Pre-training anandamide infusion within the basolateral amygdala impairs plus-maze discriminative avoidance task in rats

Endocannabinoids (eCBs) modulate a variety of brain functions via activation of the widely expressed CB1 receptor. One site of high density of this receptor is the basolateral amygdala (BLA), a structure involved in the formation of aversive memories. The activation and blockade of CB1 receptors by systemic or hippocampal drug administrations have been shown to modify memory processing. However, little is known about the role of the BLA endocannabinoid system in aversive memories. Additionally, BLA endocannabinoid transmission seems to be related to emotional states, but the relevance of these effects to memory formation is still unknown. In this study we investigated the effects of the eCB anandamide (AEA) and the CB1 antagonist/inverse agonist AM251 infused into the BLA on the acquisition of an aversive memory task, concomitantly evaluating basal anxiety levels in rats. Male rats received pre-training micro-injection of AEA, AM251 or vehicle bilaterally into the BLA, and were studied with the plus-maze discriminative avoidance task (a paradigm that allows concomitant and independent evaluation of anxiety-like behavior and the memory of an aversive task). Our results showed that AEA into the BLA before training prevented memory retrieval 24 h later, as evaluated by exploration of the aversive arm of the maze, while AM251 into the BLA did not interfere with animals' performance. In addition, AEA had no effect on anxiety-like behavior (as evaluated by open arm exploration and risk assessment), while AM251 induced an anxiogenic effect. Our data indicate an important role of BLA CB1 receptors in aversive memory formation, and suggest that this involvement is not necessarily related to a possible modulation of anxiety states.     

Acute predator stress impairs the consolidation and retrieval of hippocampus-dependent memory in male and female rats

We have studied the effects of an acute predator stress experience on spatial learning and memory in adult male and female Sprague-Dawley rats. All rats were trained to learn the location of a hidden escape platform in the radial-arm water maze (RAWM), a hippocampus-dependent spatial memory task. In the control (non-stress) condition, female rats were superior to the males in the accuracy and consistency of their spatial memory performance tested over multiple days of training. In the stress condition, rats were exposed to the cat for 30 min immediately before or after learning, or before the 24-h memory test. Predator stress dramatically increased corticosterone levels in males and females, with females exhibiting greater baseline and stress-evoked responses than males. Despite these sex differences in the overall magnitudes of corticosterone levels, there were significant sex-independent correlations involving basal and stress-evoked corticosterone levels, and memory performance. Most importantly, predator stress impaired short-term memory, as well as processes involved in memory consolidation and retrieval, in male and female rats. Overall, we have found that an intense, ethologically relevant stressor produced a largely equivalent impairment of memory in male and female rats, and sex-independent corticosterone-memory correlations. These findings may provide insight into commonalities in how traumatic stress affects the brain and memory in men and women.     

Reversal of retrieval impairment caused by retroactive interference on a two-way active avoidance task in rats

Rats were exposed to an open field with flashing light (OFL; 60-W lamp, 30 Hz, for 7 min) 2 h after training and/or 2 h before testing in a two-way active avoidance task (30 trials, 0.5-mA footshock). Post-training OFL presentation caused retroactive interference, i.e., a retrieval impairment/amnesia for the avoidance task. Pretest OFL exposure reversed the post-training OFL-induced retrieval deficit. Diazepam (2.0 mg/kg), atropine (2.0 mg/kg), and methylatropine (0.1 mg/kg) administered before post-training OFL presentation blocked the OFL amnesic effect. However, these drugs did not counteract the pretest OFL-induced recovery of retrieval. Atropine and methylatropine administered 2 h before testing to rats receiving only post-training OFL presentation canceled the OFL-interfering effect. These results suggest: (1) that the amnesic effect of post-training OFL is due to failure of retrieval of the avoidance task, (2) that the reversal of this retrieval impairment by pretest OFL exposure may involve either priming or state-dependent mechanisms, and (3) that there are different modulatory mechanisms involved in post-training and pretest OFL effects.     

Effects of sleep deprivation on free-operant avoidance

Two studies examined effects of sleep deprivation on free-operant avoidance by rats. In Experiment 1, a 5-s shock-shock (SS) interval and 20-s response-shock (RS) interval produced baseline performances, which were reestablished after each experimental manipulation. Once baselines were established, animals were exposed to 24, 48, or 96 hr of sleep deprivation and equivalent periods of home cage and food restriction as a control condition. Compared to baseline, sleep deprivation increased response rates by increasing the proportion of brief interresponse times (IRTs); response rates changed little in the control conditions. Percentage of shocks avoided did not systematically change across conditions. In Experiment 2, the RS interval was manipulated (10, 20, and 40 s), while the SS interval (5 s) and level of sleep deprivation (48 hr) were held constant. Across RS intervals, sleep deprivation increased response rates via a shift toward brief IRTs. In addition, sleep deprivation increased the percentage of shocks avoided as an inverse function of RS intervals.     

Intrahippocampal scopolamine impairs both acquisition and consolidation of contextual fear conditioning

Lesions of the dorsal hippocampus have been shown to disrupt both the acquisition and the consolidation of memories associated with contextual fear (fear of the place of conditioning), but do not affect fear conditioning to discrete cues (e.g., a tone). Blockade of central muscarinic cholinergic receptor activation results in selective acquisition deficits of contextual fear conditioning, but reportedly has little effect on consolidation. Here we show for the first time that direct infusion of the muscarinic cholinergic receptor antagonist, scopolamine, into the dorsal hippocampus produces a dose-dependent deficit in both acquisition and consolidation of contextual fear conditioning, while having no impact on simple tone conditioning.     

Reversible inactivation of the hippocampal mossy fiber synapses in mice impairs spatial learning, but neither consolidation nor memory retrieval, in the Morris navigation task

The role played by hippocampal mossy fibers in the learning and memory processes implemented in the Morris swimming navigation task has been studied in C57BL/6 mice by selective and reversible inactivation of mossy fiber synaptic fields by diethyldithiocarbamate. The functional integrity of the mossy fibers proved essential for the storage of the spatial representation on the modifiable synapses of the recurrent collaterals of the CA3 pyramidal cells, whereas it is not necessary for the consolidation and recall of spatial memories. The results suggest that mossy fibers are preferentially involved in new learning. They are consistent with the hypothesis that the hippocampal CA3 region might act as an autoassociation memory.     

Twenty-four hours of total sleep deprivation selectively impairs attentional networks

Performance decrements after sleep loss have been extensively studied and are usually attributed to generic attentional deficits. This claim, however, is based on the view of attention as a unitary construct, despite evidence that it should be considered a multidimensional cognitive ability. The aim of the present study was to evaluate the impact of one night of sleep deprivation on the efficiency of three attentional networks, defined by Posner and Raichle (1994) in anatomical and functional terms, as alerting, orienting, and executive control. Thirty participants performed the Attention Network Test at 9:00 a.m. following two different sleep conditions: baseline (a normal night of sleep) and deprivation (24 hrs of wakefulness). Results showed an overall slowing in reaction times and a significant decrease in accuracy after sleep deprivation. Sleep deprivation selectively affected the three attentional networks, given that only executive control efficacy significantly decreased after sleep deprivation. By contrast, phasic alerting and orienting showed no differences in the two sleep conditions. Thus, performance deficits following sleep deprivation do not reflect global attentional deficits.     

Perinatal linoleate deprivation impairs learning and memory in adult rats

Rats receiving polyunsaturated fatty acid (PUFA) deficient diets during the perinatal period showed in adult age undisturbed acquisition of a footshock motivated brightness discrimination task, but a significant impairment of retention. The same effects on retention were obtained in rats receiving the PUFA deficient diet in adulthood, when the behavioral parameters were investigated at the end of the dietary treatment.     

