays. Taken together, these data indicate that COUP-TFI is not simply a global repressor of retinoid functions, but shows a high selectivity for regulating genes involved in cellular adhesion and migration processes that are particularly important for neuronal differentiation.