Functional expression and analysis of <i>Apis mellifera</i> α1 nicotinic acetylcholine receptor

GENG Junjie; WANG Yuquan; ZHANG Kun; WU Shaoying

doi:10.15886/j.cnki.rdswxb.20230140

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Volume 15 Issue 5

Sep. 2024

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Article Navigation > Journal of Tropical Biology > 2024 > 15(5): 623-631

GENG Junjie, WANG Yuquan, ZHANG Kun, WU Shaoying. Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor[J]. Journal of Tropical Biology, 2024, 15(5): 623-631. doi: 10.15886/j.cnki.rdswxb.20230140

Citation:

GENG Junjie, WANG Yuquan, ZHANG Kun, WU Shaoying. Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor[J]. Journal of Tropical Biology, 2024, 15(5): 623-631. doi: 10.15886/j.cnki.rdswxb.20230140

Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

doi: 10.15886/j.cnki.rdswxb.20230140

GENG Junjie^1,2,
WANG Yuquan^1,2,
ZHANG Kun^{1,2
,},
WU Shaoying^{1,2
,}

1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya, Hainan 572025, China;
2. School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China

Received Date: 2023-12-13
Rev Recd Date: 2024-02-07

Abstract

Neonicotinoids that act on insect nicotinic acetylcholine receptors（nAChRs） are common tools for crop pest control, but they have serious negative effects on pollinators such as Apis mellifera. In this study, the nAChR α1 subunit gene was cloned from A. mellifera and systematically analyzed. The interaction mechanism between the nAChR α1 subtype and neonicotinoids（imidacloprid, clothianidin, and dinotefuran） was investigated by using the two-electrode voltage clamp technique. Sequence alignment and phylogenetic tree analysis showed that the nAChR α1 subunit of A. mellifera had typical domain characteristics in the nAChR subunit family genes, and was most distantly related to the α1 subunit of Hemiptera insects and most closely related to the α1 subunit of bees from Hymenoptera. The combination of A. mellifera nAChR α1 subunit and Rattus norvegicus nAChR β2 subunit was co-expressed in Xenopus laevis oocytes. Electrophysiological results showed that imidacloprid had the highest agonistic efficiency and lowest agonistic affinity, that clothianidin had the lowest agonistic efficiency and that dinotefuran had the highest agonistic affinity for Amα1/rβ2 nAChR, indicating that the α1 subunit of A. mellifera nAChRs is one of the targets of neonicotinoids and the sensitivity of Amα1/rβ2 nAChR to different neonicotinoids is different. All these results have important theoretical significance for the development of novel highly selective insecticides for target insects.
- Apis mellifera,
- nicotinic acetylcholine receptor,
- α1 subunit,
- functional expression

