Using Postmeal Measures and Manipulations to Investigate Hippocampal Mnemonic Control of Eating Behavior.

Episodic meal-related memories provide the brain with a powerful mechanism for tracking and controlling eating behavior because they contain a detailed record of recent energy intake that likely outlasts the physiological signals generated by feeding bouts. This review briefly summarizes evidence from human participants showing that episodic meal-related memory limits later eating behavior and then describes our research aimed at investigating whether hippocampal neurons mediate the inhibitory effects of meal-related memory on subsequent feeding. Our approach has been inspired by pioneering work conducted by Ivan Izquierdo and others who used posttraining manipulations to investigate memory consolidation. This review describes the rationale and value of posttraining manipulations, how Izquierdo used them to demonstrate that dorsal hippocampal (dHC) neurons are critical for memory consolidation, and how we have adapted this strategy to investigate whether dHC neurons are necessary for mnemonic control of energy intake. I describe our evidence showing that ingestion activates the molecular processes necessary for synaptic plasticity and memory during the early postprandial period, when the memory of the meal would be undergoing consolidation, and then summarize our findings showing that neural activity in dHC neurons is critical during the early postprandial period for limiting future intake. Collectively, our evidence supports the hypothesis that dHC neurons mediate the inhibitory effects of ingestion-related memory on future intake and demonstrates that post-experience memory modulation is not confined to artificial laboratory memory tasks.

Postmeal optogenetic inhibition of dorsal hippocampal principal neurons increases future intake in a time-dependent manner.

Research involving human participants indicates that memories of recently eaten meals limit how much is eaten during subsequent eating episodes; yet, the brain regions that mediate the inhibitory effects of ingestion-related memory on future intake are largely unknown. We hypothesize that dorsal hippocampal (dHC) neurons, which are critical for episodic memories of personal experiences, mediate the inhibitory effects of ingestion-related memory on future intake. Our research program aimed at testing this hypothesis has been influenced in large part by our mentor James McGaugh and his research on posttraining manipulations. In the present study, we used an activity-guided optogenetic approach to test the prediction that if dHC glutamatergic neurons limit future intake through a process that requires memory consolidation, then inhibition should increase subsequent intake when given soon after the end of a meal but delayed inhibition should have no effect. Viral vectors containing CaMKIIα-eArchT3.0-eYFP and fiber optic probes were placed in the dHC of male Sprague-Dawley rats. Compared to intake on a day when no inhibition was given, postmeal inhibition of dHC glutamatergic neurons given for 10 min after the end of a saccharin meal increased the likelihood that rats would consume a second meal 90 min later and significantly increased the amount of saccharin solution consumed during that next meal when the neurons were no longer inhibited. Importantly, delayed inhibition given 80 min after the end of the saccharin meal did not affect subsequent intake of saccharin. Given that saccharin has minimal postingestive gastric consequences, these effects are not likely due to the timing of interoceptive visceral cues generated by the meal. These data show that dHC glutamatergic neural activity is necessary during the early postprandial period for limiting future intake and suggest that these neurons inhibit future intake by consolidating the memory of the preceding meal.

Predicting the effects of a high-energy diet on fatty liver and hippocampal-dependent memory in male rats.

OBJECTIVE: In rodents, diets exceeding nutritional requirements (i.e., high-energy diets; HED) impair hippocampal-dependent memory. Our research suggests that the effects likely involve HED-induced increases in liver lipids. In this experiment, rats were provided with diet choices to test whether voluntary consumption of a HED impairs spatial memory, whether differences in initial weight gain predict memory deficits, and whether increases in liver lipids are associated with the memory deficits. DESIGN AND METHODS: Adult male Sprague-Dawley rats were given a control diet or cafeteria-style HED for 8 weeks. Weight gain during the first 5 days on the diet was used to divide rats into a HED-Lean group and a HED-Obese group. Spatial water maze memory was tested 8 weeks later and postmortem liver lipid concentrations were quantified. RESULTS: Compared with the HED-Lean and control rats, the HED-Obese rats had impaired spatial memory and met the human diagnostic criterion of non-alcoholic fatty liver disease (>5% liver lipids relative to liver weight). Moreover, liver lipids were correlated with memory deficits. CONCLUSIONS: These findings show that voluntary consumption of a HED impairs memory, that initial weight gain predicts fatty liver and memory deficits, and that fatty liver may contribute to the memory-impairing effects of obesity.