Total sleep deprivation impairs the encoding of trace-conditioned memory in the rat

Sleep may help consolidate the information of certain memories, though its benefits in the consolidation of trace-conditioned memory still remain elusive. We investigated the effect of sleep deprivation on trace learning in male wistar rats. Rats were trained for trace conditioning and the number of head entries into liquid dispenser was accounted as an outcome measure of trace-learning. For training and testing, 75 presentations of conditioned stimulus (CS) (light) and unconditioned stimulus (US) (juice) were offered in five sessions (15 presentations/session; with 5 min inter-session gap). The duration of CS and US stimuli were 15 and 20s respectively, with 5s trace delay between stimuli and 20s condition delay between each presentation. The animals were divided randomly into three groups soon after training, sleep deprived (SD) (n=8), non-SD (NSD) (n=8) and stress control (n=5) groups. The animals of NSD and control groups were left undisturbed, while SD animals were sleep deprived for 6h after training. The learning of trace-conditioned task was examined on following days. We observed that SD rats poked approximately 63% less than NSD and control groups (p<0.001) to obtain juice on testing day. Also, the NSD rats exhibited significant positive correlation in number of head entries during the training and testing days; while the SD rats showed no significant correlation. The results demonstrate that SD animals had difficulties to associate CS with US and suggest that sleep deprivation soon after training impairs the encoding of trace memory.     

Amitriptyline administered after consolidation of inhibitory avoidance does not affect memory retrieval

In Experiment 1, the effect of the administration of the antidepressant amitriptyline (30 mg/kg) for 21 days on the acquisition and consolidation of the inhibitory avoidance task was studied in male and female mice. In Experiment 2, it was evaluated whether amitriptyline administered after the consolidation of this task would block the memory retrieval. Anxiety and spontaneous activity in the elevated plus maze were also assessed. When amitriptyline was given before the training phase of inhibitory avoidance it blocked learning in males and there was a tendency in the same direction in females. However, the drug administered between training and test phases did not affect conditioning. These effects of amitriptyline seem to be independent of its actions on anxiety and locomotor activity. It may be that the effects observed are related to the therapeutic effects of antidepressants.     