References

[1]	LIU S, LIU Y, HE F, et al. Enantioselective olfactory effects of the neonicotinoid dinotefuran on honey bees (Apis mellifera L.)[J]. Journal of Agricultural and Food Chemistry, 2019, 67(43):12105-12116.
[2]	KLEIN A M, VAISSIÈRE B E, CANE J H, et al. Importance of pollinators in changing landscapes for world crops[J]. Proceedings Biological Sciences, 2007, 274(1608):303-313.
[3]	GALLAI N, SALLES J M, SETTELE J, et al. Economic valuation of the vulnerability of world agriculture confronted with pollinator decline[J]. Ecological Economics,2009, 68(3):810-821.
[4]	VANENGELSDORP D, UNDERWOOD R, CARON D, et al. Estimate of managed colony losses in the winter of 2006-2007:A report commissioned by the apiary inspectors of America[J]. American Bee Journal, 2007, 147:599-603.
[5]	AURELL D, BRUCKNER S, WILSON M, et al. A national survey of managed honey bee colony losses in the USA:results from the bee informed partnership for 2020-21 and 2021-22[J]. Journal of Apicultural Research, 2024, 63(1):1-14.
[6]	WILLIAMS G R, TARPY D R, VANENGELSDORP D, et al. Colony collapse disorder in context[J]. BioEssays:News and Reviews in Molecular, Cellular and Developmental Biology, 2010, 32(10):845-846.
[7]	VANENGELSDORP D, EVANS J D, SAEGERMAN C, et al. Colony collapse disorder:a descriptive study[J]. PLoS One, 2009, 4(8):e6481.
[8]	CHEN Y P, PETTIS J S, CORONA M, et al. Israeli acute paralysis virus:epidemiology, pathogenesis and implications for honey bee health[J]. PLoS Pathogens, 2014,10(7):e1004261.
[9]	MARTIN S J, HIGHFIELD A C, BRETTELL L, et al.Global honey bee viral landscape altered by a parasitic mite[J]. Science, 2012, 336(6086):1304-1306.
[10]	MESNAGE R, ANTONIOU M N. Ignoring adjuvant toxicity falsifies the safety profile of commercial pesticides[J].Frontiers in Public Health, 2018, 5:361.
[11]	LI G, ZHAO H, LIU Z, et al. The wisdom of honeybee defenses against environmental stresses[J]. Frontiers in Microbiology, 2018, 9:722.
[12]	STRAUB L, WILLIAMS G R, VIDONDO B, et al. Neonicotinoids and ectoparasitic mites synergistically impact honeybees[J]. Scientific Reports, 2019, 9(1):8159.
[13]	FORFERT N, TROXLER A, RETSCHNIG G, et al. Neonicotinoid pesticides can reduce honeybee colony genetic diversity[J]. PLoS One, 2017, 12(10):e0186109.
[14]	张伟,齐素贞,薛晓锋,等.农药对蜜蜂的影响及毒理学机制研究进展[J].农药学学报, 2022, 24(5):1125-1138.
[15]	LIU J, LI Y, ZHANG Z, et al. Low concentration of quercetin reduces the lethal and sublethal effects of imidacloprid on Apis cerana (Hymenoptera:Apidae)[J]. Journal of Economic Entomology, 2021, 114(3):1053-1064.
[16]	TOSI S, NIEH J C. A common neonicotinoid pesticide,thiamethoxam, alters honey bee activity, motor functions,and movement to light[J]. Scientific Reports, 2017, 7(1):15132.
[17]	TSVETKOV N, SAMSON-ROBERT O, SOOD K, et al.Chronic exposure to neonicotinoids reduces honey bee health near corn crops[J]. Science, 2017, 356(6345):1395-1397.
[18]	IHARA M, MATSUDA K. Neonicotinoids:molecular mechanisms of action, insights into resistance and impact on pollinators[J]. Current Opinion in Insect Science,2018, 30:86-92.
[19]	CHANGEUX J P. The nicotinic acetylcholine receptor:the founding father of the pentameric ligand-gated ion channel superfamily[J]. The Journal of Biological Chemistry, 2012, 287(48):40207-40215.
[20]	IHARA M, HIKIDA M, MATSUSHITA H, et al. Loops D, E and G in the Drosophila Dα1 subunit contribute to high neonicotinoid sensitivity of Dα1-chicken β2 nicotinic acetylcholine receptor[J]. British Journal of Pharmacology, 2018, 175(11):1999-2012.
[21]	李旭扬,陈舒苗,于津鹏,等. α6/α3β4烟碱型乙酰胆碱受体β4亚基双点突变体的制备及其功能研究[J].热带生物学报, 2021, 12(4):403-411.
[22]	马倩芸,江涛,于日磊.烟碱乙酰胆碱受体结构的研究进展[J].中国海洋药物, 2018, 37(2):97-102.
[23]	ZHOU Y, HAN Q, FENG K, et al. New binding sites of nicotinic acetylcholine receptors from Myzus persicae[J].Entomologia Generalis, 2023, 43(2):461-470.
[24]	LIU Z, WILLIAMSON M S, LANSDELL S J, et al. A nicotinic acetylcholine receptor mutation (Y151S) causes reduced agonist potency to a range of neonicotinoid insecticides[J]. Journal of Neurochemistry, 2006, 99(4):1273-1281.
[25]	DEDERER H, WERR M, ILG T. Differential sensitivity of Ctenocephalides felis and Drosophila melanogaster nicotinic acetylcholine receptor α1 and α2 subunits in recombinant hybrid receptors to nicotinoids and neonicotinoid insecticides[J]. Insect Biochemistry and Molecular Biology, 2011, 41(1):51-61.
[26]	WANG Z M, LI S, SHI C C, et al. The actions of neonicotinoid insecticides on nicotinic acetylcholine subunits Ldα1 and Ldα8 of Leptinotarsa decemlineata and assembled receptors[J]. Insect Science, 2022, 29(5):1387-1400.
[27]	GAUTHIER M. State of the art on insect nicotinic acetylcholine receptor function in learning and memory[J].Advances in Experimental Medicine and Biology, 2010,683:97-115.
[28]	FAROOQUI T. A potential link among biogenic aminesbased pesticides, learning and memory, and colony collapse disorder:a unique hypothesis[J]. Neurochemistry International, 2013, 62(1):122-136.
[29]	潘雪莲,杨磊,袁琳琳,等.豆大蓟马钠离子通道基因克隆及分析[J].热带生物学报, 2022, 13(3):249-258.
[30]	HIRATA K, JOURAKU A, KUWAZAKI S, et al. The R81T mutation in the nicotinic acetylcholine receptor of Aphis gossypii is associated with neonicotinoid insecticide resistance with differential effects for cyano-and nitro-substituted neonicotinoids[J]. Pesticide Biochemistry and Physiology, 2017, 143:57-65.
[31]	DEDERER H, BERGER M, MEYER T, et al. Structureactivity relationships of acetylcholine derivatives with Lucilia cuprina nicotinic acetylcholine receptor α1 and α2subunits in chicken β2 subunit hybrid receptors in comparison with chicken nicotinic acetylcholine receptor α4/β2[J]. Insect Molecular Biology, 2013, 22(2):183-198.
[32]	WANG Z, ZHANG R, PEI Y, et al. The knockout of the nicotinic acetylcholine receptor subunit gene α1(nAChRα1) through CRISPR/Cas9 technology exposes its involvement in the resistance of Spodoptera exigua to insecticides[J]. Pesticide Biochemistry and Physiology,2023, 196:105616.
[33]	SHAN T, ZHANG H, CHEN C, et al. Low expression levels of nicotinic acetylcholine receptor subunits Boα1 and Boβ1 are associated with imidacloprid resistance in Bradysia odoriphaga[J]. Pest Management Science, 2020,76(9):3038-3045.
[34]	LEES K, JONES A K, MATSUDA K, et al. Functional characterisation of a nicotinic acetylcholine receptor αsubunit from the brown dog tick, Rhipicephalus sanguineus[J]. International Journal for Parasitology, 2014,44(1):75-81.
[35]	QU Y, CHEN J, LI C, et al. The subunit gene Ldα1 of nicotinic acetylcholine receptors plays important roles in the toxicity of imidacloprid and thiamethoxam against Leptinotarsa decemlineata[J]. Pesticide Biochemistry and Physiology, 2016, 127:51-58.
[36]	LU W, LIU Z, FAN X, et al. Nicotinic acetylcholine receptor modulator insecticides act on diverse receptor subtypes with distinct subunit compositions[J]. PLoS Genetics, 2022, 18(1):e1009920.
[37]	姚香梅.新烟碱类杀虫剂对靶标和非靶标昆虫乙酰胆碱受体毒理学特性研究[D].北京:中国农业科学院,2015.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya, Hainan 572025, China;

