Spatial learning in rats is impaired by microinfusions of protein kinase C-gamma antisense oligodeoxynucleotide within the nucleus accumbens

The nucleus accumbens (NAcc) has been shown to play a role in motor and spatial learning. Protein kinase C (PKC) has been implicated in the mechanisms of initiation and maintenance of long-term potentiation that is thought to be involved in the storage of long-term memory. In the present study, the importance of de novo synthesis of PKC-gamma within the NAcc in the acquisition and retention of spatial discrimination learning was assessed using an antisense knockdown approach. Separate groups of Long-Evans rats were exposed to acute microinfusions (6microg/microl) of PKC-gamma antisense oligodeoxynucleotide (AS-ODN), control oligodeoxynucleotide (C-ODN) or vehicle into the NAcc at 24 and 3h before each training session. Behavioral findings showed that the blockade of NAcc-PKC-gamma translation caused impairments in the early phase of learning and retention of spatial information. Biochemical experiments showed that PKC-gamma expression was reduced and Ca(2+)/phospholipid-dependent protein kinase C (PKC) activity was blocked significantly in the AS-ODN-treated rats in comparison with control rats. The present findings suggest that NAcc-PKC-gamma plays a role during the early acquisition of spatial learning. Also, retention test results suggest that NAcc-PKC-gamma may be working as an intermediate factor involved in the onset of molecular mechanisms necessary for spatial memory consolidation within the NAcc.     

Individual differences in dopamine efflux in nucleus accumbens shell and core during instrumental learning

Combined activation of dopamine D1- and NMDA-glutamate receptors in the nucleus accumbens has been strongly implicated in instrumental learning, the process in which an individual learns that a specific action has a wanted outcome. To assess dopaminergic activity, we presented rats with two sessions (30 trials each) of a one-lever appetitive instrumental task and simultaneously measured dopamine efflux in the shell and core accumbens subareas using in vivo microdialysis. Dopamine efflux was increased during each session in all areas. The behavioral performance of the rats in the second session led us to divide them into a learning group (>90% correct trials) and a non-learning group. In the first session, the rats of the learning group showed significantly higher increases. The difference was most pronounced in the shell. In the second session, the dopamine increase was similar in both groups, although the learning groups now pressed the lever about three times more often and consequently obtained more rewards. We conclude that task-related activation of dopamine efflux is different between learning and non-learning rats only during the learning phase. These results support the pharmacological evidence that dopamine is of particular importance during the instrumental learning process.     

Open field habituation learning is improved by nicotine and attenuated by mecamylamine administered posttrial into the nucleus accumbens

Using the paradigm of habituation learning in the open field, we tested the effects of unilateral microinjections of the agonist nicotine (8.0, 40.0, and 80.0 microg) and the nicotine receptor antagonist mecamylamine (0.1, 1.0, 10.0 microg) into the core of the nucleus accumbens. When injected posttrial, that is, immediately after the first exposure to the open field, nicotine dose-dependently enhanced behavioral habituation during the test on the following day, indicating a facilitation of memory, whereas mecamylamine impaired habituation at the highest dose, but not at the two lower doses. When injected 5 h after the learning trial, nicotine (40 microg) and mecamylamine (10 microg) impaired habituation on the subsequent day. A control experiment did not provide evidence for possible proactive effects of mecamylamine. These findings are discussed with respect to the possible behavioral functions of cholinergic, and especially nicotinic, mechanisms in the nucleus accumbens. They may also be relevant for understanding cholinergic-linked psychopathologies such as Alzheimer's disease, since the nucleus accumbens is one of the sites where cholinergic neurons are lost in this neurodegenerative disease.     

The role of nucleus accumbens dopamine in outcome encoding in instrumental and Pavlovian conditioning

Considerable evidence suggests that dopamine in the core subregion of the nucleus accumbens is not only involved in Pavlovian conditioning but also supports instrumental performance. However, it is largely unknown whether NAc dopamine is required for outcome encoding which plays an important role both in Pavlovian stimulus-outcome learning and instrumental action-outcome learning. Therefore, we tested rats with 6-hydroxydopamine (6-OHDA) induced dopamine depletion of the NAc core for their sensitivity to outcome devaluation in a Pavlovian and an instrumental task. Results indicate that 6-OHDA-lesioned animals were sensitive to outcome devaluation in an instrumental task. This finding provides support to the notion that NAc core dopamine may not be crucial in encoding action-outcome associations. However, during instrumental conditioning lever pressing rates in 6-OHDA-lesioned animals were markedly lower which could reflect an impaired behavioral activation. By contrast, after outcome-specific devaluation in a Pavlovian task, performance in 6-OHDA-lesioned animals was impaired, i.e. their magazine-directed responding was non-selectively reduced. One possibility to explain non-selective responding is that NAc core DA depletion impaired the ability of conditioned stimuli to activate the memory of the current value of the reinforcer.     

