Differential neurophysiological correlates of retrieval of consolidated and reconsolidated memories in humans: An ERP and pupillometry study.

Consolidated memories can return to a labile state if they are reactivated by unpredictable reminders. To persist, active memories must be re-stabilized through a process known as reconsolidation. Although there is consistent behavioral evidence about this process in humans, the retrieval process of reconsolidated memories remains poorly understood. In this context, one fundamental question is whether the same or different neurophysiological mechanisms are involved in retrieval of consolidated and reconsolidated memories. Because it has been demonstrated that the exposure to the reconsolidation process may restructure and strengthen memories, we hypothesized distinct neurophysiological patterns during retrieval of reconsolidated memories. In addition, we hypothesized that interfering with the reconsolidation process using a new learning can prevent these neurophysiological changes. To test it, consolidated, reconsolidated and declarative memories whose reconsolidation process was interfered (i.e., picture-word pairs) were evaluated in humans in an old/new associative recall task while the brain activity and the pupillary response were recorded using electroencephalography and eyetracking. Our results showed that retrieval of reconsolidated memories elicits specific patterns of brain activation, characterized by an earlier peak latency and a smaller magnitude of the left parietal ERP old/new effect compared to memories that were only consolidated or whose reconsolidation process was interfered by a new learning. Moreover, our results demonstrated that only retrieval of reconsolidated memories is associated with a late reversed mid-frontal effect in a 600-690 time window. Complementarily, memories that were reactivated showed an earlier peak latency of the pupil old/new effect compared to non-reactivated memories. These findings support the idea that reconsolidation has an important impact in how memories are retrieved in the future, showing that retrieval of reconsolidated memories is partially supported by specific brain mechanisms.

Enhancing a declarative memory in humans: the effect of clonazepam on reconsolidation.

A consolidated memory recalled by a specific reminder can become unstable (labile) and susceptible to facilitation or impairment for a discrete period of time. This labilization phase is followed by a process of stabilization called reconsolidation. The phenomenon has been shown in diverse types of memory, and different pharmacological agents have been used to disclose its presence. Several studies have revealed the relevance of the GABAergic system to this process. Consequently, our hypothesis is that the system is involved in the reconsolidation of declarative memory in humans. Thus, using our verbal learning task, we analyzed the effect of benzodiazepines on the re-stabilization of the declarative memory. On Day 1, volunteers learned an association between five cue- response-syllables. On Day 2, the verbal memory was labilized by a reminder presentation, and then a placebo capsule or 0.25 mg or 0.03 mg of clonazepam was administered to the subjects. The verbal memory was evaluated on Day 3. The volunteers who had received the 0.25 mg clonazepam along with the specific reminder on Day 2, exhibited memory improvement. In contrast, there was no effect when the drug was given without retrieval, when the memory was simply retrieved instead of being reactivated or when short-term memory testing was performed 4 h after reactivation. We discuss the GABAergic role in reconsolidation, which shows a collateral effect on other memories when the treatment is aimed at treating anxiety disorders. Further studies might elucidate the role of GABA in the reconsolidation process associated with dissimilar scenarios. This article is part of a Special Issue entitled 'Cognitive Enhancers'.

Memory consolidation and reconsolidation in an invertebrate model: the role of the GABAergic system.

Consolidation theory assumes that memories are labile during a limited time window after acquisition, but as time passes, memories become stable and resistant to amnesic agents. However, the vision of immutable memories after consolidation has been challenged. Thus, after the presentation of a reminder, the reactivated old memories become labile and again susceptible to amnesic treatments. This process implies a re-stabilization phase, usually referred to as reconsolidation. Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter both in the Central nervous system (CNS) and in the periphery. A considerable amount of evidence has arisen from different studies regarding the role of the GABA(A) receptor in diverse behavioral paradigms and tasks. Here, we investigate the role of the GABAergic system on both memory consolidation and reconsolidation phases by using the memory paradigm of the crab Chasmagnathus. In order to achieve such a goal, we design pharmacological-behavioral experiments, which include the administration of classic agonist (muscimol) and antagonist (bicuculline) of the mammals GABA(A) receptors. The current results show that the systemic administration of muscimol impairs the consolidation and reconsolidation processes. In contrast, the administration of bicuculline improves the consolidation and reconsolidation processes. Furthermore, the co-administration of both drugs blocks the agonist amnesic effect on the consolidation phase. The ubiquity of the neurotransmitter and its receptors in the animal taxa allows us to use the classic agonist-and-antagonist administration procedure in this invertebrate. Thus, all the results reported in this paper can be judged as a result of the modulation exerted by the functional state of the GABAergic system in the CNS. To conclude, the results obtained in this report with an invertebrate model represent additional evidences supporting the view that some molecular mechanisms subserving different memory phases could be the basic tools employed by phylogenetically disparate animals.

