Staurosporine induces neurite outgrowth in neuronal hybrids (PC12EN) lacking NGF receptors

• Published in: Journal of cellular biochemistry Vol. 62; no. 3; pp. 356 - 371
• Main Authors: Rasouly, David, Shavit, Davidit, Zuniga, Ramiro, Elejalde, Rafael B., Unsworth, Brian R., Yayon, Avner, Lazarovici, Philip, Lelkes, Peter I.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.09.1996
• Subjects: Adrenal Medulla - cytology; Animals; basic fibroblast growth factor (bFGF); Biomarkers; Calcium-Calmodulin-Dependent Protein Kinases - drug effects; Calcium-Calmodulin-Dependent Protein Kinases - metabolism; Cattle; Cell Adhesion Molecules - drug effects; Cell Adhesion Molecules - metabolism; Cell Adhesion Molecules - pharmacology; Cell Differentiation - drug effects; endothelial cells; Enzyme Activation - drug effects; Enzyme Inhibitors - pharmacology; epidermal growth factor (EGF); Fibroblast Growth Factors - pharmacology; Focal Adhesion Kinase 1; Focal Adhesion Protein-Tyrosine Kinases; growth factor receptors; Growth Substances - pharmacology; Hybrid Cells - drug effects; Hybrid Cells - physiology; hybrids; Karyotyping; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Mitogens - pharmacology; nerve growth factor (NGF); Nerve Growth Factors; Nerve Tissue Proteins - drug effects; Nerve Tissue Proteins - metabolism; Neurites - drug effects; Neurites - physiology; neurotrophins; PC12 cells; PC12 Cells - drug effects; PC12 Cells - physiology; Phosphorylation; Protein-Tyrosine Kinases - antagonists & inhibitors; Protein-Tyrosine Kinases - drug effects; Protein-Tyrosine Kinases - metabolism; Rats; Receptor, Epidermal Growth Factor - biosynthesis; Receptor, Epidermal Growth Factor - drug effects; Receptors, Fibroblast Growth Factor - biosynthesis; Receptors, Fibroblast Growth Factor - drug effects; Receptors, Nerve Growth Factor - drug effects; Receptors, Nerve Growth Factor - metabolism; Signal Transduction; staurosporine; Staurosporine - pharmacology; Tyrosine - metabolism
• ISSN: 0730-2312; 1097-4644
• 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 p125???. 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. © 1996 Wiley‐Liss, Inc.