Wild-type and insecticide-resistant homo-oligomeric GABA receptors of Drosophila melanogaster stably expressed in a Drosophila cell line.
RDL is an ionotropic GABA receptor subunit, a product of the Rdl gene, originally identified in the Maryland strain of Drosophila melanogaster. Here, we report the generation of a Drosophila melanogaster cell line (S2-RDLA302S) stably expressing a mutated, dieldrin-resistant (A302S) form of RDL. The properties of this dieldrin-resistant, homo-oligomeric receptor have been compared with those of the stably expressed, wild-type form (S2-RDL). Using these stable lines, a striking reduction in sensitivity to both picrotoxinin and dieldrin was observed for responses to GABA of S2-RDLA302S compared to S2-RDL. To determine if these stable insect cell lines generate results similar to those obtained by transient expression in Xenopus laevis oocytes, we have examined the actions of two widely used convulsants, EBOB and TBPS, and a recently developed convulsant BIDN, on RDL-mediated GABA responses in the two expression systems. In both oocytes and S2 cells, the three convulsants suppressed the amplitude of responses to GABA. Thus, in accord with earlier work on agonist and allosteric sites, the S2-RDL cell line is found to yield similar pharmacological results to those obtained in transient expression studies. Stable cell lines are now available expressing susceptible and resistant forms of an ionotropic receptor by GABAergic insecticides.
Buckingham
S D
SD
Babraham Institute Laboratory of Molecular Signalling, Department of Zoology, University of Cambridge, UK.
Matsuda
K
K
Hosie
A M
AM
Baylis
H A
HA
Squire
M D
MD
Lansdell
S J
SJ
Millar
N S
NS
Sattelle
B
B
D000818:Animals; D002460:Cell Line; D004026:Dieldrin / Q000494:pharmacology; D004331:Drosophila melanogaster; D004594:Electrophysiology; D018755:GABA Agonists / Q000494:pharmacology; D007305:Insecticide Resistance / Q000235:genetics*; D007306:Insecticides / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D018079:Receptors, GABA / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D014162:Transfection; D014982:Xenopus laevis
D018755:GABA Agonists; D007306:Insecticides; D018079:Receptors, GABA; D004026:Dieldrin