Human reconsolidation does not always occur when a memory is retrieved: the relevance of the reminder structure.

Memory reconsolidation is defined as a process in which the retrieval of a previously consolidated memory returns to a labile state which is then subject to stabilization. The reminder is the event that begins with the presentation of the learned cue and triggers the labilization-reconsolidation process. Since the early formulation of the hypothesis, several controversial items have arisen concerning the conditions that define reconsolidation. It is herein proposed that two diagnostic features characterize reconsolidation, namely: the labilization of the reactivated memory and the specificity of the reminder structure. To study this proposal, subjects received two different training sessions on verbal material on Day 1 and Day 2, respectively. Finally, they were tested for the first and second acquired memories on Day 3. It is demonstrated that the human declarative memory fulfills the two requirements that define the process. First, the reactivated memory is impaired by a new learning only when it is given closely after the reminder, revealing that the memory is labilized. Second, the omission of at least one of the reminder's components prevents labilization. Therefore, results show that the new learning fails to produce an amnesic effect on the target memory either when the reminder omits the learned cue or includes the beginning of the reinforcement.

Context shift and protein synthesis inhibition disrupt long-term habituation after spaced, but not massed, training in the crab Chasmagnathus.

An opaque screen moving overhead elicits an escape response in the crab Chasmagnathus that after a few presentations habituates for a long period (long-term habituation, LTH). Previous results suggested that spaced (15 trials separated by 171 s) and massed training (300 trials without rest interval) were correlated with two different memory components of LTH. The present experiments were aimed at further studying the mechanisms subserving these components. Results indicate that LTH acquired by spaced but not by massed training is blocked either by a training-to-testing context shift or by cycloheximide (15-25 microg) pre- or posttraining injection and that LTH after spaced training persists for longer time (5 days) than after massed training (2 days). A model based on these results that distinguishes two LTH-memory components is proposed: a (context-signal) LTH yielded by spaced training, dependent of context, sensitive to cycloheximide (CYX), and long lasting; and a (signal) LTH yielded by massed training, dependent only on the signal invariance, insensitive to CYX, and shorter lasting.

Effects of activation and inhibition of cAMP-dependent protein kinase on long-term habituation in the crab Chasmagnathus.

On sudden presentation of a danger stimulus, the crab Chasmagnathus elicits an escape response that habituates promptly and for a long period. We have previously reported that administration of a cAMP-permeable analog (CPT-cAMP) along with a phosphodiesterase inhibitor (IBMX) improves long-term habituation (LTH). In present experiments we studied the effect of systemic administration of the protein kinase A (PKA) activator Sp-5,6-DCl-cBIMPS and that of the PKA inhibitor Rp-8-Cl-cAMPS on LTH tested 24 h after a weak training protocol (5 trials of danger stimulus presentation) or a strong training protocol (15-30 trials), respectively. A 50 microliters pre-training injection of 75 microM Sp-5,6-DCl-cBIMPS, and to a lesser degree of 25 microM, improved retention of the habituated response but not affect short-term habituation (STH). Like pre-training injection, post-training administration of Sp-5,6-DCl-cBIMPS proved to exert a facilitatory action on retention though with 75 microM dose only. Conversely, both pre- and post-training injection of 25 microM Rp-8-Cl-cAMPS impaired LTH without affecting STH. Thus, the PKA activator Sp-5,6-DCl-cBIMPS enables a weak training to produce LTH while the PKA inhibitor Rp-8-Cl-cAMPS impairs LTH when a strong training is given. Activation of crab PKA by Sp-5,6-DCl-cBIMPS and its inhibition by Rp-8-Cl-cAMPS were assessed using an in vitro PKA activity assay. These results provide independent evidences supporting the view that PKA plays a key role in long-term memory storage in this learning paradigm.

Inhibitors of protein and RNA synthesis block context memory and long-term habituation in the crab Chasmagnathus.

The crab Chasmagnathus granulatus reacts to a shadow passing overhead (a danger stimulus) with an escape response that habituates quickly and for at least 5 days. Recently, it has been reported that cycloheximide (CY) disrupts this long-term habituation and the corresponding context memory. In the present article, experiments with CY and an inhibitor of RNA synthesis, actinomycin-D (ACT), were parallelly conducted. An injection of CY (20 micrograms) or ACT (0.62 microgram) reduced the incorporation of [14C]-aminoacid into cerebral plus thoracic ganglia by 80% for 2 h and 59.7% for 1 h, respectively, but no inhibition was found at 24 h. Both ACT (0.62 microgram) and CY (20 micrograms) administered immediately after training (15 trials with the danger stimulus) impaired either long-term habituation or context memory when tested at 24 h. Because ACT and CY have in common only their direct or indirect inhibitory effect on protein synthesis, this finding is considered as an additional evidence that long-term memory in Chasmagnathus requires de novo protein synthesis. However, pretraining ACT or CY impaired context memory at 24 h but not long-term habituation. Such a disparity is explained by an unspecific attenuating effect upon the response, attributed to drug x training interaction. Neither ACT nor CY affected short-term habituation.

