Toxicology Letters 122 (2001) 215 – 222 GABA receptor subunit composition relative to insecticide potency and selectivity Gurpreet S. Ratra, John E. Casida * Enironmental Chemistry and Toxicology Laboratory, Department of Enironmental Science, Policy and Management, 115 Wellman Hall, Uniersity of California, Berkeley, CA 94720 -3112, USA Received 14 March 2001; received in revised form 20 April 2001; accepted 1 May 2001 Abstract Three observations on the 4-[ 3 H]propyl-4-ethynylbicycloorthobenzoate ([ 3 H]EBOB) binding site in the -aminobu- tyric acid (GABA) receptor indicate the specific target for insecticide action in human brain and a possible mechanism for selectivity. First, from published data, -endosulfan, lindane and fipronil compete for the [ 3 H]EBOB binding site with affinities of 0.3 – 7 nM in both human recombinant homooligomeric 3 receptors and housefly head membranes. Second, from structure-activity studies, including new data, GABAergic insecticide binding potency on the pentameric receptor formed from the 3 subunit correlates well with that on the housefly receptor (r =0.88, n =20). This conserved inhibitor specificity is consistent with known sequence homologies in the housefly GABA receptor and the human GABA A receptor 3 subunit. Third, as mostly new findings, various combinations of 1, 6, and 2 subunits coexpressed with a 1 or 3 subunit confer differential insecticide binding sensitivity, particularly to fipronil, indicating that subunit composition is a major factor in insecticide selectivity. © 2001 Elsevier Science Ireland Ltd. All rights reserved. Keywords: -Aminobutryic acid; Binding site specificity; [ 3 H]EBOB; -Endosulfan; Fipronil; GABA receptor; GABA receptor subunits; Insecticide; Lindane; Subunit composition www.elsevier.com/locate/toxlet 1. Introduction Insecticides selectively toxic to insects compared with humans play an essential role in crop protec- tion and public health. One of their major targets is the -aminobutyric acid (GABA)-gated chloride channel. There are three major classes of GABAergic insecticides exemplified by -endosul- fan, lindane and fipronil (Fig. 1) (Casida, 1993; Bloomquist, 2001). Selective toxicity is conferred in part by target-site specificity between mammals and insects (Cole and Casida, 1992; Hawkinson and Casida, 1993; Casida and Pulman, 1994). The mammalian GABA A pentameric receptor consists primarily of heterooligomeric assemblies of (1–6), (1–4), and (1 – 4) subunits. Recom- binant subunits can be expressed individually or in various combinations (some of which form * Corresponding author. Tel.: +1-510-642-5424; fax: +1- 510-642-6497. E-mail address: ectl@nature.berkeley.edu (J.E. Casida). 0378-4274/01/$ - see front matter © 2001 Elsevier Science Ireland Ltd. All rights reserved. PII:S0378-4274(01)00366-6