Non-alcoholic fatty liver disease impairs hippocampal-dependent memory in male rats.

Non-alcoholic fatty liver disease (NAFLD) is a disorder observed in children and adults characterized by an accumulation of liver fat (>5% wet weight) in the absence of excessive alcohol intake. NAFLD affects 10 to 30% of the American population and is the most common cause of liver disease in the United States. NAFLD leads to serious disturbances in cardiovascular and hormonal function; however, possible effects on brain function have been overlooked. The aims of the present study were to test whether diet-induced NAFLD impairs hippocampal-dependent memory and to determine whether any observed deficits are associated with changes in hippocampal insulin signaling or concentrations of brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-1 (IGF-1). Post-weanling male Sprague-Dawley rats were fed a high fructose (60% of calories) or control diet for 12 weeks and then trained and tested in a spatial water maze. NAFLD was confirmed with postmortem measures of liver mass and liver lipid concentrations. NAFLD did not affect acquisition of the spatial water maze, but did impair retention tested 48 h later. Specifically, both groups demonstrated similar decreases in latency to swim to the escape platform over training trials, but on the memory test NAFLD rats took longer to reach the platform and made fewer visits to the platform location than control diet rats. There were no differences between the groups in terms of insulin-stimulated phosphorylation of insulin receptor ß subunit (IR-ß) and protein kinase B (PKB/AKT) in hippocampal slices or hippocampal BDNF or IGF-1 concentrations. Thus, these data indicate that NAFLD impairs hippocampal-dependent memory function and that the deficit does not appear attributable to alterations in hippocampal insulin signaling or hippocampal BDNF or IGF-1 concentrations.

A high fructose diet impairs spatial memory in male rats.

Over the past three decades there has been a substantial increase in the amount of fructose consumed by North Americans. Recent evidence from rodents indicates that hippocampal insulin signaling facilitates memory and excessive fructose consumption produces hippocampal insulin resistance. Based on this evidence, the present study tested the hypothesis that a high fructose diet would impair hippocampal-dependent memory. Adult male Sprague-Dawley rats (postnatal day 61) were fed either a control (0% fructose) or high fructose diet (60% of calories). Food intake and body mass were measured regularly. After 19 weeks, the rats were given 3 days of training (8 trials/day) in a spatial version of the water maze task, and retention performance was probed 48 h later. The high fructose diet did not affect acquisition of the task, but did impair performance on the retention test. Specifically, rats fed a high fructose diet displayed significantly longer latencies to reach the area where the platform had been located, made significantly fewer approaches to that area, and spent significantly less time in the target quadrant than did control diet rats. There was no difference in swim speed between the two groups. The retention deficits correlated significantly with fructose-induced elevations of plasma triglyceride concentrations. Consequently, the impaired spatial water maze retention performance seen with the high fructose diet may have been attributable, at least in part, to fructose-induced increases in plasma triglycerides.

Infusions of physostigmine into the hippocampus or the entorhinal cortex attenuate avoidance retention deficits produced by intra-septal infusions of the GABA agonist muscimol.

Septal gamma-aminobutyric acid (GABA) receptor activation is known to disrupt memory formation, although the mechanisms underlying this impairment remain unclear. The present study explored the possibility that high levels of septal GABA receptor activity might impair memory by down-regulating acetylcholine (ACh) function in archicortex and entorhinal cortex. To test this possibility, rats were trained on an avoidance task 15 min after receiving intra-septal infusions of vehicle or muscimol (5 nmol/0.5 microl) combined with unilateral intra-hippocampal (10 microl/1 microl) or intra-entorhinal cortex (1.875 microg/0.25 microl) infusions of vehicle or the acetylcholinesterase inhibitor physostigmine. We demonstrate that these infusions do not alter acquisition performance on a continuous multiple trial inhibitory avoidance task. However, intra-septal infusions of muscimol dramatically impair retention performance 48 h later. More importantly, infusions of physostigmine into the hippocampus or the entorhinal cortex, at doses that do not influence acquisition or retention performance when infused alone, attenuate the impairing effects of the muscimol infusions on retention. We suggest that high levels of septal GABA receptor activity might impair memory by down-regulating ACh levels in the hippocampal region, and that such memory impairments can be ameliorated by increasing ACh levels in the hippocampus or entorhinal cortex.

