Fernando Benetti

Behavioral tagging of extinction learning

Extinction of contextual fear in rats is enhanced by exposure to a novel environment at 1–2 h before or 1 h after extinction training. This effect is antagonized by administration of protein synthesis inhibitors anisomycin and rapamycin into the hippocampus, but not into the amygdala, immediately after either novelty or extinction training, as well as by the gene expression blocker 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole administered after novelty training, but not after extinction training. Thus, this effect can be attributed to a mechanism similar to synaptic tagging, through which long-term potentiation can be enhanced by other long-term potentiations or by exposure to a novel environment in a protein synthesis-dependent fashion. Extinction learning produces a tag at the appropriate synapses, whereas novelty learning causes the synthesis of plasticity-related proteins that are captured by the tag, strengthening the synapses that generated this tag.

Effects of acute and chronic physical exercise and stress on different types of memory in rats

Here we study the effect of acute and chronic physical exercise in a treadmill and of daily stress (because forced exercise involves a degree of stress) during 2 or 8 weeks on different types of memory in male Wistar rats. The memory tests employed were: habituation in an open field, object recognition and spatial learning in the Morris water maze. Daily foot-shock stress enhanced habituation learning after 2 but not after 8 weeks; it hindered both short- (STM) and long-term memory (LTM) of the recognition task at 2 weeks but only STM after 8 weeks and had no effect on spatial learning after either 2 or 8 weeks. Acute but not chronic exercise also enhanced habituation in the open field and hindered STM and LTM in the recognition task. Chronic exercise enhanced one important measure of spatial learning (latency to escape) but not others. Our findings indicate that some care must be taken when interpreting effects of forced exercise on brain parameters since at least part of them may be due to the stress inherent to the training procedure.

Physical exercise can reverse the deficit in fear memory induced by maternal deprivation

Maternal deprivation during the first 10 days of life induces significant behavioral alterations in rodents which persist through adulthood. Physical exercise reduces the cognitive deficits associated with pharmacologic and pathological conditions. Here we investigated whether forced physical exercise alters memory deficits caused by postnatal maternal deprivation. Male rats were divided into four groups: (1) control, (2) deprived, (3) exercised, and (4) deprived+exercised. In groups 2 and 4, pups were deprived from their mothers for 3h/day during the first 10 days post-birth. In groups 3 and 4, from postnatal day 45 (PND-45) on, animals were submitted to forced treadmill exercise. At adulthood, animals were submitted to four different behavioral tasks: open field, Morris water maze (MWM), object recognition (OR) and inhibitory avoidance (IA). Maternal deprivation had no effect on open field behavior, but disrupted memory in the three other tasks. Physical exercise alone had no effect, except for a slight enhancement of MWM learning. Importantly, physical exercise reversed the deficit of IA and reduced the deficit of spatial memory but not that of OR seen in deprived animals. It is possible that physical exercise may counteract the influence of maternal deprivation on neurohumoral or hormonal memory modulatory systems related to stress. Indeed, the decreasing order of the effect of exercise on the memory disturbances induced by deprivation roughly follows the descending degree of stress associated with each task (IA>MWM>OR). Maternal deprivation is known to hinder hormonal mechanisms involved in coping with stress.

Histaminergic mechanisms for modulation of memory systems.

Encoding for several memory types requires neural changes and the activity of distinct regions across the brain. These areas receive broad projections originating in nuclei located in the brainstem which are capable of modulating the activity of a particular area. The histaminergic system is one of the major modulatory systems, and it regulates basic homeostatic and higher functions including arousal, circadian, and feeding rhythms, and cognition. There is now evidence that histamine can modulate learning in different types of behavioral tasks, but the exact course of modulation and its mechanisms are controversial. In the present paper we review the involvement of the histaminergic system and the effects histaminergic receptor agonists/antagonists have on the performance of tasks associated with the main memory types as well as evidence provided by studies with knockout models. Thus, we aim to summarize the possible effects histamine has on modulation of circuits involved in memory formation.

Hippocampal molecular mechanisms involved in the enhancement of fear extinction caused by exposure to novelty.

