title: Mosquitocidal properties of IgG targeting the glutamate-gated chloride channel in three mosquito disease vectors (Diptera: Culicidae).
abstract: The glutamate-gated chloride channel (GluCl) is a highly sensitive insecticide target of the avermectin class of insecticides. As an alternative to using chemical insecticides to kill mosquitoes, we tested the effects of purified immunoglobulin G (IgG) targeting the extracellular domain of GluCl from Anopheles gambiae (AgGluCl) on the survivorship of three key mosquito disease vectors: Anopheles gambiae s.s., Aedes aegypti and Culex tarsalis. When administered through a single blood meal, anti-AgGluCl IgG reduced the survivorship of A. gambiae in a dose-dependent manner (LC50: 2.82 mg ml(-1), range 2.68-2.96 mg ml(-1)) but not A. aegypti or C. tarsalis. We previously demonstrated that AgGluCl is only located in tissues of the head and thorax of A. gambiae. To verify that AgGluCl IgG is affecting target antigens found outside the midgut, we injected it directly into the hemocoel via intrathoracic injection. A single, physiologically relevant concentration of anti-AgGluCl IgG injected into the hemocoel equally reduced mosquito survivorship of all three species. To test whether anti-AgGluCl IgG was entering the hemocoel of each of these mosquitoes, we fed mosquitoes a blood meal containing anti-AgGluCl IgG and subsequently extracted their hemolymph. We only detected IgG in the hemolymph of A. gambiae, suggesting that resistance of A. aegypti and C. tarsalis to anti-AgGluCl IgG found in blood meals is due to deficient IgG translocation across the midgut. We predicted that anti-AgGluCl IgG's mode of action is by antagonizing GluCl activity. To test this hypothesis, we fed A. gambiae blood meals containing anti-AgGluCl IgG and the GluCl agonist ivermectin (IVM). Anti-AgGluCl IgG attenuated the mosquitocidal effects of IVM, suggesting that anti-AgGluCl IgG antagonizes IVM-induced activation of GluCl. Lastly, we stained adult, female A. aegypti and C. tarsalis for GluCl expression. Neuronal GluCl expression in these mosquitoes was similar to previously reported A. gambiae GluCl expression; however, we also discovered GluCl staining on the basolateral surface of their midgut epithelial cells, suggesting important physiological differences in Culicine and Anopheline mosquitoes.
authors.affiliation: Department of Biomedical Sciences, Colorado State University, 1617 Campus Delivery, Fort Collins, CO 80523-1617, USA Jacob.I.Meyers@gmail.com.
authors.forename: Jacob I
authors.initials: JI
authors.lastname: Meyers
authors.affiliation: Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, 1692 Campus Delivery, Fort Collins, CO 80523-1692, USA.
authors.forename: Meg
authors.initials: M
authors.lastname: Gray
authors.affiliation: Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, 1692 Campus Delivery, Fort Collins, CO 80523-1692, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Foy
mesh_terms: D000330:Aedes / Q000187:drug effects* / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000187:drug effects* / Q000502:physiology; D000941:Antigens / Q000276:immunology; D018118:Chloride Channels / Q000276:immunology / Q000378:metabolism*; D003465:Culex / Q000187:drug effects* / Q000502:physiology; D005260:Female; D006458:Hemolymph / Q000276:immunology; D007074:Immunoglobulin G / Q000494:pharmacology*; D007300:Insect Control; D007303:Insect Vectors; D007306:Insecticides / Q000494:pharmacology*; D007559:Ivermectin / Q000031:analogs & derivatives / Q000494:pharmacology; D008969:Molecular Sequence Data; D013045:Species Specificity
chemical_list: D000941:Antigens; D018118:Chloride Channels; D007074:Immunoglobulin G; D007306:Insecticides; C109777:glutamate-gated chloride channels; D007559:Ivermectin; C019264:avermectin
keywords: Aedes aegypti; Anopheles gambiae; Culex tarsalis; GluCl; Mosquitocidal antibody