Emotionally arousing pictures increase blood glucose levels and enhance recall.

Arousal enhances memory in human participants and this enhancing effect is likely due to the release of peripheral epinephrine. As epinephrine does not readily enter the brain, one way that peripheral epinephrine may enhance memory is by increasing circulating blood glucose levels. The present study investigated the possibility that emotionally arousing color pictures would improve memory and elevate blood glucose levels in human participants. Blood glucose levels were measured before, 15 min, and 30 min after male university students viewed 60 emotionally arousing or relatively neutral pictures. Participants viewed each picture for 6 s and then had 10 s to rate the arousal (emotional intensity) and valence (pleasantness) of each picture. A free-recall memory test was given 30 min after the last picture was viewed. Although the emotionally arousing and neutral picture sets were given comparable valence ratings, participants who viewed the emotionally arousing pictures rated the pictures as being more arousing, recalled more pictures, and had higher blood glucose levels after viewing the pictures than did participants who viewed the neutral pictures. These findings indicate that emotionally arousing pictures increase blood glucose levels and enhance memory, and that this effect is not due to differences in the degree of pleasantness of the stimuli. These findings support the possibility that increases in circulating blood glucose levels in response to emotional arousal may be part of the biological mechanism that allows emotional arousal to enhance memory.

Effects of the antidepressant/antipanic drug phenelzine on alanine and alanine transaminase in rat brain.

1. Phenelzine (PLZ) is an antidepressant with anxiolytic properties. Acute and chronic PLZ administration increase brain GABA levels, an effect due, at least in part, to an inhibition of the activity of the GABA metabolizing enzyme, GABA transaminase (GABA-T). 2. Previous preliminary reports have indicated that acute PLZ treatment also elevates brain alanine levels. As with GABA, the metabolism of alanine involves a pyridoxal phosphate-dependent transaminase. 3. In the study reported here, the effects of acute PLZ treatment on the levels of various amino acids, some of which are also metabolized by pyridoxal phosphate-dependent transaminases were compared in rat whole brain. Of the 6 amino acids investigated, only GABA and alanine levels were elevated (in a time- and dose-dependent manner). 4. The elevation in brain alanine levels could be explained, at least in part, by a time- and dose-dependent inhibitory effect of PLZ on alanine transaminase (ALA-T), although as with GABA the increases are higher than expected from the degree of enzyme inhibition produced. In addition, we also showed that the elevation in alanine levels and the inhibition of alanine transaminase in the brain are retained after 14 days of PLZ treatment, and that PLZ produces a marked increase in extracellular levels of alanine. 5. These results are discussed in terms of their relevance to synaptic function and to the pharmacological profile of PLZ.

Increasing acetylcholine levels in the hippocampus or entorhinal cortex reverses the impairing effects of septal GABA receptor activation on spontaneous alternation.

Intra-septal infusions of the gamma-aminobutyric acid (GABA) agonist muscimol impair learning and memory in a variety of tasks. This experiment determined whether hippocampal or entorhinal infusions of the acetylcholinesterase inhibitor physostigmine would reverse such impairing effects on spontaneous alternation performance, a measure of spatial working memory. Male Sprague-Dawley rats were given intra-septal infusions of vehicle or muscimol (1 nmole/0.5 microL) combined with unilateral intra-hippocampal or intra-entorhinal infusions of vehicle or physostigmine (10 microg/microL for the hippocampus; 7.5 microg/microL or 1.875 microg/0.25 microL for the entorhinal cortex). Fifteen minutes later, spontaneous alternation performance was assessed. The results indicated that intra-septal infusions of muscimol significantly decreased percentage-of-alternation scores, whereas intra-hippocampal or intra-entorhinal infusions of physostigmine had no effect. More importantly, intra-hippocampal or intra-entorhinal infusions of physostigmine, at doses that did not influence performance when administered alone, completely reversed the impairing effects of the muscimol infusions. These findings indicate that increasing cholinergic levels in the hippocampus or entorhinal cortex is sufficient to reverse the impairing effects of septal GABA receptor activation and support the hypothesis that the impairing effects of septal GABAergic activity involve cholinergic processes in the hippocampus and the entorhinal cortex.

