Reversible disconnection of the hippocampal-prelimbic cortical circuit impairs spatial learning but not passive avoidance learning in rats

Wistar rats, treated with the GABA(A) receptor agonist muscimol, were used to investigate the role of the hippocampal-prelimbic cortical (Hip-PLC) circuit in spatial learning in the Morris water maze task, and in passive avoidance learning in the step-through task. In the water maze task, animals were trained for three consecutive days and tested 24 h after the end of training. In the step-through task, the animals were trained once and tested 24h after training. On the training days, daily infusion of muscimol (0.5 microg/0.25 microl) was given (1) bilaterally to the ventral hippocampus (vHip), (2) bilaterally to the prelimbic cortex (PLC), (3) to the unilateral vHip and the ipsilateral PLC, or (4) for disconnecting the Hip-PLC circuit, to both the unilateral vHip and the contralateral PLC 30 min before training. The results showed that inhibition of the vHip resulted in disruption of performance in both tasks. Inhibition of the PLC produced impaired water maze performance, but had no effect on the step-through task. Disconnection of the Hip-PLC circuit produced similar effects to PLC inhibition. However, simultaneous inhibition of the unilateral vHip and the ipsilateral PLC had little effect on performance of the water maze task. The results suggested that spatial learning depends on the Hip-PLC circuit, whereas passive avoidance learning is independent of this circuit.     

Functional recovery of skilled forelimb use in rats obliged to use the impaired limb after grafting of the frontal cortex lesion with homotopic fetal cortex

The long-term effect of transplanting embryonic frontal cortex into a unilateral frontal cortex lesion has been studied in adult rats. Before surgery, activity in an open field, muscular strength of both forelimbs, and performance in a paw-reaching-for-food task were scored in 26 rats. In 21 animals a unilateral cortex lesion was then made in the forelimb motor area of the hemisphere contralateral to the preferred paw in the paw-reaching-for-food task, while the other 5 animals were sham-operated. On retesting, the lesion animals changed the preferred paw. A solid homotopic transplant of embryonic tissue (embryonic day 17) was then placed in the lesion cavity in 11 of the lesion rats. Three months later neither lesion alone nor lesion plus transplantation affected open field behavior and muscular strength, but the lesion permanently affected performance in the paw-reaching-for-food task, as shown by a change of preferred paw and a functional deficit in the paw contralateral to the lesion. Transplantation ameliorated the deficits caused by the lesion, but this was only evident when animals were forced to reach with the paw contralateral to the lesion plus transplant. The behavioral results were independent of the size of the lesion and graft. Connections between graft and host tissue were studied by means of the fluorescent tracer 1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate (DiI). A dense array of labeled fibers was found in the host cortex adjacent to the transplant. The results suggest that functional recovery depends on grafting but is only evident when the animal is obliged to use the affected limb.     

Sensorimotor responsiveness in rats with unilateral superior collicular and amygdaloid lesions.

Large unilateral lesions of superior colliculus, but not amygdala, result in strong ipsiversive progression tendencies and permanent neglects of visual, auditory, and whisker-touch stimuli presented on the contralateral side of the body. Combined collicular-amygdaloid lesions also yield circling behaviors and multimodal neglects that are completely independent of the order or laterality of the amygdaloid lesion. Rats with colliculectomy either neglect or else turn away from pinches of the contralateral ear and forepaw. Subjects with combined lesions display more crossed orientations, and this tendency is greatly potentiated by ipsilateral lesion placement. The nature and time course of the crossed response in rats with sensory neglects is reminiscent of the clinical syndrome described as alloaesthesia or as contralateral sensory displacement.     

Interaction between the basolateral amygdala and dorsal hippocampus is critical for cocaine memory reconsolidation and subsequent drug context-induced cocaine-seeking behavior in rats

