title: Treatment of intact striatal neurones with cholera toxin or 8-bromoadenosine 3',5'-(cyclic)phosphate decreases the ability of pertussis toxin to ADP-ribosylate the alpha-subunits of inhibitory and other guanine-nucleotide-binding regulatory proteins, Gi and Go. Evidence for two distinct mechanisms.
abstract: Using primary cultures of striatal neurones from the mouse embryo, we showed that treatment of intact cells with cholera toxin (5 micrograms/ml, 22 h) decreases the subsequent ADP-ribosylation of the alpha subunit of the guanine-nucleotide-binding regulatory protein Go (Go alpha) and the alpha subunit of the inhibitory guanine-nucleotide-binding regulatory protein (Gi alpha) of adenylate cyclase, which is catalyzed in vitro on neuronal membranes by pertussis toxin. The inhibitory effect of cholera toxin could not only be attributed to an increased production of cAMP in neurones. Treatment of cells with 0.1 microM 8-bromoadenosine 3',5'-(cyclic)phosphate (BrcAMP) for 16 h, or with 0.1 mM BrcAMP for 5 min, mimicked the effect of cholera toxin on the ADP-ribosylation of Go alpha and Gi alpha in vitro. However, the two agents seem to act through distinct mechanisms. The protein kinase inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine prevented the action of Br8cAMP but not that of cholera toxin. In addition, measurements of the pI of the Go alpha deduced from immunoblots of two-dimensional gels performed using a specific antibody directed against Go alpha suggest that treatment of neurones with cholera toxin induces ADP-ribosylation of Go alpha in intact cells, while BrcAMP does not.
authors.affiliation: Laboratoire de Neuropharmacologie, Institut National de la Santé et de la Recherche Médicale, Unité 114, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Maus
authors.forename: V
authors.initials: V
authors.lastname: Homburger
authors.forename: J
authors.initials: J
authors.lastname: Cordier
authors.forename: C
authors.initials: C
authors.lastname: Pantaloni
authors.forename: J
authors.initials: J
authors.lastname: Bockaert
authors.forename: J
authors.initials: J
authors.lastname: Glowinski
authors.forename: J
authors.initials: J
authors.lastname: Prémont
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D000244:Adenosine Diphosphate / Q000378:metabolism*; D037361:Adenylate Cyclase Toxin; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology*; D003342:Corpus Striatum / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D019204:GTP-Binding Proteins / Q000378:metabolism*; D015151:Immunoblotting; D007526:Isoelectric Point; D007546:Isoquinolines / Q000494:pharmacology; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D037342:Pertussis Toxin; D010879:Piperazines / Q000494:pharmacology; D012266:Ribose / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
chemical_list: D037361:Adenylate Cyclase Toxin; D007546:Isoquinolines; D010879:Piperazines; D010566:Virulence Factors, Bordetella; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000244:Adenosine Diphosphate; D012266:Ribose; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002772:Cholera Toxin; D000242:Cyclic AMP; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins