Ricardo C. Da Silva

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.

Intrahippocampal or intraamygdala infusion of KN62, a specific inhibitor of calcium/calmodulin-dependent protein kinase II, causes retrograde amnesia in the rat

We investigated the effect of a bilateral post-training intracerebral infusion of KN62, a specific inhibitor of calcium/calmodulin-dependent protein kinase II (CaM-II), on memory. This enzyme plays a crucial role in the early phases of long-term potentiation. Male Wistar rats were implanted bilaterally with cannulae aimed at the CA1 region of the dorsal hippocampus or at the junction between the central and the basolateral nuclei of the amygdala. After recovery, rats were trained in step-down inhibitory avoidance using a 0.5-mA footshock and tested for retention 24 h later. At various times after training (0, 30, 120, or 240 min for the animals implanted into the hippocampus; 0 or 240 min for the animals implanted in the amygdala) they received, through the cannulae, an infusion of vehicle (0.1% dimethylsulfoxide in water) or KN62 (100 mumol/side). KN62 caused full retrograde amnesia when given 0 min after training into either the amygdala or the hippocampus. When given into the hippocampus 30 min post-training it had a partial amnestic effect. When given 120 min after training into the hippocampus, or 240 min after training into either structure, KN62 had no effect. The data suggest that the early phase of memory requires intact CaM-II activity in the amygdala and hippocampus and support the hypothesis that memory involves long-term potentiation initiated at the time of training in both structures.

Post-training intrahippocampal infusion of protein kinase C inhibitors causes amnesia in rats

This experiment investigated the effect on memory, in rats, of the bilateral intrahippocampal post-training infusion of two different inhibitors of protein kinase C activity, staurosporin and CGP41231. Male Wistar rats were implanted bilaterally with cannulae aimed at the CA1 region of the dorsal hippocampus. After recovery from surgery, they were trained in step-down inhibitory avoidance using a 0.5-mA footshock and tested for retention 24 h later. Immediately or 30, 120, or 180 min after training they received, through the cannulae, infusions of vehicle, staurosporin (1.0 microgram), or CGP41231 (2.5 micrograms). The two drugs caused full retrograde amnesia when given immediately or 30 min post-training, partial amnesia when given 120 min after training, and had no effect when given 180 min after training. The results support the suggestion that memory involves long-term potentiation initiated at the time of training in the hippocampus. Inhibitors of protein kinase C block the development of long-term potentiation when administered in the first 2 h after induction.

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.

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.

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.

Memory expression of habituation and of inhibitory avoidance is blocked by CNQX infused into the entorhinal cortex

Bilateral infusion of CNQX (1.0 micrograms) into the entorhinal cortex 10 min before retention tests blocked the expression of habituation to a novel environment and of step-down inhibitory avoidance in rats. Memory expression was found to be recovered in a second test session carried out 120 min after the infusions in the avoidance task; this was not measured in the habituation task. The data suggest that memory expression of these two tasks depends on non-NMDA receptor-mediated mechanisms, perhaps the expression of LTP, in the entorhinal cortex. Previous experiments had suggested similar mechanisms in the hippocampus and the amygdala. It is possible that, under normal conditions, memory expression may depend on the coordinated activity of the three brain structures.

Effect of antagonists of platelet-activating factor receptors on memory of inhibitory avoidance in rats

Platelet-activating factor (PAF) is present in the brain. It enhances glutamate release and long-term potentiation (LTP) through an action on synaptic membrane receptors sensitive to the antagonist, BN 52021, and has been proposed as a retrograde messenger in the genesis of LTP. In addition, PAF has other, metabolic actions mediated by microsomal receptors sensitive to the antagonist, BN 50730. We investigated the effect on memory of the pre- or post-training infusion of BN 52021 or BN 50730 into the hippocampus and that of BN 52021 in the amygdala and the entorhinal cortex. Male Wistar rats were implanted bilaterally with cannulae aimed at these brain regions. After recovery from surgery, the animals were trained in step-down inhibitory avoidance using a 0.5-mA foot shock and tested for retention 24 h later. BN 52021 (0.5 microgram/side) was amnestic when given into the hippocampus or the amygdala either before or immediately after training but not 30 or 100 min later. BN 52021 was also amnestic when given into the entorhinal cortex 100 but not 0 or 300 min after training. Intrahippocampally administered BN 50730 had no effect on memory. The findings are compatible with the suggestion from previous findings that memory of this task depends on the generation of LTP at the time of training in hippocampus and amygdala and, 90-180 min later, in the entorhinal cortex.

Intrahippocampal, but not intra-amygdala, infusion of an inhibitor of heme oxygenase causes retrograde amnesia in the rat

Zinc protoporhyrin-9 (ZnPP) is an inhibitor of heme oxygenase, the enzyme involved in the biosynthesis of carbon monoxide (CO). CO regulates the activity of glutamatergic synapses and has been proposed to play a role in the early phases of long-term potentiation. The present paper reports on the effect of ZnPP on memory of inhibitory avoidance and of habituation to a novel environment. The bilateral infusion of ZnPP (2 micrograms/side) into the dorsal hippocampus caused amnesia for the inhibitory avoidance task when given before training or 0 or 30 min, but not 60 or 100 min, after training. The immediate post-training intrahippocampal infusion of ZnPP also caused amnesia for the habituation task. The immediate post-training intra-amygdala infusion of ZnPP had no effect on retention of the avoidance task. The data are consistent with the hypotheses that memory involves long-term potentiation initiated at the time of training in the hippocampus, and that hippocampal but not amygdala long-term potentiation may be regulated by CO.

A low power high performance CMOS voltage-mode quaternary full adder

Multiple-valued logic, despite of all its theoretical potentialities, has not provided real advantages for arithmetic circuits when compared to the binary equivalent ones until now. This paper shows a new efficient method to implement quaternary logic arithmetic circuits using multi-threshold transistors, where 3 power supply lines are used to perform quaternary circuits with low power consumption and high performance. As a demonstration, a quaternary full adder is described in TSMC 0.18mum technology and compared to regular binary circuits, presenting a 76% reduction in power consumption, and an improvement of 15% regarding speed with a 20% area overhead