Muscarinic receptor-mediated induction of actin-driven lamellar protrusions in neuroblastoma cell somata and growth cones. Involvement of protein kinase C.
In SH-SY5Y human neuroblastoma cells, addition of acetylcholine or carbachol rapidly induces a transient protrusion of lamellipodia. The protrusions appear after a delay of 30 sec and persist for a period of about 5 min at the margins of cell somata and at the distal parts of cell processes. They are caused by a strikingly increased, cytochalasin B-sensitive assembly of actin at the cell periphery. They often detach from the substrate, retracting and protruding again. In retinoic acid-induced neuronally differentiated cells, this initialized protrusive activity is restricted to growth cones. d-Tubocurarine does not influence, but atropine totally inhibits the cholinergic induction of the actin-driven protrusions, suggesting that a muscarinic receptor-mediated activation of the phosphoinositol signaling pathway is involved. Depolarization by increase of the potassium concentration and ionophore-mediated Ca(2+)-influx are ineffective to trigger the protrusive and ruffling activity. An identical cytochalasin B-sensitive actin-driven response is caused by treating of the cells with the protein kinase C (PKC) activator 12-myristate-13-acetate. In this case, however, lamellar protrusions are formed after a delay of at least 3 min and are maintained for several days. Incubating the cells with the protein kinase C inhibitor bisindolylmaleide or staurosporine inhibits both the muscarinic receptor-mediated and phorbolester-mediated actin-driven response, suggesting that activated PKC plays a crucial role.
Rösner
H
H
Institute of Zoology, University of Hohenheim, Stuttgart, Germany.
Vacun
G
G
Rebhan
M
M
D000109:Acetylcholine / Q000494:pharmacology; D000199:Actins / Q000037:antagonists & inhibitors / Q000378:metabolism; D000470:Alkaloids / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D048429:Cell Size; D003470:Culture Media; D003571:Cytochalasin B / Q000494:pharmacology; D005456:Fluorescent Dyes; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D016501:Neurites / Q000201:enzymology* / Q000648:ultrastructure; D009447:Neuroblastoma / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011976:Receptors, Muscarinic / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000648:ultrastructure
D000199:Actins; D000470:Alkaloids; D003470:Culture Media; D005456:Fluorescent Dyes; D007211:Indoles; D008301:Maleimides; D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic; D003571:Cytochalasin B; D002217:Carbachol; D011493:Protein Kinase C; D019311:Staurosporine; C088060:bisindolylmaleimide; D000109:Acetylcholine; D013755:Tetradecanoylphorbol Acetate