Contextual stimulus control over instrumental drug-seeking behavior relies on the reconsolidation of context-response-drug associative memories into long-term memory storage following retrieval-induced destabilization. According to previous studies, the basolateral amygdala (BLA) and dorsal hippocampus (DH) regulate cocaine-related memory reconsolidation; however, it is not known whether these brain regions interact or independently control this phenomenon. To investigate this question, rats were trained to lever press for cocaine reinforcement in a distinct environmental context followed by extinction training in a different context. Rats were then briefly re-exposed to the cocaine-paired context to destabilize cocaine-related memories, or they were exposed to an unpaired context. Immediately thereafter, the rats received unilateral microinfusions of anisomycin (ANI) into the BLA plus baclofen/muscimol (B/M) into the contralateral (BLA/DH disconnection) or ipsilateral DH, or they received contralateral or ipsilateral microinfusions of vehicle. They then remained in their home cages overnight or for 21 d, followed by additional extinction training and a test of cocaine-seeking behavior (nonreinforced active lever responding). BLA/DH disconnection following re-exposure to the cocaine-paired context, but not the unpaired context, impaired subsequent drug context-induced cocaine-seeking behavior relative to vehicle or ipsilateral ANI + B/M treatment. Prolonged home cage stay elicited a time-dependent increase, or incubation, of drug-context-induced cocaine-seeking behavior, and BLA/DH disconnection inhibited this incubation effect despite some recovery of cocaine-seeking behavior. Thus, the BLA and DH interact to regulate the reconsolidation of cocaine-related associative memories, thereby facilitating the ability of drug-paired contexts to trigger cocaine-seeking behavior and contributing to the incubation of cocaine-seeking behavior.     

A qualitative aspect of learning-sensitive open field ambulation in mice

Maze training was shown to affect qualitative aspects of the learning-sensitive open-field (OF) ambulation in male Swiss albino mice. The changes in thigmotactic (wall-seeking) behaviour were most pronounced when significant signs of learning occurred, usually on the third day of training. Also daily changes in thigmotaxis from before to after training were found. Mice not subjected to maze training exhibited a different thigmotactic pattern of behaviour. The present series of experiments indicated that separate starting-points (centre or wall) influenced the thigmotactic behaviour as opposed to the shape of the OF (circular or square).     

Development of place navigation in rats from weaning to puberty

Young hooded rats were trained to escape onto a hidden platform after swimming in a pool of opaque water. Subjects 21, 28, 35, 42, and 64 days of age on the first training day were given 28 trials on 5 consecutive days. Half of the rats were required to localize the platform in relation to external room cues only ("place only" condition) and the other half were helped by the presence of a visible cue on the platform ("cue + place" condition). A deficiency in place navigation was observed in the 21- and 28-day groups; they showed slow escape and took circuitous routes more often than older rats. This deficiency was related to a poor spatial bias toward the training position when the subjects were allowed to swim for 30 s in the absence of the platform, at the end of the 28-trial training period (probe trial). The 35-day group showed adult-like learning ability in both training conditions, but failed to show searching behavior during the probe trial after having been trained in the presence of the proximal cue. Only rats older than 40 days showed typical adult behavior such as swimming directly toward the platform from any starting position and localized searching around the absent platform's position during the probe trial, no matter what the training conditions were. These results suggest that central nervous system structures responsible for place learning in the rat are functional from around 32 days of age, but fail to trigger searching behavior following cued training before the sixth week.     

Activation and survival of immature neurons in the dentate gyrus with spatial memory is dependent on time of exposure to spatial learning and age of cells at examination

Neurogenesis continues to occur throughout life in the dentate gyrus of the hippocampus and may be related to hippocampus-dependent learning. We have recently reported that there is an enhancement of neurogenesis in the hippocampus only when BrdU is administered 6 days prior to starting spatial training but not when training started either 1 day or 11 days following BrdU administration. In that study, all rats were perfused on day 16 after BrdU injection in order to compare cells of the same age (i.e. 16 day old cells) and thus the survival time after learning was different between groups. This study was designed to address whether the amount of time that passed following training could also contribute to the effects of spatial learning on hippocampal neurogenesis and whether there was differential new neuron activation in response to spatial learning that depended on the age of new cells at the time of spatial learning. Here we tested whether a survival period of 5 days following spatial learning at either 1-5, 6-10 or 11-15 days following BrdU administration would alter cell survival and/or activation of new neurons. Our results indicate that 5 days after training in the Morris water task cell survival is unaltered by training on days 1-5, increased by training at days 6-10 and decreased when training occurs on days 11-15. Furthermore spatial learners trained on days 6-10 or 11-15 show greater activation of new neurons compared to cue-trained rats during a probe trial 5 days after training. In addition, rats trained on the spatial task on days 11-15 had a greater number of activated new neurons compared to rats trained on the spatial task on days 6-10. These results suggest there is a gradual removal of older BrdU-labeled new neurons following spatial learning perhaps due to a competitive interaction with a population of younger BrdU-labeled new neurons.     