Stress impairs optimal behavior in a water foraging choice task in rats

Stress is a biologically significant social–environmental factor that plays a pervasive role in influencing human and animal behaviors. While stress effects on various types of memory are well characterized, its effects on other cognitive functions are relatively unknown. Here, we investigated the effects of acute, uncontrollable stress on subsequent decision-making performance in rats, using a computer vision-based water foraging choice task. Experiencing stress significantly impaired the animals'' ability to progressively bias (but not maintain) their responses toward the larger reward when transitioning from equal to unequal reward quantities. Temporary inactivation of the amygdala during stress, however, blocked impairing effects on decision making.It is now well documented that exposure to uncontrollable stress can produce alterations in multiple brain–memory systems in humans and animals (McEwen and Sapolsky 1995; Kim and Diamond 2002; Joels et al. 2006; Shors 2006; Luethi et al. 2008). In humans, impairments in long-term, but not short-term, verbal recall tasks have been observed in people with post-traumatic stress disorder (PTSD) (Bremner et al. 1995) or Cushing''s disease (a hypercortisolemia condition) (Starkman et al. 1992) and in healthy individuals subjected to stress (Lupien et al. 1997) or exposed to stress levels of cortisol (Newcomer et al. 1994). In rodents, stress and corticosterone administration interfere with spatial and working memory (Diamond and Rose 1994; de Quervain et al. 1998; Kim et al. 2001) and potentiate aversive conditioning (Shors et al. 1992; Maier et al. 1995). Further, a number of stress-associated neurobiological changes have been identified (e.g., in hippocampus, medial prefrontal cortex, and amygdala) subserving different memory functions (Arnsten and Goldman-Rakic 1998; Kim and Yoon 1998; Vyas et al. 2003; Holmes and Wellman 2009).Although stress effects on memory have been well studied, far less is known about whether (and in what manner) stress influences other higher cognitive functions. The present study investigated the effects of acute, uncontrollable stress (60-min restraint + 60 intermittent tailshocks) on subsequent decision-making performance in rats. The stress procedure, in which animals learn that they can neither escape nor predict an aversive experience, was adapted from earlier studies (e.g., Maier and Seligman 1976; Kim et al. 1996). Decision making was assessed using an automated Figure-8-shaped maze on which rats were motivated to forage for water rewards in two different locations under equal and unequal quantity conditions (Fig. 1). In addition to behavioral stress, we examined the effects of corticosterone administration and inactivation of the amygdala during stress on decision making. Both corticosterone (a glucocorticoid hormone released in response to stress) and the amygdala (a structure crucial in defensive behavior) have been implicated in mediating neurocognitive effects of stress (McEwen and Sapolsky 1995; Kim and Diamond 2002).Open in a separate windowFigure 1.Decision-making task. Thirst-motivated rats were trained to forage for water on an automated Figure-8-shaped maze. A computer algorithm controlled the raising and lowering of four gates (represented by rectangles), the delivery of water (blue circles), and tracking of the animal''s location on the maze. During the baseline days, both left and right sides of the maze presented 0.04 mL of water at 80% probability (equal reward trials). Following each trial (a left or right loop), the animal returned to the center bridge to start the next trial (40 trials per day). During the bias test days, one side of the maze (counterbalanced) offered 0.12 mL of water at 80% probability, while the other side continued to present 0.04 mL of water at 80% probability (unequal reward trials). Animals received either stress, CORT injections, or neither (for group details, see text).Experimentally naïve male Charles River Sprague–Dawley rats (initially weighing 275–300 gm) were singly housed and maintained on a reverse 12-h light-dark cycle (lights on at 19:00 h). After 7 d of acclimation and for the duration of the experiment, daily water access was restricted to maintain approximately 85% of the animal''s normal body weight. Food was available ad libitum throughout the experiment. All experiments were conducted during the dark phase of the cycle and in strict compliance with the University of Washington Institutional Animal Care and Use Committee guidelines. Under anesthesia (30 mg/kg ketamine and 2.5 mg/kg xylazine, i.p.) amygdala (AMYG) animals were chronically implanted with 26-gauge guide cannulae (Plastics One) bilaterally into the amygdala (from bregma: anteroposterior, −2.3 mm; mediolateral, ±5 mm; dorsoventral, −7.7 to 8.0 mm). During 10–15 d of postoperative recovery, each dummy cannula was removed and replaced with a clean one.Muscimol free base (Sigma-Aldrich), dissolved in artificial cerebrospinal fluid (10 mM at pH ∼7.4), was microinfused into the amygdala (bilaterally) via 33-gauge infusion cannulae that protruded 1 mm beyond the guide cannulae (cf. Kim et al. 2005). An infusion volume of 0.3 μL (per side) was delivered using a Harvard PHD2000 syringe pump (Harvard Apparatus) over the course of 3 min. Animals were returned to their home cages for 30 min before undergoing the stress procedure. We based the timing of inactivation on previous findings that pre-stress but not immediate post-stress inactivation of the amygdala interferes with stress effects on hippocampal long-term potentiation (LTP) and spatial memory (Kim et al. 2005). Based on studies that examined ³H-muscimol spreading (Krupa et al. 1993; Arikan et al. 2002) in the cerebellum, in which 1 μL diffused a radius of 1.6–2.0 mm, it was estimated that 0.3 μL of muscimol would spread within a radius of approximately 0.5–0.7 mm from the infusion needle tip. Hence, it is likely that infused muscimol would have diffused to the central, lateral, and basal nuclei of the amygdala and possibly to portions of adjacent neighboring structures. The BODIPY TMR-X muscimol