2. School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China

Received Date: 2023-12-13

Rev Recd Date: 2024-02-07

Key words:

Abstract: Neonicotinoids that act on insect nicotinic acetylcholine receptors（nAChRs） are common tools for crop pest control, but they have serious negative effects on pollinators such as Apis mellifera. In this study, the nAChR α1 subunit gene was cloned from A. mellifera and systematically analyzed. The interaction mechanism between the nAChR α1 subtype and neonicotinoids（imidacloprid, clothianidin, and dinotefuran） was investigated by using the two-electrode voltage clamp technique. Sequence alignment and phylogenetic tree analysis showed that the nAChR α1 subunit of A. mellifera had typical domain characteristics in the nAChR subunit family genes, and was most distantly related to the α1 subunit of Hemiptera insects and most closely related to the α1 subunit of bees from Hymenoptera. The combination of A. mellifera nAChR α1 subunit and Rattus norvegicus nAChR β2 subunit was co-expressed in Xenopus laevis oocytes. Electrophysiological results showed that imidacloprid had the highest agonistic efficiency and lowest agonistic affinity, that clothianidin had the lowest agonistic efficiency and that dinotefuran had the highest agonistic affinity for Amα1/rβ2 nAChR, indicating that the α1 subunit of A. mellifera nAChRs is one of the targets of neonicotinoids and the sensitivity of Amα1/rβ2 nAChR to different neonicotinoids is different. All these results have important theoretical significance for the development of novel highly selective insecticides for target insects.