A conditioned reinforcer maintained by temporal association with the termination of shock

Two experiments were conducted to determine whether a stimulus can be established as a positive conditioned reinforcer by associating it with the termination of shock, but without training the animal to make any response in its presence. In the first, six rats were conditioned to press a bar to terminate shock on a variable ratio schedule; white noise was then substituted as the immediate consequence, with the shock terminating 30 sec after the last press in its presence. It was found that the rate of pressing in the absence of noise depended on the contingency between the pressing and the noise. The second experiment sought to determine whether the difference in rates before and after the onset of the noise was due to the reinforcement of prior responding by the onset of the noise or to the suppression of subsequent responding by differential reinforcement of competing behavior. Six more rats were trained in the same manner, but with shock terminating 30 sec after the onset of the noise, regardless of what the animal did in its presence. Again the rate was higher before the onset of the noise, indicating that pressing was indeed maintained by the noise as a conditioned reinforcer.     

Brief reaching training with “sticky mittens” in preterm infants: Randomized controlled trial

ObjectiveTo examine whether a brief reaching training with sticky mittens was effective to improve reaching behavior in newly reaching preterm infants.MethodsIn this randomized controlled trial, twenty four 5-month-old (±16-week-old corrected age) preterm infants were randomly allocated into experimental or control groups. Infants were assessed three times in a single session: pretraining (immediately before training), posttraining (immediately after training), and retention (4 min after the posttraining). During training, infants in the experimental group wore open fingers Velcro covered mittens. Training consisted of one 4-minute session of stimulated reaching using Velcro covered toys. Controls did not receive the training. During assessments, infants were placed in a baby chair and toys without Velcro were offered at their midline for 2 min. Number of total reaches, proximal adjustments and distal adjustments of reaching were primary outcomes. Grasping was a secondary outcome.ResultsGroups were similar in the pretraining. In the posttraining, trained infants performed greater amount of total reaches and bimanual reaches than untrained infants. Greater amount of bimanual reaches in trained infants was maintained in the retention test. Distal adjustments and grasping outcome were not influenced by the training.ConclusionsA brief-term training with open fingers sticky mittens benefited reaching behavior and favored retention of increased bimanual reaches in newly reaching late preterm infants. However, it was not sufficient to influence hand openness and early grasping.     

Metyrapone reveals that previous chronic stress differentially impairs hippocampal-dependent memory

Chronic stress facilitates fear conditioning in rats with hippocampal neuronal atrophy and in rats in which the atrophy is prevented with tianeptine, a serotonin re-uptake enhancer. The purpose of this study was to determine whether the lack of dissociation between fear conditioning performance and hippocampal integrity was masked by the presence of endogenous corticosteroids during training. As in previous studies, rats were stressed by daily restraint (6 h/day for 21 days), trained in the conditioning chamber (day 23), and then assessed for conditioned fear (day 25) at a time when hippocampal dendritic atrophy persists. On the training day, half of the control and stressed rats were. injected with metyrapone to reduce corticosterone release. Two hours later, two paired or unpaired presentations of tone and footshock were delivered. Although metyrapone reduced conditioned fear in all rats, only stressed rats showed dissociated fear conditioning (i.e. tone conditioning was reduced while contextual conditioning was eliminated). Chronically stressed rats, regardless of metyrapone treatment displayed more rearing in the open field when tested immediately after the completion of fear conditioning. These data support the hypothesis that increased emotionality and enhanced fear conditioning exhibited by chronically stressed rats maybe due to endogenous corticosterone secretion at the time of fear conditioned training. Moreover,these data suggest that chronic stress impairs hippocampal-dependent processes more robustly than hippocampal-independent processes after metyrapone to reduce corticosterone secretion during aversive training.     

Unilateral olfactory conditioning in 6-day-old rat pups

The potential that early olfactory learning might be laterally organized in the brain was investigated in 6-day-old rats. This hypothesis is based on the finding that the commissural systems that subserve bilateral olfactory communication do not mature until the second week of postnatal life. Pups were trained with pairings of cedar odor and intraoral infusions of milk while one nostril was occluded. Animals expressed a conditioned orientation towards cedar if tested with the trained nostril open. No such conditioning was observed if the untrained nostril was open during testing. Further, when individual pups received cedar odor/milk pairings with one nostril open and orange odor/milk pairings with the other open, they expressed a conditioned preference for orange when tested with the orange-trained nostril open, and a preference for cedar when tested with the cedar-trained nostril open. Classically conditioned oral responses (mouthing) also appeared to be lateralized. However, no such unilateral conditioning occurred with respect to behavioral activation, which is also conditioned in this paradigm. Increases in activity to the odor CS were observed regardless of whether the trained or untrained nostril was open during testing. These results suggest that in developing rodents, olfactory memories may be partly represented in structures that can be unilaterally accessed during training and testing. They provide a starting point for isolation of neural substrates of the olfactory conditioning process.     

