Abstract
Acetamiprid (ACE) and Imidacloprid (IMI) are widely-used neonicotinoid insecticides (NNIs) with functional activity at human acetylcholine nicotinic receptors and, therefore, with putative toxic effects. The objective of this study was the evaluation of the interactions between NNIs and α7-nAChR, as this receptor keeps intracellular Ca2+ ([Ca2+]i) to an optimum for an adequate neuronal functioning. Possible interactions between NNIs and the cryo-EM structure of the human α-7 nAChR were identified by molecular docking. Additionally, NNI effects were analyzed in neuroblastoma SH-SY5Y cells, as they naturally express α-7 nAChRs. Functional studies included proliferative/cytotoxic effects (MTT test) in undifferentiated SH-SY-5Y cells and indirect measurements of [Ca2+]i transients in retinoic acid-differentiated SH-SY-5Y cells loaded with Fluo-4 AM. Docking analysis showed that the binding of IMI and ACE occurred at the same aromatic cage that the specific α-7 nAChR agonist EVP-6124. IMI showed a better docking strength than ACE. According to the MTT assays, low doses (10–50 µM) of IMI better than ACE stimulated neuroblastoma cell proliferation. At higher doses (250–500 µM), IMI also prevailed over ACE and dose-dependently triggered more abrupt fluorescence changes due to [Ca2+]i mobilization in differentiated SH-SY5Y neurons. Indeed, only IMI blunted nicotine-evoked intracellular fluorescence stimulation (i.e., nicotine cross-desensitization). Summarizing, IMI demonstrated a superior docking strength and more robust cellular responses compared to ACE, which were likely associated with a stronger activity at α-7nAChRs. Through the interaction with α-7nAChRs, IMI would demonstrate its high neurotoxic potential for humans. More research is needed for investigating the proliferative effects of IMI in neuroblastoma cells.
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We are grateful to the Programa de las Naciones Unidas para el Desarrollo (PENUD) of ECUADOR for the sponsorship of the SENESCYT grant (PIC-18-INE-YACHAY-002).
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This work was supported by the Secretaria Nacional, Ciencia, Tecnología e Innovación (SENESCYT) of the Ecuadorian Government (Ref. PIC-18-INE-YACHAY-002) to Santiago Ballaz and Lenin Ramirez-Cando and by the Fondo Nacional de Desarrollo Científico y Tecnológico-Agencia Nacional de Investigación y Desarrollo of the Chilean Government (Ref. FONDECYT-ANID 1221080) to Luis Aguayo.
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L.J.R-C. and S.J.B. conceived and designed the experiments, analyzed and interpreted the data, contributed reagents, materials, analysis tools or data, wrote the paper. M.S.G-V. performed the experiments, wrote the paper. L.G.A. analyzed and interpreted the data, contributed reagents, materials, analysis tools and data.
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Guzman-Vallejos, M.S., Ramirez-Cando, L.J., Aguayo, L. et al. Molecular Docking Analysis at the Human α7-nAChR and Proliferative and Evoked-Calcium Changes in SH-SY5Y Cells by Imidacloprid and Acetamiprid Insecticides. Neurotox Res 42, 16 (2024). https://doi.org/10.1007/s12640-024-00697-0
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DOI: https://doi.org/10.1007/s12640-024-00697-0