Abstract

Poly(A)+ RNA from rat cerebral cortex expresses gamma-aminobutyric acid (GABA)-activated membrane current responses in Xenopus oocytes, mediated by GABAA receptors (IG-Actx). In contrast, RNA from bovine retina expresses GABA responses composed of two pharmacologically distinct Cl- currents, one mediated by GABAA receptors (IG-Aret) and the other by atypical GABA receptors that are resistant to bicuculline and are not activated by baclofen (IG-BR). The pharmacology of the bicuculline/baclofen-insensitive GABA receptors was further investigated by comparing actions of hexachlorocyclohexane (HCH) enantiomers on GABA-activated membrane currents expressed in oocytes by brain and retina RNA. gamma-HCH (lindane) was a potent inhibitor of IG-Actx, with suppression of currents detectable at concentrations as low as 50 nM. The IC50 for gamma-HCH, calculated from inhibitory effects on maximum IG-Actx (current elicited by 3 mM GABA), was 7.3 +/- 3 microM. Inhibitory effects of gamma-HCH on IG-Aret were qualitatively similar to those described for IG-Actx. In contrast, alpha-HCH and delta-HCH induced clear positive modulation of IG-Actx elicited by low (e.g., 10 microM) concentrations of GABA. Thresholds for the modulatory effects of alpha-HCH and delta-HCH were between 100 and 300 nM, with maximum levels of potentiation (5-7-fold) between 20-50 microM. Potentiation of IG-Actx by alpha- and delta-HCH was reversible and largely insensitive to the benzodiazepine antagonist flumazenil (1 microM). Assays on maximum IG-Actx indicated that alpha-HCH (10-100 microM) caused only marginal reductions in response (less than or equal to 15%), whereas delta-HCH had stronger inhibitory effects (IC50, 20-30 microM). At concentrations between 0.1 and 50 microM, beta-HCH induced only 10-25% facilitation of IG-Actx elicited by 10 microM GABA and had no clear effects on maximum responses. IG-BR was also potently inhibited by gamma-HCH. Thresholds for detecting reductions in current were approximately 20 nM, and the IC50 calculated from effects on maximum responses was 5.8 +/- 2 microM. However, neither alpha-HCH nor delta-HCH (1-100 microM) induced any potentiation of IG-BR. alpha-HCH had some weak inhibitory effects that were largely surmountable, whereas delta-HCH and beta-HCH were essentially inactive. These experiments raise the possibility that alpha- and delta-HCH constitute a novel class of GABAA receptor modulators, which might prove to be useful for investigating the mechanisms underlying regulation of GABAA receptors.(ABSTRACT TRUNCATED AT 400 WORDS)