Acute administration of a permeant analog of cAMP and a phosphodiesterase inhibitor improve long-term habituation in the crab Chasmagnathus.

A shadow passing overhead acts as a danger stimulus and elicits an escape response in the crab Chasmagnathus that habituates promptly and for a long period. Robust retention is shown at 24 h after 15 trials of shadow presentation or at 120 h after 30 trials, but no significant retention is disclosed at 24 h after 5 trials or at 72 h after 15. A cocktail of the cAMP membrane permeable analog 8-(4-chlorophenylthio)-cAMP (CPT-cAMP), plus the phosphodiesterase inhibitor isobutyl methylxanthine (IBMX), was given by systemic administration. Pretraining injection of the cocktail (25 or 50 microM, 15 min before a 5-trial session) failed to affect short-term habituation, but induced significant retention when tested at 24 h. This facilitatory effect was not shown when a lower dose (5 microM) was used. A post-training injection of 25 microM, immediately after a 5- or 15-trial session, induced retention when tested at 24 or 72 h, respectively. Thus, the administration of CPT-cAMP + IBMX during acquisition of a habituated response or immediately after, improves long-term habituation, a result supporting the view that an increase in the cAMP level is one of the steps in long-term memory consolidation.

Angiotensin II enhances long-term memory in the crab Chasmagnathus.

An opaque screen moving overhead provokes an escape response in the crab Chasmagnathus granulatus that habituates after a few presentations of the eliciting stimulus. Fifteen trials with a 180-s intertrial interval or 30 trials with a 90-s interval (strong training protocol) ensures long-term habituation (LTH) of the response for 24 h, whereas 10 trials (weak training protocol) fail to induce it. However, robust LTH is obtained when crabs are injected with human angiotensin (All; 50 pmol) immediately after a weak training protocol. This memory-enhancing effect of All is dose-dependent, reversible by saralasin (5 pmol), and vanishes either when the weak training protocol is reduced to only five trials, or when the peptide is given before training or 1 h after. LTH is impaired by saralasin (5 pmol) administered before or after the strong training protocol, but no amnestic effect is disclosed when the antagonist is given 1 h after. On the other hand, both All-like immunoreactivity and angiotensin-converting enzyme-like activity are described in diverse tissues of Chasmagnathus, namely, in gills and in both thoracic and supraesophageal ganglia. Results support the view that some components of the renin-angiotensin system and their influence on memory might have emerged early in evolution.

Cycloheximide inhibits context memory and long-term habituation in the crab Chasmagnathus.

A shadow moving over head elicits an escape response in the crab Chasmagnathus that habituates promptly and for a long period. The effect of the protein synthesis inhibitor cycloheximide (CY) on this long-term memory was analyzed. Two hours after injection, 10 micrograms CY inhibited [14C]-amino amino acid incorporation into cerebral plus thoracic ganglia by 88% and 20 micrograms by 92%, but no inhibition was found at 24 h. A single injection of 10-20 micrograms CY given 30 min before training, failed to affect the short-term habituation. Similar doses impaired both context memory (CM) and long-term habituation (LTH) when tested at 72 and 120 h but only CM at 24 h. Such a disparity was explained by an unspecific depressing effect upon the response, attributed to an interaction between CY and training. The hypothesis was confirmed, because CY injected immediately after training disclosed amnestic effect at 24 h on both CM and LTH. A similar effect was proven when animals were injected at 2 h but not at 6 h after training. Results from experiments with pretraining and pretesting injections put aside a state-dependence or retrieval deficit effects of the drug. Taken together, findings of this article argue strongly for de novo protein synthesis as a mechanism of LTH and for the close relation between CM and LTH.

Acute administration of angiotensin II improves long-term habituation in the crab Chasmagnathus.

A shadow moving overhead acts as a danger stimulus and elicits an escape response in the crab Chasmagnathus granulatus that habituates after 15 trials and for a long period. A shorter training of ten trials fails to induce long-term habituation; however, a good retention of the habituated response is manifest after a 24-h interval when angiotensin II (AII) (10(-6) M, 3 ng/g) is injected post-training. By contrast, no amnestic effect of AII was found even though high doses were administered. The facilitatory effect of AII is suppressed by saralasin (10(-7) M, 0.3 ng/g), a specific angiotensin II receptor antagonist. Results are considered as suggesting that angiotensin on memory processes might have emerged early in evolution.