Amygdala lesions do not impair shock-probe avoidance retention performance.

The present experiment used the shock-probe paradigm, a procedure usually used to assess anxiolytic processes, to assess memory in amygdala-lesioned rats. Rats were placed in a chamber that contained a probe protruding from 1 of 4 walls and were kept there for 15 min after they contacted the probe. For half the rats, the probe was electrified (2 mA). Four days later, sham or neurotoxic amygdala lesions were induced. Retention performance was assessed 8 days later by measuring the latency to contact the probe and the number of contact-induced shocks. The results indicated that, although shock-naive amygdala-lesioned rats were impaired on the 2nd shock-probe test, shock-experienced amygdala-lesioned rats were not. These data indicate that the memory of a shock experience, as indexed with a shock-probe avoidance response, is spared in rats with large amygdala lesions.

Time-dependent changes in brain monoamine oxidase activity and in brain levels of monoamines and amino acids following acute administration of the antidepressant/antipanic drug phenelzine.

Phenelzine (PLZ) is a non-selective monoamine oxidase (MAO) inhibitor commonly used to treat depression and panic disorder. Acute administration of PLZ produces several neurochemical changes, including an increase in brain levels of the catecholamines norepinephrine (NE) and dopamine (DA), of 5-hydroxytryptamine (5-HT), and of the amino acids alanine and gamma-aminobutyric acid (GABA). The goal of the present series of experiments was to characterize the time course of these PLZ-induced changes. Male Sprague-Dawley rats were sacrificed 6, 24, 48, 96, 168, or 336 hr after acute PLZ administration (15 or 30 mg/kg, i.p., based on free base weight). Whole brain levels of monoamines and amino acids were determined using HPLC, and MAO A and B activities were determined using a radiochemical procedure. The results indicated that PLZ changed amino acid levels 6 and 24 hr after injection, but not 48 hr later. In contrast, the effects of PLZ on MAO activity and monoamines were longer-lasting. For example, PLZ-induced increases in dopamine and 5-HT were observed 1 week after injection, and PLZ-induced inhibition of MAO activity persisted for 2 weeks. Thus, in addition to demonstrating that the effects of PLZ on MAO activity and monoamines were long-lasting, these results indicate that the effects of PLZ on MAO activity and on brain levels of monoamines and amino acids are temporally dissociated. These findings regarding the long-term effects of PLZ on neurochemistry will have considerable critical implications for the design and interpretation of behavioral studies of the acute effects of PLZ.

Task-dependent effects of the antidepressant/antipanic drug phenelzine on memory.

Phenelzine (PLZ) is a non-selective monoamine oxidase (MAO) inhibitor commonly used to treat depression and panic disorder. In addition to increasing levels of biogenic amines in the brain, PLZ elevates brain levels of the amino acid gamma-aminobutyric acid (GABA; Baker et al. 1991; present study). Given the extensive evidence implicating biogenic amines and GABA in mnemonic processes, PLZ may affect learning and memory. To investigate this possibility, male Sprague-Dawley rats were given PLZ sulfate (15 or 30 mg/kg, based on free base weight) 2 h prior to training in a continuous multiple trial inhibitory avoidance (CMIA) and spatial water maze task. Retention was assessed 48 h later. The results indicated that PLZ enhanced CMIA and impaired water maze retention performance. Compared to control rats, rats given PLZ took significantly longer to re-enter the shock compartment and swam longer distances before reaching the escape platform on the retention tests. These effects of PLZ did not appear to be the result of PLZ-induced changes in acquisition or retrieval processes, activity levels, or footshock sensitivity. Combined, these findings indicate that PLZ influences memory in a task-dependent manner. These differential effects of PLZ may be the result of contrasting influences of GABA and biogenic amines on memory.

Intraseptal infusions of muscimol impair spontaneous alternation performance: infusions of glucose into the hippocampus, but not the medial septum, reverse the deficit.