Maze-running and thigmotaxis in mice: Applicability of models across the sexes

Both male (n = 244) and female (n = 223) Swiss albino mice displayed a U-shaped pattern of thigmotaxis (wall-seeking) in a circular open-field (OF) before and after a daily maze-running. A significant increase of mice's tendency to occupy the inner partitions of the OF after the maze trial on the last three out of five days of training was observed. When factor analyzed, the variables indicating the wall-seeking behaviour of both sexes were divided into two factors interpreted as "spontaneous" in conjunction with the first three to four maze trials and as "learning-related thigmotactic behaviour" during the subsequent trials in the course of the learning process.     

Long-term sensitization training primes Aplysia for further learning

Repetitive, unilateral stimulation of Aplysia induces long-term sensitization (LTS) of ipsilaterally elicited siphon-withdrawal responses. Whereas some morphological effects of training appear only on ipsilateral sensory neurons, others appear bilaterally. We tested the possibility that contralateral morphological modifications may have functional significance. Therefore, we examined whether LTS training primes subsequent sensitization. Twenty-four hours after LTS training the effects of brief shock treatment (BST) were examined. BST failed to sensitize animals that had previously received either 4-d control treatment or 4-d ipsilateral LTS training. In contrast, BST did sensitize animals that had previously received 4-d contralateral LTS training, suggesting the presence of a latent trace that primes the animal for further learning.     

A further experiment on the discrimination of open and closed shapes by rats

In a previous experiment on the visual discrimination of open and closed shapes by rats (Sutherland, Carr, and Mackintosh, 1962) it was found that (1) at the beginning of training the animals exhibited a strong preference for the more open shape; (2) shapes differing in their horizontal projections only were more discriminable than shapes differing in their vertical projections only. The present experiment is a replication of the earlier one using different shapes and different apparatus. The first result was fully confirmed and some confirmation was obtained for the second, although late in training the differences between groups trained with shapes differing on opposite projections were not statistically significant.     

Naloxone administration impairs autoshaped learning

Effects of naloxone on acquisition of autoshaped behavior were investigated. Rats deprived to 85% of free-feeding weights were trained to touch a retractable lever; delivery of a food pellet occurred on every trial following lever retraction. The lever was retracted immediately if a touch occurred within 15 s, or automatically after 15 s. Analyses were conducted on number and latencies of touches of the extended lever, nose-pokes (touches) directed at the retracted lever during intertrial intervals (a measure less constrained by ceiling effects than extended lever touches), and unconditioned exploratory rearing activity, measured as touches of a metal strip mounted above the grid floor of the apparatus. In an initial experiment, male Sprague-Dawley rats were given saline or naloxone (2.0 mg/kg, ip) 5 min before a training session of 12 trials. Two days later they were tested, in the absence of drug, in a session of 36 (three blocks of 12) trials. Naloxone depressed training levels of lever responding, in addition to slowing acquisition rate. No effect of naloxone was observed on rearing activity. Previous work showed that injection of saline 5 min before behavioral testing increases the rate of autoshaping compared to injections 30 min before (Messing & Sparber, 1984). Thus, effects of naloxone on acquisition of lever-directed behaviors may have been confounded by behavioral depressant effects and/or by an injection effect such a short time before testing. In a second experiment naloxone (0.5 or 2.0 mg/kg) was injected after five of seven training sessions (12 trials each) to male and female rats. A 6-s delay of reinforcement was inserted between lever retraction and food delivery, slowing acquisition rates and providing the opportunity to test the effects of naloxone throughout a multiple-session task. The low dose retarded acquisition of extended lever touching in both sexes; both doses retarded acquisition of interim lever touching in males. Thus, in some circumstances, post-training naloxone administration may impair learning. The results support the notion that low doses of naloxone may have agonist activity.     

Pairings of a Circular Alley with Apomorphine Produce Conditioned Hyperactivity

During training trials, some rats were injected with apomorphine as the unconditioned stimulus immediately before (Group Forward) or 15 min before (Group Backward) exposure to the conditioned stimulus, a circular alley where locomotor activity was measured for 30 min. Rats in the control condition (Group Unpaired) were not injected until 15 min after their removal from the alley. When the rats were later tested in the circular alley without prior administration of the drug, both Groups Forward and Backward were more active than Group Unpaired and thus showed conditioned hyperactivity. During a final test in which all the rats were injected with apomorphine prior to placement in the circular alley, Groups Forward and Backward showed greater apomorphine-induced hyperactivity than Group Unpaired. Although Group Forward was more active than Group Backward during both tests, these differences were not significant. These findings provide further evidence for the modulation of drug effects by Pavlovian conditioning.     