conjugate (Invitrogen) was infused in the same manner as muscimol free base (cf. Allen et al. 2008) to image the spread of reversible amygdalar inactivation.Corticosterone (CORT) animals received three daily subcutaneous injections of 3 mg/kg corticosterone (suspended in sesame oil; Sigma-Aldrich) 30 min prior to bias testing.Rats undergoing stress were restrained in Plexiglas tubes and presented with 60 tailshocks (1-mA intensity, 1-sec duration, 5- to 115-sec variable intershock interval) for 60 min (Kim et al. 2005). Animals were divided into four groups: control, stress, amygdalar inactivation plus stress (AMYG), and daily corticosterone (CORT).Following 2 d of habituation (to transport, maze, and room ambiance), all animals underwent successive shaping and testing phases. The dimensions and automatic features of the Figure-8 maze have been described previously (Pedigo et al. 2006; Yoon et al. 2008); for details, see Supplemental material. During shaping, each rat is placed into the center runway with all four gates in the up position (Supplemental Fig. S1). After 3 sec, the front and one of the side gates drop, until the rat on its own volition moves out of the center and onto the open side runway. At this point, the lowered front and side gates rise (to prevent the rat from going backward), water is delivered to both the open arm and the center spout, and the back gate drops. The rat consumes water from the open arm, and a new trial begins when he returns to and consumes water from the center arm. There was a 3-sec delay between each trial. During shaping, left and right choices were thus forced choices and were presented in a pseudorandom pattern, such that there was an equal number of both in a complete session (40 laps). Rats underwent shaping once daily until they met predetermined criteria: completion of 40 laps in less than 30 min, and less than five back edge errors (i.e., after making a choice, running up the opposite arm instead of going back to the center arm). The automated program controlled the gates and water delivery, according to the rat''s position on the maze.During baseline testing (Supplemental Fig. S2), the rewards dispensed on left and right arms were equal in both volume (0.04 mL) and probability (0.8). Each rat remained on baseline testing until he demonstrated a stable left/right choice pattern across three consecutive days. If a slight preference to one side was present, the opposite arm would be the increased reward side when bias testing commenced.Stress and AMYG rats were exposed to restraint + tailshock stress 1 d before their first bias test, and CORT rats were given corticosterone injections 30 min prior to each bias test. During bias testing (Supplemental Fig. S3), the reward value on one side was tripled in volume (0.12 mL) while the value for the other side remained constant (0.04 mL). Control, AMYG, and CORT animals were given three consecutive days of bias tests, while stress animals underwent six consecutive days of bias tests.At the completion of behavioral testing, animals were overdosed with urethane and perfused intracardially with 0.9% saline followed by 10% buffered formalin. The brains were removed and stored in 10% formalin overnight and then kept in 30% sucrose solution until they sank. Transverse sections (50 μm) were taken through the extent of the cannulae tract, mounted on gelatin-coated slides, and stained with cresyl violet to verify cannulae placements.The visit number to the left (or right) side of the maze for baseline and bias days were normalized to the mean left (or right) visits across the three baseline days. Statistical comparisons between groups were examined using ANOVA. For a significant difference (P < 0.05), post-hoc comparisons were performed using Tukey''s honestly significant difference test.Thirst-motivated rats readily learned to forage for water on the maze, and when left and right sides of the maze provided the same quantity (0.04 mL) and probability (80%) of water, the animals made comparable numbers of left and right visits (during 40 laps daily) that were stable across three baseline days (Fig. 2A). The 80% probability (a partial reinforcement schedule) was used so that animals frequently explored both sides of the maze. After animals demonstrated stable baseline choices, the volume of water on one side was tripled (0.12 mL at 80%), while the other side remained constant (0.04 mL at 80%). Overall, bias performance (choice of the larger reward) increased across the first three bias test days (repeated-measures ANOVA; main effect of day, F_(2,54) = 78.16, P < 0.001). However, the four groups differentially increased their bias across days (group × day interaction, F_(6,54) = 4.14, P < 0.01). Specifically, stressed rats displayed a significantly slower rate of bias toward the larger reward than did the controls (P < 0.01, Tukey). The stressed rats did ultimately develop a bias compared with their baseline choices (F_(6,62) = 8.44, P < 0.001). This bias was first significant on the fourth day (P < 0.05, Dunnett t-test). However, even after 6 d of bias testing, their bias (127 ± 5.7%, mean ± SEM) did not reach the level of controls'' third day bias (182 ± 12.2%). Unlike the behavioral stress group, however, animals that received three daily corticosterone injections (3 mg/kg, subcutaneously) prior to testing chose the larger reward side more frequently (173 ± 6.7%, bias day 3) and did not differ from the control group (P > 0.7, Tukey). When the amygdalae were inactivated during stress (Fig. 3), these animals behaved like controls (P > 0.7, Tukey) and increased their visit frequency toward the larger reward side of the maze (174 ± 13.0%, bias day 3) (Fig. 2A). Although control, CORT, and AMYG animals developed strong bias responses, they did not exclusively visit the larger reward side of the maze because on 20% of trials they did not receive a reward.Open in a separate windowFigure 2.Stress effects on decision making. (A) All groups of animals showed comparable visits to left and right sides of the maze during the three baseline days. When transitioning from equal to unequal reward trials, stressed rats (n = 7) displayed an impaired ability to bias their responses toward the larger reward side compared with control (n = 10), AMYG (n = 7), and CORT (n = 7) rats (P = 0.002, group × bias day interaction). (B) Example visit maps of a control rat during baseline and bias days (40 laps each).Open in a separate windowFigure 3.