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[1]	LIU S, LIU Y, HE F, et al. Enantioselective olfactory effects of the neonicotinoid dinotefuran on honey bees (Apis mellifera L.)[J]. Journal of Agricultural and Food Chemistry, 2019, 67(43):12105-12116.
[2]	KLEIN A M, VAISSIÈRE B E, CANE J H, et al. Importance of pollinators in changing landscapes for world crops[J]. Proceedings Biological Sciences, 2007, 274(1608):303-313.
[3]	GALLAI N, SALLES J M, SETTELE J, et al. Economic valuation of the vulnerability of world agriculture confronted with pollinator decline[J]. Ecological Economics,2009, 68(3):810-821.
[4]	VANENGELSDORP D, UNDERWOOD R, CARON D, et al. Estimate of managed colony losses in the winter of 2006-2007:A report commissioned by the apiary inspectors of America[J]. American Bee Journal, 2007, 147:599-603.
[5]	AURELL D, BRUCKNER S, WILSON M, et al. A national survey of managed honey bee colony losses in the USA:results from the bee informed partnership for 2020-21 and 2021-22[J]. Journal of Apicultural Research, 2024, 63(1):1-14.
[6]	WILLIAMS G R, TARPY D R, VANENGELSDORP D, et al. Colony collapse disorder in context[J]. BioEssays:News and Reviews in Molecular, Cellular and Developmental Biology, 2010, 32(10):845-846.
[7]	VANENGELSDORP D, EVANS J D, SAEGERMAN C, et al. Colony collapse disorder:a descriptive study[J]. PLoS One, 2009, 4(8):e6481.
[8]	CHEN Y P, PETTIS J S, CORONA M, et al. Israeli acute paralysis virus:epidemiology, pathogenesis and implications for honey bee health[J]. PLoS Pathogens, 2014,10(7):e1004261.
[9]	MARTIN S J, HIGHFIELD A C, BRETTELL L, et al.Global honey bee viral landscape altered by a parasitic mite[J]. Science, 2012, 336(6086):1304-1306.
[10]	MESNAGE R, ANTONIOU M N. Ignoring adjuvant toxicity falsifies the safety profile of commercial pesticides[J].Frontiers in Public Health, 2018, 5:361.
[11]	LI G, ZHAO H, LIU Z, et al. The wisdom of honeybee defenses against environmental stresses[J]. Frontiers in Microbiology, 2018, 9:722.
[12]	STRAUB L, WILLIAMS G R, VIDONDO B, et al. Neonicotinoids and ectoparasitic mites synergistically impact honeybees[J]. Scientific Reports, 2019, 9(1):8159.
[13]	FORFERT N, TROXLER A, RETSCHNIG G, et al. Neonicotinoid pesticides can reduce honeybee colony genetic diversity[J]. PLoS One, 2017, 12(10):e0186109.
[14]	张伟,齐素贞,薛晓锋,等.农药对蜜蜂的影响及毒理学机制研究进展[J].农药学学报, 2022, 24(5):1125-1138.
[15]	LIU J, LI Y, ZHANG Z, et al. Low concentration of quercetin reduces the lethal and sublethal effects of imidacloprid on Apis cerana (Hymenoptera:Apidae)[J]. Journal of Economic Entomology, 2021, 114(3):1053-1064.
[16]	TOSI S, NIEH J C. A common neonicotinoid pesticide,thiamethoxam, alters honey bee activity, motor functions,and movement to light[J]. Scientific Reports, 2017, 7(1):15132.
[17]	TSVETKOV N, SAMSON-ROBERT O, SOOD K, et al.Chronic exposure to neonicotinoids reduces honey bee health near corn crops[J]. Science, 2017, 356(6345):1395-1397.
[18]	IHARA M, MATSUDA K. Neonicotinoids:molecular mechanisms of action, insights into resistance and impact on pollinators[J]. Current Opinion in Insect Science,2018, 30:86-92.
[19]	CHANGEUX J P. The nicotinic acetylcholine receptor:the founding father of the pentameric ligand-gated ion channel superfamily[J]. The Journal of Biological Chemistry, 2012, 287(48):40207-40215.
[20]	IHARA M, HIKIDA M, MATSUSHITA H, et al. Loops D, E and G in the Drosophila Dα1 subunit contribute to high neonicotinoid sensitivity of Dα1-chicken β2 nicotinic acetylcholine receptor[J]. British Journal of Pharmacology, 2018, 175(11):1999-2012.
[21]	李旭扬,陈舒苗,于津鹏,等. α6/α3β4烟碱型乙酰胆碱受体β4亚基双点突变体的制备及其功能研究[J].热带生物学报, 2021, 12(4):403-411.
[22]	马倩芸,江涛,于日磊.烟碱乙酰胆碱受体结构的研究进展[J].中国海洋药物, 2018, 37(2):97-102.
[23]	ZHOU Y, HAN Q, FENG K, et al. New binding sites of nicotinic acetylcholine receptors from Myzus persicae[J].Entomologia Generalis, 2023, 43(2):461-470.
[24]	LIU Z, WILLIAMSON M S, LANSDELL S J, et al. A nicotinic acetylcholine receptor mutation (Y151S) causes reduced agonist potency to a range of neonicotinoid insecticides[J]. Journal of Neurochemistry, 2006, 99(4):1273-1281.