Significance Within a restricted time window, a brief exposure to a novel environment enhances the extinction of contextual fear. This can be explained by a hippocampal process of behaviorally induced synaptic tagging and capture. Here, we report that the effect requires glutamate NMDA receptors and L-voltage–dependent calcium channels and involves the activation of calcium/calmodulin-dependent protein kinase II, in addition to both ribosomal and nonribosomal protein synthesis. All these mechanisms operate only when the proteasomal-ubiquitin protein degradation system is intact, which suggests that they depend on synaptic protein turnover. Extinction enhancement by novelty is of great potential importance in the treatment of fear memories, such as those of posttraumatic stress disorder; the treatments of choice for such conditions are based on extinction procedures. Exposure to a novel environment enhances the extinction of contextual fear. This has been explained by tagging of the hippocampal synapses used in extinction, followed by capture of proteins from the synapses that process novelty. The effect is blocked by the inhibition of hippocampal protein synthesis following the novelty or the extinction. Here, we show that it can also be blocked by the postextinction or postnovelty intrahippocampal infusion of the NMDA receptor antagonist 2-amino-5-phosphono pentanoic acid; the inhibitor of calcium/calmodulin-dependent protein kinase II (CaMKII), autocamtide-2–related inhibitory peptide; or the blocker of L-voltage–dependent calcium channels (L-VDCCs), nifedipine. Inhibition of proteasomal protein degradation by β-lactacystin has no effect of its own on extinction or on the influence of novelty thereon but blocks the inhibitory effects of all the other substances except that of rapamycin on extinction, suggesting that their action depends on concomitant synaptic protein turnover. Thus, the tagging-and-capture mechanism through which novelty enhances fear extinction involves more molecular processes than hitherto thought: NMDA receptors, L-VDCCs, CaMKII, and synaptic protein turnover.

The evidence for hippocampal long-term potentiation as a basis of memory for simple tasks

Long-term potentiation (LTP) is the enhancement of postsynaptic responses for hours, days or weeks following the brief repetitive afferent stimulation of presynaptic afferents. It has been proposed many times over the last 30 years to be the basis of long-term memory. Several recent findings finally supported this hypothesis: a) memory formation of one-trial avoidance learning depends on a series of molecular steps in the CA1 region of the hippocampus almost identical to those of LTP in the same region; b)hippocampal LTP in this region accompanies memory formation of that task and of another similar task. However, CA1 LTP and the accompanying memory processes can be dissociated, and in addition plastic events in several other brain regions(amygdala, entorhinal cortex, parietal cortex) are also necessary for memory formation of the one-trial task, and perhaps of many others.

Memory reconsolidation and its maintenance depend on L-voltage-dependent calcium channels and CaMKII functions regulating protein turnover in the hippocampus.

Immediate postretrieval bilateral blockade of long-acting voltage–dependent calcium channels (L-VDCCs), but not of glutamatergic NMDA receptors, in the dorsal CA1 region of the hippocampus hinders retention of long-term spatial memory in the Morris water maze. Immediate postretrieval bilateral inhibition of calcium/calmodulin-dependent protein kinase (CaMK) II in dorsal CA1 does not affect retention of this task 24 h later but does hinder it 5 d later. These two distinct amnesic effects are abolished if protein degradation by proteasomes is inhibited concomitantly. These results indicate that spatial memory reconsolidation depends on the functionality of L-VDCC in dorsal CA1, that maintenance of subsequent reconsolidated memory trace depends on CaMKII, and these results also suggest that the role played by both L-VDCC and CaMKII is to promote the retrieval-dependent, synaptically localized enhancement of protein synthesis necessary to counteract a retrieval-dependent, synaptic-localized enhancement of protein degradation, which has been described as underlying the characteristic labilization of the memory trace triggered by retrieval. Thus, conceivably, L-VDCC and CaMKII would enhance activity-dependent localized protein renewal, which may account for the improvement of the long-term efficiency of the synapses responsible for the maintenance of reactivated long-term spatial memory.

Astrocytes as a target for neuroprotection: Modulation by progesterone and dehydroepiandrosterone.

