Differential effects of acute stress on spatial learning and memory in the open-field tower maze across the female estrous cycle.

The purpose of the present investigation was to test how acute stress and levels of circulating estrogens together influence acquisition and retention of spatial learning, as well as explorative behaviors in female rats. We used the hippocampus-dependent Open-field Tower Maze (OFTM) task to assess acquisition followed by a retention test (reacquisition) that was given 48 h later. Immediately prior to acquisition, experimental rats were exposed to an acute restraint stress and were trained under bright lights. Female rats' estrous cycles were tracked throughout training and testing. Exposure to stress did not affect learning when levels of estrogens were low (i.e., during estrus and metestrus). However, acute stress exposure significantly lowered spatial acquisition of the female rats in the phases with rising levels of estrogens (i.e., during diestrus and proestrus). Furthermore, this stress-induced diminishment during acquisition was evident at the beginning of the retention without any presentation of stress. The present findings provide insight about the interactive relationship between stress and sex hormones on cognitive functions.

Impact of fornix lesions on tone-off delay vs tone-on trace eyeblink conditioning in rats.

Research has shown differences in the neural mechanisms that support trace and delay eyeblink conditioning. The present experiment furthered this investigation by examining the effect of electrolytic fornix lesions on acquisition of trace and delay eyeblink conditioning in the rat. Importantly, the conditioned stimulus (CS) for trace conditioning was a standard tone-on cue, and the CS for delay conditioning was either a tone-off or tone-on CS. The results showed that fornix lesions impaired trace-, but not delay conditioning in rats trained with the tone-on CS or tone-off CS. The findings are consistent with previous studies that found trace-, but not delay eyeblink conditioning is a hippocampal dependent form of associative learning. Our results also indicate that the neural pathways for tone-off delay conditioning and tone-on trace conditioning are different, even though the structural composition of a tone-off CS and the trace conditioning interval are the same cue (i.e., the absence of sound). These findings indicate that the absence of a sensory cue (i.e., tone-off CS) and the presence of a sensory cue (i.e., tone-on CS) have equivalent associative value and effectiveness for engaging the neural pathways that support delay eyeblink conditioning.

Fornix lesions impair place-, but not response-learning in the open-field tower maze.

The purpose of the present study was to examine hippocampal function for spatial learning in a land-based circular maze (i.e., the open-field tower maze [OFTM]). The OFTM, a task designed to be non-stressful, has been previously used to demonstrate the influence of gonadal hormones on spatial learning. Thus, determination of brain function for navigating in the OFTM provides an important extension to previous knowledge. Fornix lesions were used in the present experiment to disrupt hippocampal processing. After initial pre-training, rats received either an electrolytic fornix lesion surgery or a sham surgery. The rats from each surgical group were given either place- or response-training in the OFTM. The results showed that (1) lesioned place-learners required more trials than sham place-learners to solve the OFTM and (2) lesioned response-learners solved the OFTM at the same rate as sham response-learners. Our findings support the hypothesis that the hippocampus is necessary for place-, but not response-learning in the OFTM task. The OFTM is an appetitive task that does not depend on a choice between restricted directions that a rat would be required to make in a T-maze or a radial arm-maze, and does not include aversive components inherent to a Morris Water Maze or Barnes Maze. Thus, the OFTM can be used to investigate the manipulations of hippocampus-dependent spatial learning without confounding variables related to an animal's stress level.

Sex differences and the role of acute stress in the open-field tower maze.

Many studies provide evidence that differences in spatial learning exist between males and females. However, it is necessary to consider non-mnemonic factors that may influence these findings. The present experiment investigated acquisition, retention, and the effects of stress on response- and place-learning in male and female rats. Rats were trained in an open-field tower maze. Procedures were used to minimize stress in the rats, and their ability to solve place- or response-learning in the maze was determined by analyzing a response variable (i.e., first choice correct response) that was not influenced by general locomotor activity. The results revealed that male and female rats acquire place- and response-learning at the same rate even though females moved significantly faster in the maze. However, females showed better retrieval of place-, but not response-learning compared to male rats. This effect appeared to be enhanced when the rats were tested immediately following an acute restraint stress. Furthermore, both female and male rats that were exposed to acute restraint stress showed less impairment than controls when subsequently tested in a novel situation. These findings have clinical implications that a mild physiological stress response can make one more cognitively resistant to adversities later in life.

