Identification and function of glycine receptors in cultured cerebellar granule cells

Poly(A)+ mRNA was isolated from cultured mouse cerebellar granule cells and injected into Xenopus oocytes. This led to the expression of receptors that evoked large membrane currents in response to glycine. Current-responses were also obtained after application of beta-alanine and taurine, but these were very low relative to that of glycine (maximal beta-alanine and taurine responses were 8 and 3% of that of glycine, respectively). The role of glycine receptors on K(+)-evoked transmitter release in cultured cerebellar granule cells was also assayed. Release of preloaded D-[3H]aspartate evoked by 40 mM K+ was dose dependently inhibited by glycine, and the concentration producing half-maximal inhibition was 50 microM. Taurine, beta-alanine, and the specific GABAA receptor agonist isoguvacine also inhibited K(+)-evoked release, and the maximal inhibition was similar for all agonists (approximately 40%). The EC50 value was 200 microM for taurine, 70 microM for beta-alanine, and 4 microM for isoguvacine. Bicuculline (150 microM) antagonized the inhibitory effect of isoguvacine (150 microM) but not that of glycine (1 mM). In contrast, strychnine (20 microM) antagonized the inhibitory effect of glycine (1 mM) but not that of isoguvacine (150 microM). The pharmacology of the responses to beta-alanine and taurine showed that these agonists activate both glycine and GABAA receptors. The results indicate that cultured cerebellar granule cells translate the gene for the glycine receptor and that activation of glycine receptors produces neuronal inhibition.