Neurite outgrowth is enhanced by anti-idiotypic monoclonal antibodies to the ganglioside GM1.
Exogenously added gangliosides enhance sprouting, neurite outgrowth, and other neuronal activities; this effect may be initiated when a ganglioside binds to a membrane protein or when a ganglioside intercalates into the plasma membrane. To test whether binding to membrane proteins is sufficient for ganglioside-mediated activity, anti-idiotypic antibodies were generated that mimic the functional binding sites of the ganglioside GM1 as described by M. J. Riggott and W. D. Matthew (1996, Glycobiology, 6, 581-589). These anti-idiotypic antibodies are proteinaceous probes that model the biochemical and biological effects of gangliosides. Those anti-idiotypic ganglioside (AIG) monoclonal antibodies (mAb's) were selected based on their ability to bind a known GM1 binding protein, the beta-subunit of cholera toxin. These studies described neuronal cell surface proteins that were identified by immunocytochemistry and Western blotting using these AIG mAb's. Here we show that AIG mAb's mimic the functional properties of GM1 in that they facilitate neurite outgrowth from central and peripheral nervous system neurons in in vitro bioassays. In addition, AIG mAb binding modulates second messenger activity, suggesting that membrane protein binding alone is sufficient to invoke intracellular activation. The similarity in the pattern of protein tyrosine phosphorylation evoked by GM1 and the anti-idiotypic ganglioside antibodies suggests that the AIG mAb's modulate neurite outgrowth in a manner similar to that of GM1. Because antibodies cannot intercalate into the plasma membrane, these results suggest that the ganglioside GM1 can mediate neuronal cellular activity by binding to cell surface proteins.
Riggott
M J
MJ
Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710, USA. riggottm@ohsu.edu
Matthew
W D
WD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000937:Antigen-Antibody Reactions / Q000502:physiology; D002478:Cells, Cultured; D003714:Denervation; D005260:Female; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005677:G(M1) Ganglioside / Q000032:analysis / Q000276:immunology*; D005727:Ganglia, Spinal / Q000166:cytology; D006624:Hippocampus / Q000166:cytology; D009416:Nerve Regeneration / Q000502:physiology; D016501:Neurites / Q000737:chemistry / Q000276:immunology*; D010766:Phosphorylation; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000601:surgery; D015290:Second Messenger Systems / Q000502:physiology; D014443:Tyrosine / Q000502:physiology
D000911:Antibodies, Monoclonal; D005452:Fluoresceins; D005456:Fluorescent Dyes; C024098:6-carboxyfluorescein; D005677:G(M1) Ganglioside; D014443:Tyrosine