Auditory cue absence as a conditioned stimulus for delay eyeblink conditioning.

The present experiment was designed to determine if the absence of an auditory cue (i.e., a "tone-off" cue) would be an effective conditioned stimulus (CS) for delay eyeblink conditioning and to test if the medial geniculate nucleus (MGN) is part of the sensory pathway for tone-off conditioning. Rats were given paired or unpaired delay eyeblink conditioning to examine if responding to a tone-off CS was due to an associative process. An inactivation technique was performed on a separate group of rats to determine if the MGN is needed to express tone-off conditioning. The results showed that rats given paired conditioning acquired robust conditioned eyeblink responses (CRs) compared with rats given unpaired conditioning and that expression of tone-off elicited CRs was impaired when the MGN was inactivated. The findings suggest that tone-on and tone-off eyeblink conditioning may share a common neural pathway. (PsycINFO Database Record

The Miracle Fruit: An Undergraduate Laboratory Exercise in Taste Sensation and Perception.

"Miracle Fruit" is a taste-altering berry that causes sour foods to be perceived as sweet. The present paper describes a laboratory exercise that uses Miracle Fruit to educate students about the sensation and perception of taste. This laboratory exercise reinforces course material pertaining to the function of sweet taste receptors covered in a Sensation and Perception course at Christopher Newport University. Here we provide a step-by-step explanation of the methodology, and an example of data collected and analyzed by one group of students who participated in this laboratory exercise. The origins of the Miracle Fruit, the structure and the physiological function of miraculin (the glycoprotein responsible for the taste-modifying effect found in the pulp of the Miracle Fruit) were discussed before the laboratory exercise. Students then sampled foods known to target different types of tastes (i.e., sweet, sour, bitter and salty) and rated their perception of taste intensity for each food item. Next, students each consumed Miracle Fruit berries, then resampled each original food item and again recorded their perception of taste intensity ratings for these foods. The data confirmed that the sour food items were perceived sweeter after the Miracle Fruit was consumed. The students also completed a written assignment to assess what they learned about the origins, structure, and physiological function of Miracle Fruit. This hands-on laboratory exercise received positive feedback from students. The exercise can be used by other neuroscience educators to teach concepts related to the sensory system of taste.

17ß-estradiol replacement in ovariectomized female rats slows set 1 dorsolateral striatial-dependent learning and enhances learning of set 2 in an extradimensional set-shifting paradigm.

The role of estrogen in extradimensional set-shifting was evaluated with replacement of 17ß-estradiol (E2) in ovariectomized (OVX) female rats. Rats were reinforced with food when they entered an arm of a plus-maze that was distinguished by visual and/or tactile cues (Set 1). In Set 2, reinforcement was shifted to construct a new association between food and visual/tactile cues that were different from Set 1. The purpose of using this extradimensional set-shifting task was to differentiate the effect of acute or continuous E2 on the dorsolateral (DLS) versus dorsomedial (DMS) striatum and medial prefrontal cortex (mPFC), because Set 1 and 2 learning, respectively, are associated with these particular brain regions. Results showed that compared to controls, acute E2-replaced female rats required more training trials to reach criterion in Set 1. Moreover, E2-replaced females showed a significant delay in the rate of acquisition of Set 1 learning compared to controls. In Set 2 there were no group differences in perseverative errors, which are reduced by mPFC activation, or when learning took place in a previously reinforced arm, a DMS-mediated effect. Despite this, control females required more training trials to learn Set 2 compared to Set 1, suggesting that prior learning in Set 1 interfered with Set 2 performance in non-E-replaced rats. In contrast, E2 groups learned Set 2 in fewer training trials than Set 1. These data suggest that E2 facilitates set shifting, apart from any apparent enhancement of DMS or mPFC function, perhaps by interfering with DLS-mediated Set 1 learning.

Place and Response Learning in the Open-field Tower Maze.

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how "stress" might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function(1). The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats(2). Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

Effects of continuous vs. cycling estrogen replacement on the acquisition, retention and expression of place- and response-learning in the open-field tower maze.

