The influences of sex, rearing environment, and neonatal choline dietary supplementation on spatial and nonspatial learning and memory in adult rats

The potential facilitative effects of early environmental enrichment and perinatal choline chloride dietary supplementation on male and female adult rats' learning and memory were examined using a "stimulus-elicited investigative," and a social/observational learning-cued spatial memory paradigm. Male and female animals were either maintained in a standard lighted colony (SC) or were given supplementary exposure to a complex environment (EC) for 2 hr daily from 24-90 days of age. In each case, half of the animals were exposed to the choline supplementation both prenatally and postnatally for 24 days. In one paradigm, the 90-day-old EC rats were found to be significantly more responsive than SC rats to each change in the spatial relationships of objects contained in an open field. Neither sex nor early diet of the animals were much of a factor in the investigative behavior observed. In the second paradigm, the effects of the perinatal choline diet did interact with those of sex and postnatal environment to alter the impact of social/observational experience on the acquisition and memory of place in the water maze. The choline-treated EC males were the most influenced by their experience seeing a demonstrator swim to a platform location. The present study provides some further insight into the scope of the long-term functional enhancements produced by perinatal choline supplementation and EC in male and female animals and relates these effects to common modifications to targets of cholinergic basal forebrain systems.     

Problem-solving behaviour in apomorphine-susceptible and unsusceptible rats.

Pharmacogenetically selected apomorphine-susceptible (APO-SUS) and apomorphine-unsusceptible (APO-UNSUS) rats were trained in a discrimination learning paradigm. After the initial discrimination task was solved, reinforcement contingencies were reversed. No differences between APO-SUS and APO-UNSUS animals were found in the rate of learning. However, negative transfer from the initial discrimination to its reversal was less for the APO-SUS rats than for the APO-UNSUS rats. Moreover, the APO-SUS rats responded more to the relevant dimension (light) than the APO-UNSUS rats in the last 100 trials before solving the initial problem as well as the reversal. During overtraining on the first problem, APO-SUS animals responded less to an irrelevant dimension (position of the lever) than APO-UNSUS animals. In the first 100 trials of the reversal APO-SUS rats responded more to another irrelevant dimension (noise) than APO-UNSUS rats. The data show that APO-SUS and APO-UNSUS rats used the various dimensions (visual, auditory, and spatial) differently in the chosen discrimination learning paradigm. It is concluded that the interline differences found are the consequences of the interline differences in the dopaminergic activity of the ventral and dorsal striatum.     

Pre- and postnatal choline supplementation produces long-term facilitation of spatial memory

Although research has demonstrated that short-term improvement in memory function of adult rats can occur when the availability of precursors for the neurotransmitter acetylcholine is increased, little is known about whether memory function of adult rats can be permanently altered by precursor supplementation during early development. In the present study, male albino rats were exposed to choline chloride supplementation both prenatally (through the diet of pregnant rats) and postnatally (subcutaneous injections). At 60 days of age rats were tested on a 12- and 18-arm radial maze task. Results indicated that compared to control littermates, perinatal choline-treated rats showed more accurate performance on both working and reference memory components of the task. This performance difference was apparent on the first block of sessions and continued throughout training. Further analysis revealed that the difference between choline and control rats is not due to use of differential response or cue-use strategies. Instead, it appears that choline induced performance differences are due to long-term enhancement of spatial memory capacity and precision.     

Cholinergic blockade effects on spatial integration versus cue discrimination performance

A within-subjects investigation was conducted to determine the effects of central versus peripheral cholinergic blockade in animals tested either on a spatial integration task in which the possibility of rule learning was also available or on a visual discrimination task in which the daily location of food was marked by a distinctive visual stimulus pattern. All testing was conducted on the Maier three-table apparatus. It was found that the only effect of the peripheral cholinergic blockade on the performance of either task group was to produce a decrease in exploratory behavior. In contrast, central cholinergic blockade markedly impaired spatial integration performance; however, it did not impair the ability of animals in rule learning or visual discrimination learning. It was also found that central cholinergic blockade impaired the animal's tendency to enter all tables before reentering a given table during the exploratory phase of the daily session. This finding was interpreted as reflecting an impairment of working memory for spatial information, rather than a general impairment in working memory, and this interpretation was applied to the explanation of the deficit in the spatial integration performance.     