(Top) Histological reconstruction of injection cannulae placement tips in the amygdala. (Bottom) A photomicrograph of fluorophore-conjugated muscimol (0.3 μL over 3 min) spread in the amygdala. The red fluorescence is overlaid with a dark field image.We then examined whether stress might have produced alterations in motor, motivation, and reference memory performances that hindered the animals'' ability to bias their responses toward the larger reward. The latency to complete 40 laps of the first bias test session (Supplemental Fig. S4A) showed a trend of stress animals completing the bias test faster than the other three groups, but this group × day interaction was not significant (F_(6,54) = 1.89, P > 0.05). Stress also did not impair the animals'' reference memory of the maze (Supplemental Fig. S4B). That is, after making a left or right visit, stressed animals readily re-entered the center runway to start the next trial (one-way ANOVA; average baseline and first three bias days, F_(3,41) = 0.20, P > 0.8), whereas control, CORT, and AMYG animals displayed an increased propensity to investigate the other side before re-entering the center, particularly as bias testing progressed (repeated-measures ANOVA; main effect of day, F_(2,54) = 4.84, P < 0.05).Our results indicate that rats clearly demonstrate the capacity to change their foraging behavior to acquire a larger water reward when transitioning from equal to unequal quantities, and that such behavioral flexibility is vulnerable to acute, uncontrollable stress. Specifically, rats that experienced 1 h of restraint stress + 60 intermittent tailshocks were significantly impaired in biasing their responses toward the side of the maze with a larger quantity of water. This effect on bias was not due to any lingering post-stress motivational or motor effects, as stress did not increase the latency to complete the bias test. Daily corticosterone injections did not interfere with this task, indicating that corticosterone elevation per se cannot reproduce behavioral stress effects on behavioral flexibility. However, similar to previous stress–memory studies (Kim et al. 2001; Waddell et al. 2008), amygdalar inactivation during stress effectively blocked this effect. This suggests that the amygdala plays a crucial role in mediating stress effects across different cognitive domains.Although stress altered the rats'' behavior in our choice-based task, it remains unclear precisely which neural and cognitive systems were affected. For instance, the impairment of behavioral flexibility might be an indirect consequence of stress effects on hippocampal memory functioning. The stress paradigm used here is known to impede LTP in the CA1 hippocampus and hinder spatial memory (Foy et al. 1987; Kim et al. 2001). However, corticosterone, which also impedes LTP (McEwen and Sapolsky 1995) and spatial memory when administered 30 min before testing (de Quervain et al. 1998), did not impair behavioral flexibility. The impairment to choose the larger reward may be due to stress effects on working memory, such that the rats cannot remember (and thus learn) that one reward is larger. However, if true, the stress-induced memory impairment is unusually persistent in our task: Rats were affected through at least 6 d beyond the stress exposure. Because acquisition and retrieval of information are integral components of decision making, the contribution of stress-associated changes in learning and memory cannot be fully excluded. Another possibility is that stressed rats are more likely to use habitual rather than flexible strategies, even when a change in behavior may be optimal. Consistent with this explanation are recent findings that chronic stress exposure increases habit-based responding, with corresponding atrophy and hypertrophy of goal-directed and habit-based neural circuitry, respectively (Dias-Ferreira et al. 2009). The reliance on habit memory is also increased following anxiogenic drug infusions into the amygdala (Elliott and Packard 2008), which further implicates amygdalar modulatory activity during and after stress exposure in decreasing flexible behavior. If the stress experience did increase perseverative choice behavior, this may partially explain previously observed associations between distress and perseveration in humans (Robinson et al. 2006). Alternately, stress may have disrupted the reward circuitry and impaired the ability to discriminate between the two reward values from the two side arms, in which case stress effects on a dopamine-related reward circuit (Schultz et al. 1997) should be explored. However, this cannot be the sole explanation because post-bias stress did not affect the animals'' bias behavior toward the larger reward (Supplemental Table S1), and nonstressed rats resume equal arm visits when rewards are returned to baseline values (data not shown).The present findings reveal that a single exposure to an acutely stressful experience is enough to affect an animal''s behavior on a simple forging task for several days. There is accumulating evidence that exposure to stress increases amygdala and decreases prefrontal activity in both humans and animals (for review, see Arnsten 2009) and that even a single stress exposure alters cellular morphology in prefrontal cortex (Izquierdo et al. 2006). This shift in neural activity may promote the use of one cognitive strategy over another (e.g., habitual versus flexible). To address this, future studies need to investigate brain structures implicated in decision making, including the prefrontal and the parietal cortices (Gold and Shadlen 2007; Lee 2008), for their susceptibility to stress. Regardless, the present findings, to our knowledge, provide the first direct evidence that acute uncontrollable stress persistently impairs decision-making performance in animals and that this effect is dependent upon amygdalar activity during stress.     

