title: Differentiation of Neuro-2a neuroblastoma cells by an antibody to GM3 ganglioside.
abstract: A monoclonal antibody against GM3 ganglioside (GM3Ab) was found to trigger differentiation of Neuro-2a cells in culture. The differentiation of Neuro-2a cells by GM3Ab was accompanied by increased levels of intracellular serotonin and amino acid neurotransmitters viz. aspartate, glutamate, glutamine, glycine and taurine. Further study indicated that the increase in the serotonin level was not due to a higher rate of serotonin synthesis but rather to a higher rate of active transport of serotonin from the medium. Studies on the cell surface gangliosides revealed that unlike the proliferating cells, the GM3Ab-mediated differentiated cells contained higher gangliosides in addition to GM3 and GM2 gangliosides. Analysis of total cellular proteins indicated the appearance of a 25 kDa protein, pI 5.4, in the GM3Ab-treated cells--a small amount of this protein was observed in dibutyryl cAMP (Bt2cAMP)-treated cells, however, the protein was totally absent in the 5-bromo-2'-deoxyuridine (BrdU)-treated cells. Investigation of the mode of action of GM3Ab indicated that the cellular differentiation was due to increased cAMP accumulation resulting from an increase in the adenylate cyclase activity. Further studies with different agents affecting protein kinase C (PKC) activity and direct assay of PKC ruled out the possibility that GM3Ab mediated its effect via PKC. This GM3Ab-induced differentiation could be inhibited by protein kinase A (PKA) inhibitor, H8, but could not be inhibited by sphingosine, an inhibitor of PKC. Pertussis toxin could mimic the effect of GM3Ab, suggesting that GM3Ab caused the elevation in the adenylate cyclase activity by reducing the Gi-protein inhibition of the adenylate cyclase. The data suggests that GM3Ab, after interaction with cell surface GM3, elevated intracellular cAMP level by withdrawing the inhibitory effect of some undefined factor(s) present in culture medium which normally keeps adenylate cyclase activity low through activation of Gi-protein.
authors.affiliation: Child Study Center, Yale University School of Medicine, New Haven, CT 06510.
authors.forename: D
authors.initials: D
authors.lastname: Chatterjee
authors.forename: M
authors.initials: M
authors.lastname: Chakraborty
authors.forename: G M
authors.initials: GM
authors.lastname: Anderson
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine / Q000494:pharmacology; D003994:Bucladesine / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004261:DNA Replication; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005679:G(M3) Ganglioside / Q000276:immunology / Q000502:physiology*; D005732:Gangliosides / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D009447:Neuroblastoma; D012701:Serotonin / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002395:Catecholamines; D005679:G(M3) Ganglioside; D005732:Gangliosides; D009363:Neoplasm Proteins; D012701:Serotonin; D003994:Bucladesine; D008715:Methionine; D001973:Bromodeoxyuridine; D013936:Thymidine