Effect of the infusion of the GABA-A receptor agonist, muscimol, on the role of the entorhinal cortex, amygdala, and hippocampus in memory processes.

Rats were bilaterally implanted with cannulae in the entorhinal cortex, amygdala, and hippocampus; after recovery, they were trained in a step-down inhibitory avoidance task and tested for retention 24 h later. Muscimol (0.03 microgram) or D-amino-5-phosphonovalerate (5.0 micrograms) infused in the entorhinal cortex 20 min prior to training inhibited the amnestic effect of the same dose of muscimol infused into this area 100 min after training. Thus, memory-relevant information must be processed by the entorhinal cortex at the time of training in order that this cortex may play a late post-training role in memory processing. Pretraining intraentorhinal muscimol administration did not affect the amnestic effect of the post-training infusion of muscimol into the amygdala and hippocampus, or the inhibition of memory expression induced by a pretest infusion of CNQX into the amygdala and hippocampus or into the entorhinal cortex. Pretest intraentorhinal muscimol also did not influence the effect of pretest intra-amygdala and intrahippocampal CNQX administration. These data indicate that the cells of the entorhinal cortex that are sensitive to pretraining muscimol are not part of the inputs that lead to post-training processing by the amygdala and hippocampus, or to the intervention of the amygdala, hippocampus, and entorhinal cortex in memory expression. The present findings are compatible with the possibility that, instead, the entorhinal cortex may be an output of the amygdala and hippocampus at the time of memory expression.

Memory processing by the limbic system: role of specific neurotransmitter systems.

Experiments using localized infusions into selected brain structures of agonists and antagonists of various synaptic receptors, given before or after behavioral training, have led to the following conclusions: (1) Memory is processed shortly after training in the amygdala, medial septum and hippocampus by glutamatergic NMDA and AMPA receptors activated in that sequence. Cholinergic muscarinic receptors are activated concurrently with the former. GABAA receptors modulated by brain benzodiazepines and by beta-noradrenergic receptors inhibit the process. (2) The sequential involvement of NMDA and AMPA receptors suggests that long-term potentiation (LTP) of the synapses activated by the learning experiences in the hippocampus and/or amygdala and medial septum is the crucial event. Expression of this LTP at the time of testing is necessary for retrieval: AMPA receptor blockade in the hippocampus and amygdala at the time of testing hinders retrieval. This suggests that the LTP underlies the memory process itself. (3) The amygdala, medial septum and hippocampus mediate different types of memory and/or different components of memories. The entorhinal cortex, through mechanisms that require intact NMDA receptors and are inhibited by GABAA receptors, intervenes in post-training memory processing 90-180 min after the other limbic regions. The entorhinal cortex integrates consecutively acquired memories; this role could be maintained by the LTP that is generated after training in the amygdala, hippocampus and medial septum. Post-training intervention of the entorhinal cortex does not occur if this region is inhibited at the time of training.

Memory expression is blocked by the infusion of CNQX into the hippocampus and/or the amygdala up to 20 days after training.

Bilateral infusion of CNQX (0.5 microgram) into the amygdala and the dorsal hippocampus prior to a retention test blocked the expression of step-down inhibitory avoidance in rats 6, 13, or 20 days after training. Retention test performance recovered 90 min after the infusions. Pretest intrahippocampal CNQX (0.5 microgram) blocked the expression of habituation to a novel environment measured 20 days after training. The data suggest that memory expression depends on non-NMDA receptor-mediated mechanisms, perhaps the expression of LTP, up to at least 20 days after acquisition. These mechanisms operate in the hippocampus in both tasks and in the amygdala in the avoidance task.

CNQX infused into rat hippocampus or amygdala disrupts the expression of memory of two different tasks.

The bilateral infusion of CNQX (0.5 or 1.25 micrograms) into the amygdala or dorsal hippocampus 10 min prior to a retention test partially blocked the expression of stepdown inhibitory avoidance in rats 24 h after training. When infused into both the amygdala and the hippocampus at a dose of 0.5 microgram. CNQX caused a complete blockade of the expression of that task. Retention test performance recovered 2 h after the infusions. In rats trained for habituation to a novel environment and tested 24 h later, pretest intrahippocampal CNQX (0.5 microgram) blocked the expression of retention at a dose of 0.5 microgram, and intra-amygdala CNQX (0.5 or 1.25 micrograms) had no effect. The data suggest that, up to at least 1 day after training, memory of the avoidance task depends on glutamate acting on non-NMDA receptors in both the hippocampus and the amygdala, whereas memory of the habituation task depends on non-NMDA receptor activity in the hippocampus but not the amygdala.

Anxiogenic effects of the intraamygdala injection of flumazenil, a benzodiazepine receptor antagonist.

The effects of bilateral intraamygdala microinjection of flumazenil, a benzodiazepine receptor (BZD-R) antagonist, on the exploratory activity in an elevated plus maze were examined in chronically implanted rats. This compound induced a significant decrease in the time spent in the open arms, which is consistent with an anxiogenic action. No effect was observed after intrastriatal injections of flumazenil. Naive rats exposed to the elevated plus maze showed a rapid and selective decline in the content of BZD-like molecules in amygdala (-68%) but not in striatum and hippocampus. These data suggest that the anxiogenic effects of the intraamygdala injection of flumazenil is probably due to the blockade of BZD-like molecules released during the performance.

Amnesia by post-training infusion of glutamate receptor antagonists into the amygdala, hippocampus, and entorhinal cortex.

The blockers of glutamate receptors, aminophosphonovaleric acid (AP5) (5.0 micrograms) and cyano-nitroquinoxaline-dione (CNQX) (0.5 microgram), were infused bilaterally into the amygdala, dorsal hippocampus, or entorhinal cortex of rats through indwelling cannulae 0, 90, 180, or 360 min after step-down inhibitory avoidance training. Animals were tested for retention 24 h after training. In the amygdala or hippocampus, AP5 was amnestic when given 0 min after training and CNQX was amnestic when given 0, 90, or 180 min after training. In the entorhinal cortex, AP5 was amnestic when given 90 or 180 min after training and CNQX had no effect. The results suggest that a phenomenon sensitive first to AP5 and then to CNQX in the amygdala and hippocampus, probably long-term potentiation (LTP), is crucial to post-training memory processing. LTP in these two structures could underlie their role in memory consolidation and could explain the late involvement of the entorhinal cortex in post-training memory processing.