Stroke and traumatic injuries of the brain and spinal cord are major public health issues. In the last few decades, hundreds of clinical trials with patients suffering from these conditions have been done, however, most of them had not succeeded and there is still the need to develop more effective treatments for these conditions. Astrocytes play critical roles in the development, function and survival of neurons in the central nervous system. These cells are implicated in the pathophysiology and in the response to several neuropathological conditions and may represent potential cell targets for neuroprotective strategies. Progesterone and dehydroepiandrosterone (DHEA) are neuroactive steroids that modulate neuronal and astroglial function and have neuroprotective effects in different experimental models, being potential candidates to the development of new therapeutic approaches for brain and spinal cord injuries. The aim of this review is to discuss the role of astrocytes in the pathophysiology of brain and spinal cord injuries and how they could be modulated by progesterone and DHEA for the treatment of these conditions.

Histamine infused into basolateral amygdala enhances memory consolidation of inhibitory avoidance.

The role of the basolateral amygdala (BLA) in the consolidation of aversive memory is well established. Here we investigate the involvement of the histaminergic system in BLA on this variable. Rats were chronically implanted with bilateral cannulae in the BLA and after recovery were trained in a one-trial step-down inhibitory avoidance task. Immediately after training histaminergic compounds either alone or in combination were infused through the cannulae. Memory was assessed in test sessions carried out 24 h after the training session. Post-training histamine (1-10 nmol; 0.5 μl/side) enhanced consolidation and the histamine H₃ receptor antagonist thioperamide (50 nmol; 0.5 μl/side) impaired memory consolidation. The effect was shared by the histamine N-methyltransferase inhibitor SKF-91844 (50 nmol; 0.5 μl/side) as well as by the H₃ receptor agonist imetit (10 nmol; 0.5 μl/side). The promnesic action of histamine was unaffected by the H₁ receptor antagonist pyrilamine (50 nmol; 0.5 μl/side). The H1 receptor agonist pyridylethylamine (10 nmol; 0.5 μl/side), the H₂ agonist dimaprit (10 nmol; 0.5 μl/side) and the H₂ antagonist ranitidine (50 nmol; 0.5 μl/side) were ineffective. Histaminergic compounds infused into the BLA had no effect on open-field or elevated plus-maze behaviour. The data show that histamine induces a dose-dependent mnemonic effect in rats and indicate that this reflects a role of endogenous histamine in the BLA mediated by H₃ receptors.

Histamine in the basolateral amygdala promotes inhibitory avoidance learning independently of hippocampus

Significance Integrity of the brain histaminergic system is necessary for long-term memory (LTM) but not short-term memory of step-down inhibitory avoidance (IA). Histamine depletion in hippocampus or basolateral amygdala (BLA) impairs LTM of that task. Histamine infusion into either structure restores LTM in histamine-depleted rats. The restoring effect in BLA occurs even when hippocampal activity was impaired. Cyclic adenosine monophosphate (cAMP) responsive-element-binding protein phosphorylation correlates anatomically and temporally with histamine-induced memory recall. Thus, histaminergic neurotransmission appears critical to provide the brain with the plasticity necessary for IA through recruitment of alternative circuits. Our findings indicate that the histaminergic system comprises parallel, coordinated pathways that provide compensatory plasticity when one brain structure is compromised. Recent discoveries demonstrated that recruitment of alternative brain circuits permits compensation of memory impairments following damage to brain regions specialized in integrating and/or storing specific memories, including both dorsal hippocampus and basolateral amygdala (BLA). Here, we first report that the integrity of the brain histaminergic system is necessary for long-term, but not for short-term memory of step-down inhibitory avoidance (IA). Second, we found that phosphorylation of cyclic adenosine monophosphate (cAMP) responsive-element-binding protein, a crucial mediator in long-term memory formation, correlated anatomically and temporally with histamine-induced memory retrieval, showing the active involvement of histamine function in CA1 and BLA in different phases of memory consolidation. Third, we found that exogenous application of histamine in either hippocampal CA1 or BLA of brain histamine-depleted rats, hence amnesic, restored long-term memory; however, the time frame of memory rescue was different for the two brain structures, short lived (immediately posttraining) for BLA, long lasting (up to 6 h) for the CA1. Moreover, long-term memory was formed immediately after training restoring of histamine transmission only in the BLA. These findings reveal the essential role of histaminergic neurotransmission to provide the brain with the plasticity necessary to ensure memorization of emotionally salient events, through recruitment of alternative circuits. Hence, our findings indicate that the histaminergic system comprises parallel, coordinated pathways that provide compensatory plasticity when one brain structure is compromised.