Cholera toxin-B subunit blocks excitatory effects of opioids on sensory neuron action potentials indicating that GM1 ganglioside may regulate Gs-linked opioid receptor functions.
In a previous study, we demonstrated that cholera toxin-A subunit, as well as the whole toxin, selectively blocks opioid-induced prolongation of the Ca2+ component of the action potential duration (APD) in dorsal root ganglion (DRG) neurons, indicating mediation of this excitatory effect by Gs-linked opioid receptors. The present study shows that pretreatment of DRG neurons with the B subunit of cholera toxin (1-10 ng/ml; greater than 15 min) can also block mu/delta and kappa opioid-induced APD prolongation, but not shortening. Since the B subunit binds selectively to GM1 ganglioside located on the cell surface, these results suggest that this ganglioside may regulate Gs-linked excitatory opioid receptor functions in DRG neurons. Possible contamination of purified B subunit preparations of cholera toxin with traces of the more potent A subunit was eliminated by heating the stock solution to 56 degrees C for 20 min. Exposure of DRG neurons to an affinity-purified anti-GM1 antiserum also blocked opioid-induced APD prolongation, providing further evidence that GM1 ganglioside may play an essential role in excitatory opioid modulation of the action potential of these cells. The blockade by cholera toxin-B subunit and anti-GM1 antibodies of opioid-induced APD prolongation is best accounted for by the following hypothesis: CTX-B interferes with an endogenous GM1 ganglioside component of the excitatory, but not inhibitory, opioid receptor complex on DRG neurons that may allosterically regulate coupling of the receptors via Gs to adenylate cyclase/cyclic adenosine monophosphate-dependent ionic conductances.
Shen
K F
KF
Department of Neuroscience, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY 10461.
Crain
S M
SM
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002772:Cholera Toxin / Q000494:pharmacology*; D004399:Dynorphins / Q000494:pharmacology*; D005333:Fetus; D005677:G(M1) Ganglioside / Q000276:immunology / Q000502:physiology*; D005727:Ganglia, Spinal / Q000187:drug effects / Q000502:physiology*; D007106:Immune Sera; D051379:Mice; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology*; D009924:Organ Culture Techniques; D011957:Receptors, Opioid / Q000502:physiology*
D007106:Immune Sera; D011957:Receptors, Opioid; D005677:G(M1) Ganglioside; D004399:Dynorphins; D002772:Cholera Toxin