Two components of metabotropic glutamate responses in acutely dissociated CA3 pyramidal neurons of the rat.

The excitatory and inhibitory actions of metabotropic glutamate receptor (mGluR) agonists were investigated in acutely dissociated rat hippocampal CA3 pyramidal neurons, using the conventional whole-cell and nystatin-perforated patch recording configurations under the voltage-clamp condition. With the conventional whole-cell recording, glutamate (Glu) and quisqualic acid (QA) induced only ionotropic inward currents accompanied by increased membrane conductance at a holding potential (VH) of -45 mV. The response was reversibly blocked in the presence of D-2-amino-5-phosphonopentanoic acid (D-AP5) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), the antagonists of N-methyl-D-aspartate (NMDA) receptor and non-NMDA receptor, respectively. With nystatin-perforated patch recording, mGlu responses insensitive to both D-AP5 and CNQX were observed. Fifty-five % of the cells responded by a slow inward current accompanied by conductance decrease (ImGlui) at a VH of -44 mV. One % of the neurons showed an outward current with conductance increase (ImGluo), and 34% of the neurons showed ImGluo followed by ImGlui. The onset of ImGluo occurred approximately 900 ms after the response to 30 mM K+. The time to peak of ImGluo were 32- to 79-times longer than those of ionotropic responses. ImGlui appeared at lower concentrations than ionotropic Glu responses, whereas ImGluo appeared at similar concentrations as ionotropic responses. The rank order of affinity was QA > Glu > (+/-)-1-aminocyclopentane-trans-1, 3-dicarboxylic acid (tACPD) for both ImGlui and ImGluo. Half-maximal effective concentrations (EC50) and the threshold concentrations for the three agonists were four- to tenfold lower for ImGlui than for ImGluo. The current-voltage relationship showed that the reversal potentials of ImGlui and ImGluo shifted 55 and 59 mV, respectively, for a tenfold change in extracellular K+ concentration, indicating that K+ is the charge carrier of both mGlu responses. During ImGlui, both the leakage current and muscarine-sensitive voltage-dependent K+ current (M current) were suppressed. ImGluo induced by 10(-4) M tACPD was abolished by 3.10(-7) M charybdotoxin and 10(-6) M ryanodine. These results show that there are two components of mGlu responses in CA3 pyramidal neurons and that ImGlui and ImGluo show different pharmacological properties.