Research ReportEffects of inhibitory amino acids on expression of GABAA Rα and glycine Rα1 in hypoxic rat cortical neurons during development

Recent studies suggest that GABA and glycine are protective to mature but toxic to immature cortical neurons during prolonged hypoxia. Since the action of these inhibitory amino acids is mediated by GABA and glycine receptors, the expression of these receptors is a critical factor in determining neuronal response to GABAA and glycine in hypoxia. Therefore, we asked whether in rat cortical neurons, 1) hypoxia alters the expression of the GABA and glycine receptors; 2) inhibitory amino acids change the course of GABA and glycine receptor expression; and 3) there are any differences between the immature and mature neurons. In cultured rat cortical neurons from day 4 (four days in vitro or DIV 4) to day 20 (DIV 20), we observed that 1) GABAARα and GlyRα1 underwent differential changes in expression during the development in vitro; 2) hypoxia for 3 days decreased GABAARα and GlyRα1 density in the neurons in-between DIV 4 and DIV 20, but did not induce a major change in immature (DIV 4) and mature (DIV 20) neurons; 3) during normoxia GABA, glycine and taurine decreased GABAARα and GlyRα1 density in the immature neurons, but had a tendency to increase the density in the mature neurons, except for taurine; 4) under hypoxia, all these amino acids decreased GABAARα and GlyRα1 density in most groups of the immature neurons with a slight effect on the mature neurons; and 5) δ-opioid receptor activation with DADLE increased GABAARα and GlyRα1 density in both the immature and mature neurons under normoxia and in the mature neurons under hypoxic condition. These data suggest that inhibitory amino acids differentially regulate the expression of GABAA and glycine receptors in rat cortical neurons under normoxic and hypoxic conditions with major differences between the immature and mature neurons.

► The developing profiles of GABAARα and GlyRα1 vary greatly in cortical neurons. ► GABA and glycine decrease GABAARα and GlyRα1 in the immature, but not mature, neurons. ► 3-day hypoxia significantly reduces GABAARα and GlyRα1 density in developing neurons. ► GABA, glycine and taurine reduce GABAARα and GlyRα1 in the immature neurons in hypoxia. ► DOR activation increases GABAARα and GlyRα1 in both immature and mature neurons.