A potential cost of evolving epibatidine resistance in poison frogs

Abstract

BackgroundSome poison arrow frogs sequester the toxin epibatidine as a defense against predators. We previously identified a single amino acid substitution (S108C) at a highly conserved site in a neuronal nicotinic acetylcholine receptor (nAChR) {beta}2 subunit that prevents epibatidine from binding to this receptor. When placed in a homologous mammalian nAChR this substitution minimized epibatidine binding but also perturbed acetylcholine binding, a clear cost. However, in the nAChRs of poison arrow frogs, this substitution appeared to have no detrimental effect on acetylcholine binding and, thus, appeared cost-free. ResultsThe introduction of S108C into the 4{beta}2 nAChRs of non-dendrobatid frogs also does not affect ACh sensitivity, when these receptors are expressed in Xenopus laevis oocytes. However, 4{beta}2 nAChRs with C108 had a decreased magnitude of neurotransmitter-induced currents in all species tested (Epipedobates anthonyi, non-dendrobatid frogs, as well as human), compared with 4{beta}2 nAChRs with the conserved S108. Immunolabeling of frog or human 4{beta}2 nAChRs in the plasma membrane using radiolabeled antibody against the {beta}2 nAChR subunit shows that C108 significantly decreased the number of cell-surface 4{beta}2 nAChRs, compared with S108. ConclusionsWhile S108C protects these species against sequestered epibatidine, it incurs a potential physiological cost of disrupted 4{beta}2 nAChR function. These results may explain the high conservation of a serine at this site in vertebrates, as well as provide an example of a tradeoff between beneficial and deleterious effects of an evolutionary change. They also provide important clues for future work on assembly and trafficking of this important neurotransmitter receptor.