Lesions of the anterior thalamic nuclei and intralaminar thalamic nuclei: place and visual discrimination learning in the water maze

Medial thalamic damage produces memory deficits in humans (e.g., Korsakoff’s syndrome) and experimental animals. Both the anterior thalamic nuclei (ATN) and rostral intralaminar plus adjacent lateral thalamic nuclei (ILN/LT) have been implicated. Based on the differences in their main connections with other neural structures, we tested the prediction that ATN lesions would selectively impair acquisition of spatial location discrimination, reflecting a hippocampal system deficit, whereas ILN/LT lesions would impair acquisition of visual pattern discrimination, reflecting a striatal system deficit. Half the rats were first trained in a spatial task in a water maze before switching to a visual task in the same maze, while the remainder were tested with the reverse order of tasks. Compared with sham-operated controls, (1) rats with ATN lesions showed impaired place learning, but normal visual discrimination learning, (2) rats with ILN/LT lesions showed no deficit on either task. Rats with ATN lesions were also hyperactive when their home cage was placed in a novel room and remained more active than ILN/LT or SHAM rats for the subsequent 21 h, especially during the nocturnal phase. These findings confirmed the influence of ATN lesions on spatial learning, but failed to support the view that ILN/LT lesions disrupt striatal-dependent memory.     

Spatial learning and memory following fimbria-fornix transection and grafting of fetal septal neurons to the hippocampus

Summary The ability of intrahippocampal grafts of fetal septal-diagonal band tissue, rich in developing cholinergic neurons, to ameliorate cognitive impairments induced by bilateral fimbria-fornix transections in rats was examined in three experiments using the Morris water-maze to test different aspects of spatial memory. Experiment 1. Rats with fimbria-fornix lesions received either septal cell suspension grafts or solid septal grafts; normal rats and rats with lesions alone were used as controls. Sixteen weeks after surgery, the rats' spatial learning and memory were tested in the water-maze using a place test, designed to investigate place navigation performance, in which rats learned to escape from the water by swimming to a platform hidden beneath the water's surface. After 5 days of training, the rats were given a spatial probe test in which the platform was removed from the tank to test spatial reference memory. Experiment 2. The same rats used in Exp. 1 were tested in a delayed-match-to-sample, working memory version of the water-maze task. The platform was located in one of two possible locations during each trial, which was composed of 2 swims. If the rat remembered the location of the platform on the 2nd swim of a trial, it should find the platform more quickly on that swim, and thereby demonstrate working memory. Experiment 3. Prior to receiving fimbria-fornix lesions, normal rats were trained in a modification of the water-maze task using alternating cue navigation and place navigation trials (i.e., with visible or non-visible escape platforms). The retention and reacquisition of the place task and the spatial probe test were examined in repeated tests up to 6 months after the lesion and intrahippocampal grafting of septal cell suspensions. The effects of central muscarinic cholinergic receptor blockade with atropine were also tested. Normal rats performed well in both the place and spatial probe tests. In contrast, rats with fimbria-fornix lesions only were unable to acquire or retain spatial information in any test. Instead, these rats adopted a random, nonspatial search strategy, whereby their latencies to find the platform decreased in the place navigation tasks. Sixty to 80% of the rats with septal suspension or solid grafts had recovered place navigation, i.e., the ability to locate the platform site in the tank, in Exp. 1 and 3, and they showed a significantly improved performance in the working memory test in Exp. 2. Atropine abolished the recovered place navigation in the grafted rats, whereas normal rats were impaired to a lesser extent. In contrast, atropine had no effect on the non-spatial strategy adopted by rats with fimbria-fornix lesions only. The results show that: (1) fimbria-fornix lesions disrupt spatial learning and memory in both naive and pretrained rats; (2) with extended training the fimbria-fornix lesioned rats develop an efficient non-spatial strategy, which enables them to reduce their escape latency to levels close to those of intact controls; (3) intrahippocampal septal grafts can restore the ability of the lesioned rats to use spatial cues in the localization of the platform site; and (4) the behavioural recovery produced by grafts is dependent upon an atropine sensitive mechanism.     

Lack of task specificity and absence of posttraining effects of atropine on learning

Three experiments are reported whose purpose was to examine the effect of the cholinergic antagonist atropine on the acquisition of different learning tasks known to be sensitive or insensitive to impairment by hippocampal lesions; on the retention of performance acquired in the absence of the drug; and on memory consolidation immediately after daily training trials. In Experiment 1, atropine sulfate (10 or 50 mg/kg, ip), injected 30 min prior to training, severely impaired learning of both spatial and nonspatial discrimination tasks when compared with saline or atropine methylnitrate (50 mg/kg). In Experiment 2, atropine sulfate (50 mg/kg) also impaired spatial discrimination accuracy in rats previously trained to asymptote under drug-free conditions. These deficits were not due to either peripheral drug effects or gross sensorimotor impairments. In Experiment 3, daily posttraining injections of atropine sulfate (50 mg/kg) failed to influence either learning or subsequent retention of place navigation in rats that were trained to find a single hidden escape platform. The data confirm that profound learning deficits occur when training is conducted under atropine but offer no support to the hypothesis that cholinergic neurons play an important role in memory consolidation or other posttraining processes. Furthermore, these results point to dissimilarities between the behavioral impairments induced by cholinergic blockade and hippocampal lesions under appropriate test regimes.     

