Staurosporine induces neurite outgrowth in neuronal hybrids (PC12EN) lacking NGF receptors.
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.
Rasouly
D
D
Department of Pharmacology and Experimental Therapeutics, School of Pharmacy, Faculty of Medicine, Hebrew University of Jerusalem, Israel.
Shavit
D
D
Zuniga
R
R
Elejalde
R B
RB
Unsworth
B R
BR
Yayon
A
A
Lazarovici
P
P
Lelkes
P I
PI
D000313:Adrenal Medulla / Q000166:cytology; D000818:Animals; D015415:Biomarkers; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000187:drug effects / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000096:biosynthesis / Q000187:drug effects; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006133:Growth Substances / Q000494:pharmacology; D006822:Hybrid Cells / Q000187:drug effects / Q000502:physiology*; D007621:Karyotyping; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D008934:Mitogens / Q000494:pharmacology; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000502:physiology*; D016716:PC12 Cells / Q000187:drug effects / Q000502:physiology*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000096:biosynthesis / Q000187:drug effects; D017475:Receptors, Nerve Growth Factor / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction; D019311:Staurosporine / Q000494:pharmacology*; D014443:Tyrosine / Q000378:metabolism
D015415:Biomarkers; D015815:Cell Adhesion Molecules; D004791:Enzyme Inhibitors; D006133:Growth Substances; D008934:Mitogens; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D017468:Receptors, Fibroblast Growth Factor; D017475:Receptors, Nerve Growth Factor; D014443:Tyrosine; D005346:Fibroblast Growth Factors; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496052:Ptk2 protein, rat; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D019311:Staurosporine