Estrogen has been shown to either enhance or impair learning and memory in female rats. The use of different experimental paradigms or estrogen treatment regimens may contribute to these disparate findings. In order to assess the effect of different estradiol (E2) treatments on several aspects of cognition, we trained ovariectomized female rats with either continuous, cycling, or vehicle E2 replacement, in an open-field tower maze task (OFTM) designed to test reference memory in a low-stress environment. In addition, in order to compare two distinct learning and memory systems, rats were trained to use either a dorsolateral striatum-based response type learning or a hippocampal-based place type learning to solve the maze. Results showed that cyclic, but not continuous, E2 replacement facilitated the acquisition of spatial memory in place-learners. Neither E2 regimen affected acquisition in response-learners. Additionally, when all experimental groups were performing at asymptote, rats were evaluated for performance stability by changing the location of their start position in the OFTM. Both regimens of E2 disrupted the expression of spatial memory in place-learners following the novel start position. However, E2 replacement protected ovariectomized female rats from the disruption of memory expression following a start position change in response-learners. Additionally all experimental groups performed equally well when tested following a 21-day period during which rats were absent from the maze. These results suggest that E2 fluctuation is particularly important in the acquisition of hippocampal-mediated spatial learning, and that hippocampal-based memory may be subject to disruption following environmental change, while striatum-based memory is subject to protection.

Estrogen enhances the retention of spatial reference memory in the open field tower task, but disrupts the expression of spatial memory following a novel start position.

Estrogen's (E) involvement in cognition has been difficult to characterize; numerous studies show that E can both enhance and impair learning and memory. One difficulty may be that experimental paradigms often examine only a single aspect of E's involvement in cognition, for example, the role E plays in the expression of memory after learning has taken place. In addition, the effect of aversive and/or stressful features inherent to many cognitive tests may contribute to the contradictory findings. The present experiment aims to examine the effect of estradiol (E2) on several elements of cognition in a specific experimental setting. We investigated the within-subject effects of long-term E2 replacement in ovariectomized (OVX) female rats on the acquisition and retention of a hippocampal-mediated spatial reference memory task in a familiar non-threatening environment. Results show that E2-replaced rats and OVX sham-replaced rats acquired the ability to navigate an open-field tower maze in order to obtain a food reward at the same rate. Subsequent to acquisition, both E2-replaced and OVX rats performed the task at comparable levels. However, following a 21-day retention interval, non-replaced rats exhibited a significant impairment in spatial memory when returned to the maze environment, while E2-replaced rats exhibited no change in maze performance. When the OVX group was performing once again at asymptote, test trials were administered during which the rats were placed in a non-experienced start location within the maze. This novel condition significantly reduced correct responses in E2-replaced females whereas OVX controls remained unaffected. These results suggest that while the presence of E2 is not important for acquisition of spatial memory in a safe familiar environment, it improves retention of spatial memory. Data further suggests that E2 disrupts the expression of spatial reference memory following an alteration of the test conditions sustaining a habitual response, possibly by the induced emotionally-arousing state of stress.

Pharmacologic rescue of motivational deficit in an animal model of the negative symptoms of schizophrenia.

BACKGROUND: Deficits in incentive motivation, the energizing of behavior in pursuit of a goal, occur in many psychiatric disorders including schizophrenia. We previously reported deficits in both cognition and incentive motivation in a transgenic mouse model of increased striatal-specific dopamine D2 receptor (D2R) density (D2R-OE mice). This molecular alteration is observed in patients with schizophrenia, making D2R-OE mice a suitable system to study the cellular and molecular mechanisms of motivation and avolition, as well as a tool for testing potential therapies against motivational deficits. METHODS: Behavioral studies using operant conditioning methods were performed both to further characterize the incentive motivation deficit in D2R-OE mice and test a novel pharmacological treatment target that arose from an unbiased expression study performed using gene chips and was validated by quantitative reverse transcription polymerase chain reaction, in situ hybridization, and immunohistochemistry. RESULTS: The reluctance of D2R-OE mice to work is due neither to intolerance for low rates of reward, decreased reactivity to reward, nor increased sensitivity to satiety or fatigue but to a difference in willingness to work for reward. As in patients with schizophrenia, this deficit was not ameliorated by D2R blockade, suggesting that reversal of the motivational deficit by switching off the transgene results from molecular changes downstream of D2R overexpression. We observed a reversible increase in serotonin subtype 2C (5-HT2C) receptor expression in D2R-OE mice. Systemic injection of a 5-HT2C receptor antagonist increased incentive motivation in D2R-OE and control mice. CONCLUSIONS: We propose that targeting 5-HT2C receptors may be a useful approach to modulate incentive motivation in psychiatric illness.

