Drosophila nicotinic acetylcholine receptors and their interactions with insecticidal peptide toxins
Drosophila nicotinic acetylcholine receptors and their interactions with insecticidal peptide toxins
Abstract Drosophila nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that represent a target for insecticides. Peptide neurotoxins are known to block nAChRs by binding to their target subunits, however, a better understanding of receptor subunit composition is needed for effective design of insecticides. To facilitate the analysis of nAChRs we used a CRISPR/Cas9 strategy to generate null alleles for all ten nAChR subunit genes in a common genetic background. We studied interactions of nAChR subunits with peptide neurotoxins by larval injections and styrene maleic acid lipid particles (SMALPs) pull-down assays. For the null alleles we determined the effects of α-Bungarotoxin (α-Btx) and ω-Hexatoxin-Hv1a (Hv1a) administration, identifying potential receptor subunits implicated in the binding of these toxins. We employed pull-down assays to confirm α-Btx interactions with the Dα5, Dα6, Dα7 subunits. Finally, we report the localization of fluorescent tagged endogenous Dα6 during nervous system development. Taken together this study elucidates native Drosophila nAChR subunit interactions with insecticidal peptide toxins and provides a resource for the in vivo analysis of insect nAChRs.
Lilley Kathryn S、Dirnberger Benedict、Giachello Carlo N G、Minde David-Paul、Deery Michael J、M¨1ller Karin H、Korona Dagmara、Russell Steven、Johnson Glynnis、Queiroz Rayner M L、Earley Fergus G、Firth Lucy C
Cambridge Centre for Proteomics, Department of Biochemistry, Gleeson Building, University of CambridgeDepartment of Genetics, University of Cambridge||2Cambridge Centre for Proteomics, Department of Biochemistry, Gleeson Building, University of Cambridge||Syngenta, Jealott?ˉs Hill International Research CentreSyngenta, Jealott?ˉs Hill International Research Centre2Cambridge Centre for Proteomics, Department of Biochemistry, Gleeson Building, University of Cambridge2Cambridge Centre for Proteomics, Department of Biochemistry, Gleeson Building, University of CambridgeCambridge Advanced Imaging Centre, Department of Physiology, Development and Neuroscience/Anatomy Building, University of CambridgeDepartment of Genetics, University of CambridgeDepartment of Genetics, University of CambridgeDepartment of Genetics, University of Cambridge2Cambridge Centre for Proteomics, Department of Biochemistry, Gleeson Building, University of CambridgeSyngenta, Jealott?ˉs Hill International Research CentreSyngenta, Jealott?ˉs Hill International Research Centre
昆虫学分子生物学生理学
Drosophila nAChRsinsecticidal toxinsnAChR null allelesSMALPs pull-downs
Lilley Kathryn S,Dirnberger Benedict,Giachello Carlo N G,Minde David-Paul,Deery Michael J,M¨1ller Karin H,Korona Dagmara,Russell Steven,Johnson Glynnis,Queiroz Rayner M L,Earley Fergus G,Firth Lucy C.Drosophila nicotinic acetylcholine receptors and their interactions with insecticidal peptide toxins[EB/OL].(2025-03-28)[2025-08-02].https://www.biorxiv.org/content/10.1101/2021.08.13.456240.点此复制
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