GABA responses and their partial occlusion by glycine in cultured rat medullary neurons

Abstract

Whole-cell current responses to bath application of GABA and glycine were studied in medullary neurons cultured from embryonic rats. Two current components were seen in the responses to bath application of GABA, one component which desensitized and another which did not. These two current components have different dose-response characteristics for GABA, with the nondesensitizing component being activated more effectively and reaching its peak amplitude at lower agonist concentrations than the desensitizing one. The agonist concentrations producing half of the maximum responses are 2.8 +/- 0.3 (+/- S.E.M., n = 9) and 14.7 +/- 2.7 (n = 5) microM for the nondesensitizing and desensitizing components, respectively. The two current components for GABA are differentially affected by the antagonists, picrotoxin and bicuculline. The antagonist concentrations which block 50% of the control desensitizing and nondesensitizing responses to GABA are 33 and 320 microM for picrotoxin, and 3 and 50 microM for bicuculline, respectively. Thus, the characteristics of the GABA responses are analogous to those described previously for glycine in that there are two components which are differentially sensitive to agonist concentration [Lewis et al. (1991) J. Neurophysiol, 40, 1178-1187]. We now find there is occlusion between the responses to GABA and glycine, indicating that they share a population of receptors or channels. The occlusion was incomplete (< 80%) in half of the cells, suggesting that both agonists also activate unique receptors. Furthermore, the current responses to 35 microM GABA are blocked by the glycinergic antagonist, strychnine, with half-maximal blocking concentrations equal to 2 and 30 microM for the desensitizing and nondesensitizing components, respectively. This strychnine sensitivity is less than that for the glycine receptor. At the same time, the current responses to 100 microM glycine are sensitive to the GABAergic antagonists, picrotoxin and bicuculline. The half-maximal blocking concentrations are 36 and 120 microM picrotoxin, and 120 and 500 microM bicuculline, for the desensitizing and nondesensitizing components of the glycine response, respectively. Consequently, these results suggest that these cultured cells have at least three types of inhibitory receptors: glycine receptors, GABA receptors and GABA/glycine receptors, with all three receptors sensitive to block by strychnine, bicuculline and picrotoxin. The GABA/glycine receptor may be an immature form of the inhibitory receptor. Alternatively, some GABA and glycine receptors may have common ionophores.