title: Insecticide sensitivity of native chloride and sodium channels in a mosquito cell line.
abstract: The aim of this study was to investigate the utility of cultured Anopheles gambiae Sua1B cells for insecticide screening applications without genetic engineering or other treatments. Sua1B cells were exposed to the known insecticidal compounds lindane and DIDS, which inhibited cell growth at micromolar concentrations. In patch clamp studies, DIDS produced partial inhibition (69%) of chloride current amplitudes, and an IC50 of 5.1μM was determined for Sua1B cells. A sub-set of chloride currents showed no response to DIDS; however, inhibition (64%) of these currents was achieved using a low chloride saline solution, confirming their identity as chloride channels. In contrast, lindane increased chloride current amplitude (EC50=116nM), which was reversed when cells were bathed in calcium-free extracellular solution. Voltage-sensitive chloride channels were also inhibited by the presence of fenvalerate, a type 2 pyrethroid, but not significantly blocked by type 1 allethrin, an effect not previously shown in insects. Although no evidence of fast inward currents typical of sodium channels was observed, studies with fenvalerate in combination with veratridine, a sodium channel activator, revealed complete inhibition of cell growth that was best fit by a two-site binding model. The high potency effect was completely inhibited in the presence of tetrodotoxin, a specific sodium channel blocker, suggesting the presence of some type of sodium channel. Thus, Sua1B cells express native insect ion channels with potential utility for insecticide screening.
authors.affiliation: Emerging Pathogens Institute, Department of Entomology and Nematology, University of Florida, Gainesville, FL 32601, United States; Virginia Polytechnic Institute and State University, Department of Entomology, Blacksburg, VA 24061, United States.
authors.forename: Lacey J
authors.initials: LJ
authors.lastname: Jenson
authors.affiliation: Virginia Polytechnic Institute and State University, Department of Entomology, Blacksburg, VA 24061, United States.
authors.forename: Troy D
authors.initials: TD
authors.lastname: Anderson
authors.affiliation: Emerging Pathogens Institute, Department of Entomology and Nematology, University of Florida, Gainesville, FL 32601, United States. Electronic address: jbquist@epi.ufl.edu.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Bloomquist
mesh_terms: D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D000852:Anopheles / Q000187:drug effects*; D002460:Cell Line; D018118:Chloride Channels / Q000187:drug effects*; D001556:Hexachlorocyclohexane / Q000494:pharmacology; D007306:Insecticides / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D015222:Sodium Channels / Q000187:drug effects*
chemical_list: D018118:Chloride Channels; D007306:Insecticides; D015222:Sodium Channels; D001556:Hexachlorocyclohexane; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
keywords: DIDS; Fenvalerate; Lindane; Sua1B; Tetrodotoxin; Veratridine