A comparison of discrimination and reversal learning for olfactory and visual stimuli in aged rats

The present study investigated age-related differences in discrimination and reversal learning for olfactory and visual stimuli in 6-month and 24-month-old rats. Rats were trained to discriminate between two pseudo-randomly selected odors or objects. Once each animal reached a criterion on discrimination trials, the reward contingencies were reversed. Young and aged rats acquired the olfactory and visual discrimination tasks at similar rates. However, on reversal trials, aged rats required significantly more trials to reach the learning criterion on both the olfactory and visual reversal tasks than young rats. The deficit in reversal learning was comparable for odors and objects. Furthermore, the results showed that rats acquired the olfactory task more readily than the visual task. The present study represents the first examination of age-related differences in reversal learning using the same paradigm for odors and objects to facilitate cross-modal comparisons. The results may have important implications for the selection of memory paradigms for future research studies on aging.     

Supplemental choline during the periweaning period protects against trace conditioning impairments attributable to post-training ethanol exposure in adolescent rats

Supplemental choline during early stages of development can result in long-lasting improvements to memory function. In addition, pre- or postnatal choline has been shown to be protective against some of the adverse effects of early alcohol exposure. The present experiment examined whether supplemental choline given to rats would protect against the effects of posttraining alcohol administration on trace fear conditioning. Posttraining alcohol exposure in adolescent rats results in poor performance in this hippocampus-dependent task, although delay conditioning is unaffected. Here, rats were given an s.c. injection of either saline or choline chloride daily on postnatal days (PD) 15-26. On PD 30 subjects were trained in a trace fear conditioning procedure. For the next 3 days animals were administered 2.5 g/kg ethanol or water control, and conditional stimulus (CS)-elicited freezing was measured on PD 34. Results indicated that posttraining alcohol disrupted the expression of trace conditioning and that supplemental choline on PD 15-26 was protective against this effect. That is, choline-treated animals subsequently given posttraining ethanol performed as well as animals not given ethanol. These results indicate that supplemental choline given during the periweaning period protects against ethanol-induced impairments in a hippocampus-dependent learning task. Findings contribute to the growing literature showing improvements in learning and memory in subjects given extra dietary choline during critical periods of brain development.     

Organizational changes in cholinergic activity and enhanced visuospatial memory as a function of choline administered prenatally or postnatally or both

This experiment was an examination of the effects of supplemental dietary choline chloride given prenatally (to the diet of pregnant rats) and postnatally (intubed directly into the stomachs of rat pups) on memory function and neurochemical measures of brain cholinergic activity of male albino rats when they became adults. The data demonstrate that perinatal choline supplementation causes (a) long-term facilitative effects on working and reference memory components of a 12-arm radial maze task, and (b) alternations of muscarinic receptor density as indexed by [3H]quinuclidinyl benzilate (QNB) binding and choline acetyltransferase (ChAT) levels in the hippocampus and frontal cortex of adult rats. An analysis of the relationship between these organizational changes in brain and memory function indicated that the ChAT-to-QNB ratio in the hippocampus is highly correlated with working memory errors, and this ratio in the frontal cortex is highly correlated with reference memory errors.     

Double dissociation of fornix and caudate nucleus lesions on acquisition of two water maze tasks: further evidence for multiple memory systems.

The present study examined the effect of lesions of the caudate nucleus or fimbria-fornix on the acquisition of two water maze tasks. In both tasks, two rubber balls with different visual patterns were used as platforms (i.e., cues). The "correct" cue was attached to a submerged rectangular platform and could be mounted by an animal to escape the water. The "incorrect" cue was attached to a thin round pedestal and could not be mounted. In a spatial version of the task, the correct cue was located in the same quadrant of the maze on all trials, whereas the visual pattern on the cue was varied from trial to trial. Lesions of the fornix, but not the caudate nucleus, impaired acquisition of this spatial task in relation to control animals. In a simultaneous visual discrimination version of the task, the correct cue on all trials was one with a specific visual pattern, and the spatial location of the correct cue was varied from trial to trial. Lesions of the caudate nucleus, but not the fornix, impaired acquisition of this visual discrimination task in relation to control animals. The double dissociation observed supports the hypothesis that the hippocampus and caudate nucleus are parts of systems that differ in the type of memory they mediate.     

Effects of morphine dependence on the performance of rats in reference and working versions of the water maze

Numerous studies have dealt with the role of opiate system in tasks aimed at measurement of cognitive behavior, but the role of morphine dependence on learning and memory is still controversial. In this study chronic exposure to morphine was employed to evaluate learning ability and spatial short-term memory (working memory) and long-term memory (reference memory) in the water maze task. Male albino rats were made dependent by chronic administration of morphine in drinking water that lasted at least 21 days. In Experiment 1, the performance of animals was evaluated in reference memory version of the water maze. Rats were submitted to a session of 6 trials for 6 consecutive days to find the submerged platform that was located in the center of a quadrant. Latency and traveled distance to find the platform were measured as indexes of learning. