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摘要
Due to the great abundance within insect central nervous system (CNS), nicotinic acetylcholine receptors (nAChRs) play key roles in insect CNS, which makes it to be the targets of several classes of insecticides, such as neonicotinoids. Insect nAChRs are pentameric complexes consisting of five subunits, and a dozen subunits in one insect species can theoretically comprise diverse nAChRs. The alternative splicing in insect nAChR subunits may increase the diversity of insect nAChRs. In the oriental migratory locust (Locusta migratoria manilensis Meyen), a model insect species with agricultural importance, the alternative splicing was found in six alpha subunits among nine alpha and two beta subunits, such as missing conserved residues in Loop D from Loc alpha l, Loc alpha 6 and Loc alpha 9, a 34-residue insertion in Loc alpha 8 cytoplasmic loop, and truncated transcripts for Loc alpha 4, Loc alpha 7 and Loc alpha 9. Hybrid nAChRs were successfully constructed in Xenopus oocytes through co-expression with rat beta 2 and one a subunit from L. migratoria, which included Loc alpha l, Loc alpha l, Loc alpha 3, Loc alpha 4, Loc alpha 5, Loc alpha 8 and Loc alpha 9. Influences of alternative splicing in Loc alpha l, Loc alpha 8 and Loc alpha 9 on acetylcholine potency were tested on hybrid nAChRs. The alternative splicing in Loc alpha l and Loc alpha 9 could increase acetylcholine sensitivities on recombinant receptors, while the splicing in Loc alpha 8 showed significant influences on the current amplitudes of oocytes. The results revealed that the alternative splicing at or close to the ligand-binding sites, as well as at cytoplasmic regions away from the ligandbinding sites, in insect nAChR subunits would change the agonist potencies on the receptors, which consequently increased nAChR diversity in functional and pharmacological properties.
- 出版日期2017-1-18
- 单位南京农业大学