As observed with intraseptal injections of opioid receptor agonists, direct infusions of GABAergic receptor agonists into the medial septum impair performance on several tasks that involve spatial or working memory processes in rats. Because the effects of opioid-induced impairments can be reliably reversed by concomitant intraseptal infusions of glucose, the experiments reported here determined whether impairments produced by GABAergic agonists would similarly be reversed by glucose. The findings of Experiment 1 showed, in male Sprague-Dawley rats, that intraseptal infusions of the GABA agonist muscimol (1 or 3 nmol/0.5 microliter) impaired spontaneous alternation performance. The results of Experiment 2 indicated that intraseptal infusions of glucose (8, 17, or 33 nmol) or glutamate (15 or 30 nmol) did not attenuate the muscimol-induced deficit on spontaneous alternation performance, whereas infusions of the GABAergic antagonist bicuculline methiodide (0.1 nmol) did. However, the findings of Experiment 3 indicated that glucose injections (50 nmol/0.5 microliter) into the hippocampus did reverse the impairing effect of the intraseptal muscimol infusions. Combined, these findings suggest that the neurochemical regulation of learning and memory may involve hierarchical interactions between particular neurotransmitter and neuroanatomical systems. Specifically, medial septal GABAergic effects on spontaneous alternation prevail over those of glucose or glutamate in the medial septum, but are overridden by the effects of glucose in the hippocampus.

Intra-septal infusions of glucose potentiate inhibitory avoidance deficits when co-infused with the GABA agonist muscimol.

This experiment examined the effects of co-infusions of glucose with the gamma-aminobutyric acid (GABA) agonist muscimol into the medial septum on memory for inhibitory avoidance learning. Co-infusions of muscimol (3 nmol) and glucose (33 nmol) impaired memory, but neither drug did so when administered alone. Thus, although glucose typically reverses memory deficits, these results indicate that glucose potentiates the memory-impairing effects of a GABA agonist.

Ibotenic acid lesions of the amygdala basolateral complex or central nucleus differentially effect the response to reductions in reward.

The present study examined the role of the amygdala in the acquisition and expression of the Crespi effect (Crespi, L.P., Quantitative variation in incentive and performance in the white rat, Am. J. Psychol., 55 (1942) 467-517), also known as successive negative behavioral contrast. In Experiment One rats with bilateral amygdala cannulae were trained to run a straight alley for either a large (ten pellet) or small (one pellet) food reward. After 8 days of training, half of the rats in each reward condition received vehicle or ibotenic acid administered bilaterally into the amygdala. After 4 days of recovery from the induction of the lesions, training resumed. On Day 12 of training, the reward for rats in the large reward condition was shifted to one pellet and this reward level was maintained for the next 4 days of training. Both the lesioned and unlesioned shifted rats exhibited increased latencies to the reduction. However, shifted lesioned rats displayed a more persistent increase in latencies than shifted unlesioned rats, exhibiting significantly longer latencies than those of unlesioned rats by Day 15. This finding suggests that large amygdala lesions may impair learning of the appetitive value of the small reward. Experiment Two examined the effects of discrete ibotenic acid lesions of either the central nucleus or basolateral/lateral complex of the amygdala. Lesions of the central nucleus produced results similar to those of Experiment One. However, in Experiment Two the performance of shifted unlesioned and lesioned groups diverged significantly 1 day earlier, on Day 14. In contrast, lesions of the basolateral/lateral complex reduced the duration of the contrast effect. Shifted lesioned rats exhibited significantly lower latencies than shifted unlesioned rats by the first postshift day, Day 13. This finding suggests that the basolateral/lateral complex may be involved in learning about, or expressing the response to, the aversiveness of reward reduction.

Spared retention of inhibitory avoidance learning after posttraining amygdala lesions.

Previous findings indicate that the memory-impairing effects of posttraining amygdala lesions are attenuated by increasing the number of training trials given prior to the induction of the lesion. The aim of this experiment was to determine whether the degree of impairment is also influenced by the footshock intensity used during training. Rats were given 1 trial of inhibitory avoidance (IA) training with either no footshock or a footshock at 1 of 3 intensities. Sham or neurotoxic amygdala lesions were induced 1 week later. On a retention test performed 4 days after surgery, the performance of all amygdala-lesioned rats given footshock training, including those given the lowest training footshock, was better than that of amygdala-lesioned rats given no training footshock. These findings of preserved retention of IA learning in rats given posttraining amygdala lesions do not support a general hypothesis that the amygdala is a locus of permanent changes underlying aversively motivated learning.