Impaired timing precision produced by striatal D2 receptor overexpression is mediated by cognitive and motivational deficits.

Increased striatal dopamine D2 receptor activity is thought to contribute to the pathophysiology of schizophrenia. To model this condition in mice, Kellendonk et al. (2006) generated transgenic mice that selectively overexpress the D2 receptor in striatum (D2OE). Drew et al. (2007) reported that D2OE mice display deficits in interval timing and motivation. The present study further explored the impaired timing in D2OE mice. Experiment 1 assessed the role of motivation in producing timing deficits in the peak procedure and found that performance in D2OE mice was improved by increasing motivation. In addition, performance was impaired in control mice when motivation was decreased. In Experiment 2, we found that D2OE mice have no timing impairment when tested using the bisection task, a procedure in which the measure of timing performance is less influenced by motivation to respond. In Experiment 3, we also used the bisection task and found selective impairment in timing of long durations in D2OE mice. These results suggest that striatal D2 overexpression impairs timing by decreasing motivation and through its impact on working memory and/or sustained attention.

Constraints on Enhanced Extinction Resulting from Extinction Treatment in the Presence of an Added Excitor.

Three Pavlovian fear conditioning experiments with rats as subjects explored the effect of extinction in the presence of a concurrent excitor. Our aim was to explore this particular treatment, documented in previous studies to deepen extinction, with novel control groups to shed light on the processes involved in extinction. Relative to subjects extinguished on the target CS alone, Experiments 1 and 2 found across a range of parameters that any appreciable effect of facilitated extinction due to the concurrent excitor was submerged by generalization decrement going from extinction to testing. In Experiment 3 we used different durations for the target and concurrent stimuli in order to discourage configuring and an ABC renewal design to increase sensitivity, and observed diminished renewal resulting from extinction in the presence of a second excitor. Our findings suggest that there are distinct limits to the observation of extinction in the presence of an excitor and identifies some of the sources of these limitations.

Transient overexpression of striatal D2 receptors impairs operant motivation and interval timing.

The striatum receives prominent dopaminergic innervation that is integral to appetitive learning, performance, and motivation. Signaling through the dopamine D2 receptor is critical for all of these processes. For instance, drugs with high affinity for the D2 receptor potently alter timing of operant responses and modulate motivation. Recently, in an attempt to model a genetic abnormality encountered in schizophrenia, mice were generated that reversibly overexpress D2 receptors specifically in the striatum (Kellendonk et al., 2006). These mice have impairments in working memory and behavioral flexibility, components of the cognitive symptoms of schizophrenia, that are not rescued when D2 overexpression is reversed in the adult. Here we report that overexpression of striatal D2 receptors also profoundly affects operant performance, a potential index of negative symptoms. Mice overexpressing D2 exhibited impairments in the ability to time food rewards in an operant interval timing task and reduced motivation to lever press for food reward in both the operant timing task and a progressive ratio schedule of reinforcement. The motivational deficit, but not the timing deficit, was rescued in adult mice by reversing D2 overexpression with doxycycline. These results suggest that early D2 overexpression alters the organization of interval timing circuits and confirms that striatal D2 signaling in the adult regulates motivational process. Moreover, overexpression of D2 under pathological conditions such as schizophrenia and Parkinson's disease could give rise to motivational and timing deficits.

Recency-to-primacy shift in cue competition.

Three experiments were conducted using a lick-suppression preparation with rats to determine whether temporal and physical context shifts modulate the effectiveness of 2 sequentially trained blocking stimuli. Experiment 1 ascertained that it is possible to obtain blocking by conditioning rats to react to a target cue using 2 different blocking cues, each trained with a single-phase blocking paradigm. Experiment 2 showed that the more recently trained blocking stimulus was more effective (i.e., showed a recency effect) when testing was conducted immediately after training, but a long retention interval attenuated blocking by the most recently trained blocking stimulus and increased blocking by the initially trained blocking stimulus (i.e., a recency-to-primacy shift). This shift from recency to primacy affected in Experiment 2 by varying the retention interval was replicated in Experiment 3 by changing the physical context between training and testing. Taken together, the results indicate that the effectiveness of sequentially trained competing stimuli follows the same recency-to-primacy shift rule that is seen in traditional interference phenomena.