Distinct targets for tetanus and botulinum A neurotoxins within the signal transducing pathway in chromaffin cells.
Tetanus and botulinum A neurotoxins inhibited exocytosis evoked by various secretagogues in intact and permeabilized chromaffin cells. The block of exocytosis in intact chromaffin cells due to botulinum A neurotoxin could partially be overcome by enhancing nicotine- and veratridine-induced stimulation, whereas the block due to tetanus toxin persisted under the same conditions. The receptor-mediated restoration of 3H-noradrenaline release was specific for nicotinic stimulation, because exocytosis did not occur during muscarinic stimulation. Depolarization of intact chromaffin cells with increasing concentration of K+ failed to restore exocytosis that had been blocked by either toxin. When chromaffin cells, treated with tetanus or botulinum A neurotoxins, were exposed to the Ca2(+)-ionophore A 23187 or permeabilized by staphylococcal alpha-toxin, Ca2(+)-stimulated exocytosis was also inhibited. The inhibition was unaffected by increasing concentrations of free Ca2+. Activation of proteinkinase C and of G-proteins by phorbolester and GMPPNHP, respectively, increased Ca2(+)-induced exocytosis in control cells as well as in cells treated with tetanus and botulinum A neurotoxins. The block, however, could not be relieved by these manipulations, and it could not be relieved by activating the cGMP or cAMP pathways with analoga of cyclic nucleotides, phosphodiesterases inhibitors, and forskolin either. It is concluded that nicotine and veratridine trigger a mechanism within the sequence of events leading to exocytosis that is located beyond the increase in intracellular Ca2(+)-concentration. This pathway may not be affected by botulinum A neurotoxin. The target of tetanus toxin is probably located even closer to the fusion process, i.e. beyond the step upon which botulinum A neurotoxin acts.
Marxen
P
P
Institut für Peptidforschung, Hannover, Federal Republic of Germany.
Bartels
F
F
Ahnert-Hilger
G
G
Bigalke
H
H
D000311:Adrenal Glands / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology; D001905:Botulinum Toxins / Q000633:toxicity*; D002118:Calcium / Q000502:physiology; D002395:Catecholamines / Q000378:metabolism; D002417:Cattle; D002838:Chromaffin System / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D005089:Exocytosis / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism; D006460:Hemolysin Proteins; D006461:Hemolysis / Q000187:drug effects; D066298:In Vitro Techniques; D009498:Neurotoxins / Q000633:toxicity*; D009538:Nicotine / Q000494:pharmacology; D009638:Norepinephrine / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013744:Tetanus Toxin / Q000633:toxicity*
D001427:Bacterial Toxins; D002395:Catecholamines; D006460:Hemolysin Proteins; D009498:Neurotoxins; D013744:Tetanus Toxin; C021091:staphylococcal alpha-toxin; D009538:Nicotine; D000242:Cyclic AMP; D011493:Protein Kinase C; D001905:Botulinum Toxins; D019204:GTP-Binding Proteins; D006152:Cyclic GMP; D011188:Potassium; D002118:Calcium; D009638:Norepinephrine