Human muscarinic acetylcholine receptors are a target of the marine toxin 13-desmethyl C spirolide.
Spirolides are a group of cyclic imine marine toxins recently described. Although no human intoxication has been related to their presence in shellfish yet, the possible toxicological consequences to human health are actually unknown. The elucidation of the spirolide mechanism/s of action would help to estimate the threat to human consumers. Previous toxicological studies in mice suggested the involvement of acetylcholine receptors. In this work, the effects of the 13-desmethyl C spirolide on the activity and the expression of muscarinic acetylcholine receptors (mAChR) were analyzed using a human neuroblastoma cell model. The 13-desmethyl C spirolide inhibited the acetylcholine-induced calcium signal with a reduction of the maximum response to acetylcholine in the presence of the toxin. The 13-desmethyl C spirolide also reduced binding of the mAChR specific antagonist [(3)H]QNB to neuroblastoma cells. The effect of the 13-desmethyl C spirolide persisted after toxin removal and was inhibited by protection of the primary binding site with high concentrations of atropine suggesting an interaction of the spirolide with the orthologous binding site of mAChR. Moreover, the toxin induced a change in the characteristics of the membrane-associated M3 mAChRs, although it did not alter the total levels of M3 mAChR protein. The 13-desmethyl C spirolide targets mAChRs causing a reduction of function, a decrease of specific antagonist binding to mAChRs, and alteration of membrane-bound receptors that might have important toxicological implications.
Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, Campus Universitario, Lugo, Spain.
Carolina B
CB
Wandscheer
Natalia
N
Vilariño
Begoña
B
Espiña
M Carmen
MC
Louzao
Luis M
LM
Botana
D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D045744:Cell Line, Tumor; D006801:Humans; D008387:Marine Toxins / Q000737:chemistry / Q000633:toxicity; D011485:Protein Binding; D011976:Receptors, Muscarinic / Q000378:metabolism*; D013141:Spiro Compounds / Q000737:chemistry / Q000633:toxicity*
C517170:13-desmethylspirolide C; D008387:Marine Toxins; D011976:Receptors, Muscarinic; D013141:Spiro Compounds; D002118:Calcium