Dorsal and ventral striatal protein synthesis inhibition affect reinforcer valuation but not the consolidation of instrumental learning

The evidence for a role of the striatum in the acquisition of uncued instrumental responding is ambiguous. It has been shown that post-session infusions of anisomycin into the core of the nucleus accumbens (NAcc) impaired instrumental acquisition, but pre-training lesions of the NAcc suggest that it is not necessary. Recently, we demonstrated that the infusion of anisomycin into the anterior cingulate cortex impaired instrumental acquisition indirectly through a taste aversion. Thus, we hypothesized that post-session anisomycin infusions into the NAcc affected instrumental acquisition through an effect on reinforcer valuation. For the dorsal striatum, both post-session infusions of anisomycin and pre-training lesion studies suggest that neither the dorsolateral nor the dorsomedial striatum is necessary for the acquisition of instrumental responding. However, it has not been attempted to block plasticity in both regions concurrently, and we hypothesized that both regions independently contribute to acquisition through goal-directed and habitual learning. In the current experiments, we first replicated the effect of unprotected post-session anisomycin infusions into the NAcc on instrumental acquisition. Subsequently, we investigated the effect of protein synthesis inhibition in the NAcc and dorsomedial and dorsolateral striatum concurrently on instrumental acquisition, critically controlling for effects on reinforcer valuation. The anisomycin infusions induced an aversive state, but did not affect instrumental acquisition.     

Observational learning in mice can be prevented by medial prefrontal cortex stimulation and enhanced by nucleus accumbens stimulation

The neural structures involved in ongoing appetitive and/or observational learning behaviors remain largely unknown. Operant conditioning and observational learning were evoked and recorded in a modified Skinner box provided with an on-line video recording system. Mice improved their acquisition of a simple operant conditioning task by observational learning. Electrical stimulation of the observer's medial prefrontal cortex (mPFC) at a key moment of the demonstration (when the demonstrator presses a lever in order to obtain a reward) cancels out the benefits of observation. In contrast, electrical stimulation of the observer's nucleus accumbens (NAc) enhances observational learning. Ongoing cognitive processes in the demonstrator could also be driven by electrical stimulation of these two structures, preventing the proper execution of the ongoing instrumental task (mPFC) or stopping pellet intake (NAc). Long-term potentiation (LTP) evoked in these two cortical structures did not prevent the acquisition or retrieval process--namely, mPFC and/or NAc stimulation only prevented, or modified, the ongoing behavioral process. The dorsal hippocampus was not involved in either of these two behavioral processes. Thus, both ongoing observational learning and performance of an instrumental task require the active contribution of the mPFC and/or the NAc.     

Speech segmentation by statistical learning is supported by domain-general processes within working memory

The purpose of this study was to examine the extent to which working memory resources are recruited during statistical learning (SL). Participants were asked to identify novel words in an artificial speech stream where the transitional probabilities between syllables provided the only segmentation cue. Experiments 1 and 2 demonstrated that segmentation performance improved when the speech rate was slowed down, suggesting that SL is supported by some form of active processing or maintenance mechanism that operates more effectively under slower presentation rates. In Experiment 3 we investigated the nature of this mechanism by asking participants to perform a two-back task while listening to the speech stream. Half of the participants performed a two-back rhyme task designed to engage phonological processing, whereas the other half performed a comparable two-back task on un-nameable visual shapes. It was hypothesized that if SL is dependent only upon domain-specific processes (i.e., phonological rehearsal), the rhyme task should impair speech segmentation performance more than the shape task. However, the two loads were equally disruptive to learning, as they both eradicated the benefit provided by the slow rate. These results suggest that SL is supported by working-memory processes that rely on domain-general resources.     