Footshock facilitates the expression of aversively motivated memory in rats given post-training amygdala basolateral complex lesions.

We previously reported that increased training in an escape task partially attenuates the memory impairment produced by large amygdala lesions induced 1 week following training. The present study examined the effect of amount of preoperative training on the retention of rats with lesions restricted to the amygdala basolateral complex. Rats received 1 or 10 training trials in a footshock-motivated escape task and 1 week later sham lesions or neurotoxic lesions of the basolateral complex. Four days after recovery from the surgery they were tested for inhibitory avoidance retention and then 2 days later given continuous multiple trial inhibitory avoidance training (CMIA) in the same apparatus. The basolateral complex lesions significantly decreased the retention latencies of rats given 1 or 10 trials. However, following administration of footshock on the CMIA task, the performance of the lesioned rats reflected the degree of preoperative escape training. The basolateral complex lesions also increased open field locomotor activity, an effect that may have contributed to the shorter retention latencies in lesioned animals. These findings indicate that an intact amygdala basolateral complex is not critical for the retention of the escape training.

Memory of rats with amygdala lesions induced 30 days after footshock-motivated escape training reflects degree of original training.

This experiment was conducted to determine whether the amount of preoperative training influences the effects, on retention, of amygdala lesions induced 30 days after escape training. Rats received 1 or 10 footshock-motivated escape training trials; 30 days later, sham or neurotoxic amygdala lesions were induced. Results of an inhibitory avoidance test performed 4 days after surgery indicated that amygdala lesions impaired retention performance; however, increased preoperative training partially attenuated the retention deficit. Increased preoperative training also attenuated the impairing effects of the lesions on retention assessed in a continuous multiple-trial inhibitory avoidance task given 36 days after the original escape training. The finding that amygdala-lesioned rats remembered the escape training suggests that the amygdala is not a critical locus of the changes underlying the long-term retention of footshock-motivated escape training.

Posttraining infusion of lidocaine into the amygdala basolateral complex impairs retention of inhibitory avoidance training.

The present experiment examined the role of the central nucleus and basolateral complex in the retention of inhibitory avoidance training by reversibly inactivating these regions with lidocaine immediately following training. Male Sprague-Dawley rats were surgically implanted bilaterally with cannulae aimed at the central nucleus or the basolateral complex. One week later, they received one trial inhibitory avoidance training (0.45 mA; 1 s), followed immediately by infusions of lidocaine hydrochloride or buffer (10 micrograms/0.25 microliters). Retention was tested 2 days after training. Immediate posttraining infusions of lidocaine into the central nucleus did not affect retention performance; in contrast, immediate posttraining infusions of lidocaine into the basolateral complex significantly impaired retention performance. In addition, the effect of posttraining infusions of lidocaine into the basolateral complex was time-dependent: infusions administered 6 h after training also impaired memory, but infusions administered 24 h after training had no effect. Immediate posttraining infusions of lidocaine also impaired the retention performance of rats trained with a more intense footshock (0.75 mA). However, at the higher footshock intensity, administration of lidocaine 6 h after training had no effect on retention performance. The time- and footshock-dependent retrograde impairment of memory produced by posttraining reversible inactivation of the basolateral complex suggests that this region of the amygdala is involved in the consolidation of memory for inhibitory avoidance training.

Neuromodulatory systems and memory storage: role of the amygdala.

This article reviews findings of research examining the interaction of peripheral adrenergic systems with cholinergic, opioid peptidergic and GABAergic systems in modulating memory storage. It is well established that retention is enhanced by posttraining systemic or intra-amygdala injections of adrenergic agonists, opiate antagonists and GABAergic antagonists. These influences appear to be mediated by activation of NE receptors within the amygdala, as intra-amygdala injections of beta-adrenergic antagonists block the memory-modulating effects of hormones and drugs affecting these systems. Furthermore, these influences also appear to involve, at a subsequent step, activation of a cholinergic system: atropine blocks the memory-enhancing effects of adrenergic agonists and opiate and GABAergic antagonists and oxotremorine attenuate the memory-impairing effects of opiate agonists and GABAergic agonists. These findings suggest that the amygdala integrates the memory-modulating effects of neuromodulatory systems activated by learning experiences.