| 10212398 |
Wolff MA, Wingate VP: Characterization and comparative pharmacological studies of a functional gamma-aminobutyric acid (GABA) receptor cloned from the tobacco budworm, Heliothis virescens (Noctuidae:Lepidoptera). Invert Neurosci. 1998 Mar;3(4):305-15.
This paper reports the functional expression and pharmacological characterization of a full length complementary deoxyribonucleic acid (cDNA) (pIVY12) cloned from a Heliothis virescens fertilized egg cDNA library that encodes for a gamma-aminobutyric acid (GABA) receptor subunit (HVRDL-Ser 285). Two electrode voltage clamp recordings of Xenopus oocytes expressing the HVRDL GABA-gated chloride channel revealed robust chloride ion conductance in response to GABA and the GABAA receptor agonist, muscimol. Baclofen, a GABAB agonist had no effect. Phenobarbital showed a positive dose-dependent allosteric modulatory effect, whereas the benzodiazepine, flunitrazepam, had no effect. Chloride conductance was depressed by the novel insecticide, fipronil ((+/-)-5-amino-1-(2,6 dichloro-alpha, alpha, alpha-trifluoro-p-tolyl)-4-trifluoromethyl-sulfinylpyrazole-3-carb onitrile) and the GABAA antagonist, picrotoxinin. The HVRDL GABA receptor was insensitive to blockage by dieldrin and the GABAA antagonist, bicuculline. The comparative actions of fipronil, picrotoxinin and dieldrin were examined on oocytes expressing the H. virescens wild-type (HVRDL-Ser 285), the site-directed mutant (HVRDL-Ala 285), the Drosophila melanogaster Rdl wild-type (DMRDL-Ala 302) and the Rdl dieldrin resistant (DMRDL-Ser 302) homo-oligomeric GABA receptors. HVRDL-Ala 285 was 15-fold more sensitive to blockage by fipronil than HVRDL-Ser 285. DMRDL-Ala 302 and DMRDL-Ser-302 showed a similar level of sensitivity to blockage by fipronil. HVRDL-Ser 285 and DMRDL-Ser 302 exhibited a similar level of insensitivity to picrotoxinin. HVRDL-Ala 285 and DMRDL-Ala 302 showed a similar range of picrotoxinin sensitivity. DMRDL-Ala 302 and HVRDL-Ala 285 showed some sensitivity to blockage by dieldrin. Fipronil sensitivity was significantly altered by the serine to alanine mutation at position 285 in the M2 region of the HVRDL subunit, whereas no difference was observed between the DMRDL-Ser 302 and DMRDL-Ala 302 receptors. |
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