Benzodiazepine interactions with GABAA receptors on chick ciliary ganglion neurons

AE McEachern and DK Berg

Department of Biology, University of California, San Diego, La Jolla 92093.

gamma-Aminobutyric acidA (GABAA) receptors on chick ciliary ganglion neurons can be modulated by benzodiazepines and identified by radiolabeled benzodiazepine binding. Enhancement of submaximal GABA responses by benzodiazepines was demonstrated using a multibarrel pipette to construct complete benzodiazepine dose-response curves for single cells in culture. EC50 values of 22 +/- 5 nM, 1.1 +/- 0.3 microM, and 4.6 +/- 0.5 microM were obtained for flunitrazepam, clonazepam, and chlordiazepoxide, respectively. Chlordiazepoxide shifted the GABA dose-response curve to lower GABA concentrations without increasing the maximal response to GABA, demonstrating that benzodiazepines enhance the GABA response by increasing the receptor affinity for GABA. The imidazodiazepine Ro15-1788 potentiated the GABA response with an EC50 of 250 +/- 70 nM, and Ro5-4864 (chlorodiazepam) partially blocked the GABA response both in the presence and absence of chlordiazepoxide. Scatchard analysis of data from binding studies with [3H]flunitrazepam to ganglion membrane homogenates was consistent with the presence of a single class of high affinity sites with a KD of 34 +/- 6 nM and a Bmax of 145 +/- 26 fmol/mg of protein. Several lines of evidence indicated that the sites were associated with GABAA receptors. The KD of [3H]flunitrazepam binding was similar to the EC50 for flunitrazepam modulation of the GABA response. The level of [3H]flunitrazepam binding was enhanced approximately 50% over control levels by GABA. The binding was decreased both by clonazepam and by Ro5- 4864 at concentrations similar to those required for the compounds to modulate the GABA response. These studies demonstrate that ciliary ganglion GABAA receptors are similar in major respects to GABAA receptors in the central nervous system but may differ in minor pharmacological properties.

Volume 34, Issue 2, pp. 129-135, 08/01/1988
Copyright © 1988 by American Society for Pharmacology and Experimental Therapeutics