Outcome-specific transfer between predictive and instrumental learning is unaffected by extinction but reversed by counterconditioning in human participants

Three experiments were conducted to explore the effects of different interference treatments upon outcome-specific transfer from predictive learning to instrumental responding. A computer game was designed in which participants had to defend Andalusia from navy and air-force attacks. Participants learned the relationship between two instrumental responses (two keys in a standard keyboard) and two different outcomes (destruction of the ships or destruction of the planes). Then, they learned to predict which of two different cues predicted either outcome. Finally, participants were allowed to give either of the two instrumental responses in the presence of each cue. Outcome-specific transfer was shown as a preference for the response that shared the outcome with the current cue. Extinction of the cue before the transfer test had no effect upon transfer, regardless of the level of extinction (Experiments 1-3). However, pairing the cue with the alternative outcome (counterconditioning) reversed the outcome-based transfer effect (Experiment 3). The implications of these results for the contents of extinction in human predictive learning are discussed.     

Differential activity profile of cAMP-dependent protein kinase isoforms during long-term memory consolidation in the crab Chasmagnathus

The isoforms of cAMP-dependent protein kinase (PKA) show distinct biochemical properties and subcellular localization, suggesting different physiological functions, and conferring the fine-tuning between the activation of cAMP-PKA cascade and the cellular response. The critical role of PKA in memory and synaptic plasticity has been extensively demonstrated both in vertebrates and invertebrates, but the role of PKA isoforms is a matter of debate. Here we present experimental data showing differential PKA activation profiles after two different experiences: an instance of associative contextual learning (context-signal learning) and a single exposure to a novel context, both in the learning and memory model of the crab Chasmagnathus. Differences were found in the temporal course of activation and in the involvement of PKA isoforms. We found increased PKA activity immediately and 6 h after context-signal training correlating with the critical periods during which pharmacological inhibition of PKA disrupts memory formation. In contrast, PKA activity increased immediately but not 6 h after single exposure to a novel context. The amounts of PKA I and PKA II holoenzymes were analyzed to determine changes in holoenzyme levels and/or differential activation induced by both experiences. Results indicate that context-induced PKA activation is at least in part due to PKA II, and that PKA activation 6 h after context-signal learning coincides with an increase in the total level of PKA I. Considering the higher sensitivity of PKA I to cAMP, its increment can account for the PKA activation found 6 h after training and is proposed as a novel mechanism providing the prolonged PKA activation during memory consolidation.     

Associative learning performance is impaired in zebrafish (Danio rerio) by the NMDA-R antagonist MK-801

The zebrafish is gaining popularity in behavioral neuroscience perhaps because of a promise of efficient large scale mutagenesis and drug screens that could identify a substantial number of yet undiscovered molecular players involved in complex traits. Learning and memory are complex functions of the brain and the analysis of their mechanisms may benefit from such large scale zebrafish screens. One bottleneck in this research is the paucity of appropriate behavioral screening paradigms, which may be due to the relatively uncharacterized nature of the behavior of this species. Here we show that zebrafish exhibit good learning performance in a task adapted from the mammalian literature, a plus maze in which zebrafish are required to associate a neutral visual stimulus with the presence of conspecifics, the rewarding unconditioned stimulus. Furthermore, we show that MK-801, a non-competitive NMDA-R antagonist, impairs memory performance in this maze when administered right after training or just before recall but not when given before training at a dose that does not impair motor function, perception or motivation. These results suggest that the plus maze associative learning paradigm has face and construct validity and that zebrafish may become an appropriate and translationally relevant study species for the analysis of the mechanisms of vertebrate, including mammalian, learning and memory.     

Generalization in place learning across geometrically different environments is impaired by hippocampal lesions in rats

A recent study (Tommasi & Thinus-Blanc, 2004) found that when rats trained to search for food hidden in the center of a square-shaped enclosure were tested in the absence of food, they searched precisely in the center of the enclosure. Transfer tests carried out in enclosures differing in shape or size (rectangular-, large square-, and equilateral triangle-shaped) showed that rats searched in the geometric center of the novel environments. In the present work, half of those rats were lesioned bilaterally to the hippocampus and were then re-trained and re-tested in the same task, to be compared to sham rats that received the same retraining/re-test protocol. Results show that lesioned rats, although unimpaired in relearning to localize the center of the square-shaped enclosure, performed considerably worse than sham rats in the transfer tests in the enclosures differing in shape, suggesting a relevant role of the hippocampus in geometry-driven place generalization.     