Hyperthermia impairs retrieval of an overtrained spatial task in the Morris water maze

Fifteen rats were trained to learn the location of a spatially fixed platform hidden in a Morris water maze (40 +/- 2 degrees C). Then retention of the spatial task was assessed immediately after raising core body temperature (Tc) to 42 or 40 degrees C or stabilizing at 37 degrees C (the normothermic control). The hyperthermic treatment order was counterbalanced according to a Latin-square design. Hyperthermia at 42 degrees C Tc significantly impaired spatial performance. Hyperthermic animals were cooled to normothermia (Tc = 37 degrees C) and spatial performance was tested again approximately 30 min later. Cooling resulted in a complete recovery of spatial performance. These results demonstrate that hyperthermia-induced amnesia can be obtained on an overtrained spatial-mapping strategy and cooling to normothermia initiates recovery of spatial performance.     

Four hours of paradoxical sleep deprivation impairs alternation performance in a water maze in the rat

Sixteen rats were deprived of paradoxical sleep (PS) for 4 h using the "flower pot" technique and 16 other served as yoked controls. PS-deprived and control rats then had to learn a water maze using either a standard allocentric configuration (i.e., finding the submerged platform using external cues; n = 6/group) or an alternation version (goal platform alternating between two locations; n 10/group). Rats were submitted to six trials with a cutoff time of 60 s and an intertrial interval of 5 min. Criterion was set as two consecutive successful completions. PS-deprived rats made more quadrant entries and took more time to reach criterion on the alternation task than control rats while both groups were equal on the allocentric task. Based on lesion studies (Ethier et al., this issue) we propose that tasks that require an intact medial prefrontal cortex are particularly sensitive to PS deprivation.     

Inhibition of monoADP-ribosylation prevents long-term memory consolidation of a single-trial passive avoidance task in the day-old chick

The cytosolic posttranslational protein-modifying mechanism of monoADP-ribosylation has been implicated in long-term potentiation, a synaptic model of memory formation. The current study investigated the effect of inhibiting mono(ADP-ribosyl) transferase on memory for the passive avoidance task in day-old chicks (white Leghorn-black Australorp). Various doses of novobiocin or menadione sodium bisulfite were administered intracranially at different times before or after training. Control chicks were administered saline at matched times. Novobiocin (650 microM) or menadione sodium bisulfite (250 microM) administered between 5.0 min pretraining and 2.5 min posttraining was found to cause a persistent loss of retention from 120 min posttraining. These data provide the first demonstration that monoADP-ribosylation is required for the maintenance of long-term memory. Furthermore, the temporal characteristics of the memory loss caused by monoADP-ribosylation inhibition appears to exclude this mechanism as a downstream effect of the well-established nitric oxide activity previously shown to occur within 40 min of passive avoidance training.