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.     

Accumbens shell AMPA receptors mediate expression of extinguished reward seeking through interactions with basolateral amygdala

Extinction is the reduction in drug seeking when the contingency between drug seeking behavior and the delivery of drug reward is broken. Here, we investigated a role for the nucleus accumbens shell (AcbSh). Rats were trained to respond for 4% (v/v) alcoholic beer in one context (Context A) followed by extinction in a second context (Context B). Rats were subsequently tested in the training context, A (ABA), or the extinction context, B (ABB). Pre-test injections of the glutamate AMPA receptor antagonist, NBQX (1 μg) into AcbSh had no effect on renewal of alcoholic beer seeking when rats were returned to the training context (ABA). However, NBQX increased responding when rats were tested in the extinction context (ABB). In a second experiment, rats received training, extinction, and test in the same context. Pre-test injections of NBQX (0, 0.3, and 1 μg) into the AcbSh dose-dependently attenuated expression of extinction. We also found that NBQX in the AcbSh had no effect on initial acquisition of extinction or the motivation to respond for reward as measured by break point on a progressive ratio schedule. Finally, we show that pharmacological disconnection of a basolateral amygdala (BLA) → AcbSh pathway via NBQX in AcbSh combined with reversible inactivation of the contralateral BLA attenuates expression of extinction. Together, these results suggest that AcbSh AMPA receptors mediate expression of extinguished reward seeking through glutamatergic inputs from the BLA.     

Medial auditory thalamus inactivation prevents acquisition and retention of eyeblink conditioning

The auditory conditioned stimulus (CS) pathway that is necessary for delay eyeblink conditioning was investigated using reversible inactivation of the medial auditory thalamic nuclei (MATN) consisting of the medial division of the medial geniculate (MGm), suprageniculate (SG), and posterior intralaminar nucleus (PIN). Rats were given saline or muscimol infusions into the MATN contralateral to the trained eye before each of four conditioning sessions with an auditory CS. Rats were then given four additional sessions without infusions to assess savings from the initial training. All rats were then given a retention test with a muscimol infusion followed by a recovery session. Muscimol infusions through cannula placements within 0.5 mm of the MGm prevented acquisition of eyeblink conditioned responses (CRs) and also blocked CR retention. Cannula placements more than 0.5 mm from the MATN did not completely block CR acquisition and had a partial effect on CR retention. The primary and secondary effects of MATN inactivation were examined with 2-deoxy-glucose (2-DG) autoradiography. Differences in 2-DG uptake in the auditory thalamus were consistent with the cannula placements and behavioral results. Differences in 2-DG uptake were found between groups in the ipsilateral auditory cortex, basilar pontine nuclei, and inferior colliculus. Results from this experiment indicate that the MATN contralateral to the trained eye and its projection to the pontine nuclei are necessary for acquisition and retention of eyeblink CRs to an auditory CS.     

Individual differences in anxiety trait are related to spatial learning abilities and hippocampal expression of mineralocorticoid receptors

Although high levels of anxiety might be expected to negatively influence learning and memory, it remains to be shown whether individual differences in anxiety may influence spatial learning and memory in outbred rat populations. We have studied this possibility in male Wistar rats whose levels of anxiety were first characterized as either high (HA) or low (LA) according to their behavior in the elevated plus maze or in the open field test. Subsequently, their performance in the Morris water maze was studied, a task dependent on hippocampal activity. Interestingly, LA rats showed a faster acquisition and better memory in the water maze when compared to HA rats. Indeed, this difference in performance could mainly be attributed to the increase in thigmotactic behavior (swimming in circles close to the maze walls) displayed by HA rats during spatial navigation. Glucocorticoids are known to affect the state of anxiety and the hippocampus is the main target of glucocorticoids in the brain. Hence, we investigated whether the hippocampal expression of the two classical corticosteroid receptors, mineralocorticoid (MR) and glucocorticoid (GR) differed in the two groups of rats. We found that LA rats displayed higher hippocampal expression of MR but not GR than HA rats. Indeed, the expression levels for these receptors were positively correlated with the amount of time spent by the animals in the open arms of the elevated plus maze. Moreover, we present evidence that the levels of anxiety quantified in the first stages of our study constitute a trait rather than a state. Taken together, this study has generated evidence of a close interaction between the anxiety trait, hippocampal MR expression and the learning abilities of individuals in stressful spatial orientation tasks.