[25]	DEDERER H, WERR M, ILG T. Differential sensitivity of Ctenocephalides felis and Drosophila melanogaster nicotinic acetylcholine receptor α1 and α2 subunits in recombinant hybrid receptors to nicotinoids and neonicotinoid insecticides[J]. Insect Biochemistry and Molecular Biology, 2011, 41(1):51-61.
[26]	WANG Z M, LI S, SHI C C, et al. The actions of neonicotinoid insecticides on nicotinic acetylcholine subunits Ldα1 and Ldα8 of Leptinotarsa decemlineata and assembled receptors[J]. Insect Science, 2022, 29(5):1387-1400.
[27]	GAUTHIER M. State of the art on insect nicotinic acetylcholine receptor function in learning and memory[J].Advances in Experimental Medicine and Biology, 2010,683:97-115.
[28]	FAROOQUI T. A potential link among biogenic aminesbased pesticides, learning and memory, and colony collapse disorder:a unique hypothesis[J]. Neurochemistry International, 2013, 62(1):122-136.
[29]	潘雪莲,杨磊,袁琳琳,等.豆大蓟马钠离子通道基因克隆及分析[J].热带生物学报, 2022, 13(3):249-258.
[30]	HIRATA K, JOURAKU A, KUWAZAKI S, et al. The R81T mutation in the nicotinic acetylcholine receptor of Aphis gossypii is associated with neonicotinoid insecticide resistance with differential effects for cyano-and nitro-substituted neonicotinoids[J]. Pesticide Biochemistry and Physiology, 2017, 143:57-65.
[31]	DEDERER H, BERGER M, MEYER T, et al. Structureactivity relationships of acetylcholine derivatives with Lucilia cuprina nicotinic acetylcholine receptor α1 and α2subunits in chicken β2 subunit hybrid receptors in comparison with chicken nicotinic acetylcholine receptor α4/β2[J]. Insect Molecular Biology, 2013, 22(2):183-198.
[32]	WANG Z, ZHANG R, PEI Y, et al. The knockout of the nicotinic acetylcholine receptor subunit gene α1(nAChRα1) through CRISPR/Cas9 technology exposes its involvement in the resistance of Spodoptera exigua to insecticides[J]. Pesticide Biochemistry and Physiology,2023, 196:105616.
[33]	SHAN T, ZHANG H, CHEN C, et al. Low expression levels of nicotinic acetylcholine receptor subunits Boα1 and Boβ1 are associated with imidacloprid resistance in Bradysia odoriphaga[J]. Pest Management Science, 2020,76(9):3038-3045.
[34]	LEES K, JONES A K, MATSUDA K, et al. Functional characterisation of a nicotinic acetylcholine receptor αsubunit from the brown dog tick, Rhipicephalus sanguineus[J]. International Journal for Parasitology, 2014,44(1):75-81.
[35]	QU Y, CHEN J, LI C, et al. The subunit gene Ldα1 of nicotinic acetylcholine receptors plays important roles in the toxicity of imidacloprid and thiamethoxam against Leptinotarsa decemlineata[J]. Pesticide Biochemistry and Physiology, 2016, 127:51-58.
[36]	LU W, LIU Z, FAN X, et al. Nicotinic acetylcholine receptor modulator insecticides act on diverse receptor subtypes with distinct subunit compositions[J]. PLoS Genetics, 2022, 18(1):e1009920.
[37]	姚香梅.新烟碱类杀虫剂对靶标和非靶标昆虫乙酰胆碱受体毒理学特性研究[D].北京:中国农业科学院,2015.

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Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

doi: 10.15886/j.cnki.rdswxb.20230140

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References

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通讯作者: 陈斌, bchen63@163.com

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Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

doi: 10.15886/j.cnki.rdswxb.20230140

1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya, Hainan 572025, China;

2. School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China

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Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

doi: 10.15886/j.cnki.rdswxb.20230140

Abstract

References

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通讯作者: 陈斌, bchen63@163.com

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Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

doi: 10.15886/j.cnki.rdswxb.20230140

1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya, Hainan 572025, China; 2. School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China

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1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya, Hainan 572025, China;

2. School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China