Imidacloprid effects on acetylcholinesterase and nicotinic acetylcholine receptor in Apis mellifera. Experimental and molecular modeling approaches

• Published in: Chemosphere (Oxford) Vol. 356; p. 141899
• Main Authors: Ali, Hussein M., Abdel-Aty, Basma, El-Sayed, Walaa, Mariy, Faiza M., Hegazy, Gamal M., Mohamed, Rehab A., Zoghly, Hala M.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-05-2024
• Subjects: AChE; Alfa subunit; Honeybee; nAChR; Neonicotinoids; Transmembranes; AChE; Honeybee; AC; nAChRα1; IMI; CCD; ACh; Neonicotinoids; ES; nAChR; Alfa subunit; Transmembranes; TM
• ISSN: 0045-6535; 1879-1298
• DOI: 10.1016/j.chemosphere.2024.141899

Although the neonicotinoid insecticides have good selectivity towards insects rather than vertebrates, they have severe effects on honeybee production and pollination activities. Therefore, the effects of imidacloprid (IMI), the most used neonicotinoid, on the two main bioreceptors, acetylcholinesterase (AChE) and nicotinic acetylcholine receptor alpha subunit (nAChRα1) of honeybees were examined to identify their roles in honeybee toxicity and possible binding sites which assist in selecting and designing neonicotinoids. In vivo, IMI showed a high inhibitory effect on AChE (IC50 5.63 mg/L); however, the effect was much lower in vitro experiment (IC50 719 mg/L). This result induced us to examine the IMI effect on AChE gene expression which revealed that the AChE-2 gene expression was severely affected by IMI explaining the observed high enzyme inhibition. In addition, although toxicity increased by increasing exposure to IMI (LC50 2.9 mg/L after 4h and 0.75 mg/L after 48h), AChE was not elevated (IC50 5.63 and 5.52 mg/L respectively). Besides, Despite resuming most enzyme activity (77% during 2 h and 84.14% after 4 h), a high mortality level was observed with LC50 2.9 mg/L. These results reinforced that the observed high toxicity is a multifactor process. Accordingly, Molecular modeling and docking of IMI into honeybee AChE and nAChRα1were also performed to examine their possible interactions and identify the important binding sites. Results models indicated that the first two binding sites in AChE were found in the esteratic subunit in the active site explaining the observed in vitro inhibition. In nAChRα1, four of the highest five free energy binding sites are located in the large TM3-TM4 loop and one in the extracellular loops. Consequently, the present work revealed that IMI toxicity is attributed to various factors including direct interaction with both AChE and nAChRα1 as well as downregulating AChE-2 gene expression.
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•AChE was inhibited by IMI both in vivo (IC50 5.63 ppm) and in vitro (IC50 710.06 ppm).•Increasing IMI exposure time increased toxicity but did not increase AChE inhibition.•AChE undergoes fast recovery in vivo (84.14% in 4h).•AChE inhibition is due to both direct interaction and suppression of gene expression.•The role of each IMI structural moiety in nAChRα1 binding and binding stes were identified.