Tuning the brain by learning and by stimulation of the nucleus basalis

Although it is well-established that the cerebral cortex is a substrate for learning, memory and higher cognitive functions, rather less is known about the mechanisms by which experiences are acquired and stored in the cortex. The role of the basal forebrain cholinergic system (BFCS) in learning-induced plasticity is underlined by a recent report by Kilgard and Merzenich[1]. In this article I will discuss the findings of Kilgard and Merzenich in the context of other developments in our understanding of the BFCS and its role in learning-induced plasticity. However, before the discussion I would like to provide some essential background information.     

A test of sex differences in instrumental appetitive learning by rats

A total of 48 laboratory rats, half male and half female, were trained and extinguished on instrumental bar-pressing for either 16% or 64% sucrose incentives. No consistent sex differences were found in either acquisition or variability of bar-press responding, or in extinction. The only statistically reliable difference indicated that males were more variable in responding for the 16% sucrose incentive, as measured by the range of scores over each week. It is concluded that no real sex differences in learning proficiency obtain under the experimental conditions employed, and that therefore the practice of using mainly or exclusively female rats in our laboratory is methodologically sound.     

Incentive learning and the motivational control of instrumental performance

Hungry rats were trained to press a lever and pull a chain concurrently, with one action being reinforced with a sucrose solution and the other with food pellets. In addition, in the first two experiments all animals experienced non-contingent presentations of the two incentives in the absence of the operant manipulanda while either thirsty or hungry and either before (Experiment 1A) or after (Experiment 1B) the instrumental training. When lever pressing was assessed subsequently in extinction under thirst, the animals pressed at a relatively high rate only if (1) this action had been reinforced with the sucrose solution rather than the food pellets during training and (2) they had received the non-contingent presentations of the sucrose solution and food pellets on days on which they were thirsty rather than hungry. A third experiment demonstrated that non-contingent exposure to the sucrose solution alone, but not to water under thirst was sufficient to bring about this type of motivational control of instrumental performance.     

Recollection is impaired by the modification of interpretation bias

The interpretation paradigm of cognitive-bias modification (CBM-I) was modified with instructions used in process-dissociation procedures for the purpose of investigating processes contributing to performance on the transfer task. In Experiment 1, nonanxious students were trained to interpret ambiguous situations in either a negative or benign way (or they read nonambiguous scenarios). They were then asked to respond to new ambiguous situations, in the same way as contextually similar analogues during training, or to respond differently. Benign training proactively impaired memory for negative outcomes. This effect was replicated by anxious students in Experiment 2 and discussed with respect to the assumptions underlying process-dissociation procedures and directions for future research.     

The role of the nucleus accumbens in knowing when to respond

While knowing what to expect is important, it is equally important to know when to expect it and to respond accordingly. This is apparent even in simple Pavlovian training situations in which animals learn to respond more strongly closer to reward delivery. Here we report that the nucleus accumbens core, an area well-positioned to represent information about the timing of impending rewards, plays a critical role in this timing function.     

Glucocorticoids interact with the noradrenergic arousal system in the nucleus accumbens shell to enhance memory consolidation of both appetitive and aversive taste learning

It is well established that glucocorticoid hormones strengthen the consolidation of long-term memory of emotionally arousing experiences but have little effect on memory of low-arousing experiences. Although both positive and negative emotionally arousing events tend to be well remembered, studies investigating the neural mechanism underlying glucocorticoid-induced memory enhancement focused primarily on negatively motivated training experiences. In the present study we show an involvement of glucocorticoids within the nucleus accumbens (NAc) in enhancing memory consolidation of both an appetitive and aversive form of taste learning. The specific glucocorticoid receptor (GR) agonist RU 28362 (1 or 3ng) administered bilaterally into the NAc shell, but not core, of male Sprague-Dawley rats immediately after an appetitive saccharin drinking experience dose-dependently enhanced 24-h retention of the safe taste, resulting in a facilitated attenuation of neophobia. Similarly, GR agonist infusions given into the NAc shell immediately after pairing of the saccharin taste with a malaise-inducing agent enhanced memory of this negative experience, resulting in an intensified conditioned aversion. Importantly, a suppression of noradrenergic activity within the NAc shell with the β-adrenoceptor antagonist propranolol blocked the facilitating effect of a concurrently administered GR agonist on memory consolidation in both the appetitive and aversive learning task. Thus, these findings indicate that GR activation interacts with the noradrenergic arousal system within the NAc to enhance memory consolidation of emotionally arousing training experiences regardless of valence.