Rasouly D, Shavit D, Zuniga R, Elejalde R B, Unsworth B R, Yayon A, Lazarovici P, Lelkes P I
Department of Pharmacology and Experimental Therapeutics, School of Pharmacy, Faculty of Medicine, Hebrew University of Jerusalem, Israel.
J Cell Biochem. 1996 Sep 1;62(3):356-71. doi: 10.1002/(sici)1097-4644(199609)62:3<356::aid-jcb6>3.0.co;2-q.
A novel neuronal model (PC12EN cells), obtained by somatic hybridization of rat adrenal medullary pheochromocytoma (PC12) and bovine adrenal medullary endothelial (BAME) cells, was developed. PC12EN cells maintained numerous neuronal characteristics: they expressed neuronal glycolipid conjugates, synthesized and secreted catecholamines, and responded to differentiative agents with neurite outgrowth. PC12EN lacked receptors for EGF and both the p75 and trk NGF receptors, while FGF receptor expression was maintained. Staurosporine (5-50 nM), but not other members of the K252a family of protein kinase inhibitors, rapidly induced neurite outgrowth in PC12EN, as also found in the parental PC12 cells, but not in BAME cells. Similarly, both acidic and basic FGF (1-100 ng/ml) were neurotropic in PC12EN. In contrast to the mechanism by which FGF promoted neurite outgrowth in PC12EN, the neurotropic effect of staurosporine did not involve activation of established signalling pathways, such as tyrosine phosphorylation of erk (ras pathway) or SNT (a specific target of neuronal differentiation). In addition, staurosporine induced the tyrosine phosphorylation of the focal adhesion kinase p125FAK. However, since the latter effect was also observed with other protein kinase inhibitors of the K252a family, which induced PC12EN cells flattening but no neurite extension, we propose that FAK tyrosine phosphorylation may be related to ubiquitous changes in cell shape. We anticipate that PC12EN neuronal hybrids will become useful models in neuroscience research for evaluating unique cellular signalling mechanisms of novel neurotropic compounds.
通过大鼠肾上腺髓质嗜铬细胞瘤（PC12）与牛肾上腺髓质内皮（BAME）细胞进行体细胞杂交，获得了一种新型神经元模型（PC12EN细胞）。PC12EN细胞保持了许多神经元特征：它们表达神经元糖脂缀合物，合成并分泌儿茶酚胺，并且对分化剂产生神经突生长反应。PC12EN缺乏表皮生长因子（EGF）受体以及p75和trk神经生长因子（NGF）受体，而成纤维细胞生长因子（FGF）受体表达得以保留。星形孢菌素（5 - 50 nM），而非K252a家族蛋白激酶抑制剂的其他成员，能快速诱导PC12EN细胞长出神经突，这在亲代PC12细胞中也有发现，但在BAME细胞中未出现。同样，酸性和碱性FGF（1 - 100 ng/ml）对PC12EN细胞具有神经营养作用。与FGF促进PC12EN细胞神经突生长的机制不同，星形孢菌素的神经营养作用并不涉及激活既定信号通路，如细胞外信号调节激酶（erk，ras通路）的酪氨酸磷酸化或SNT（神经分化的特定靶点）。此外，星形孢菌素诱导粘着斑激酶p125FAK的酪氨酸磷酸化。然而，由于在K252a家族的其他蛋白激酶抑制剂处理时也观察到了后者的效应，这些抑制剂会使PC12EN细胞变平但不会诱导神经突延伸，因此我们认为FAK酪氨酸磷酸化可能与细胞形状的普遍变化有关。我们预计PC12EN神经元杂交细胞将成为神经科学研究中评估新型神经营养化合物独特细胞信号机制的有用模型。
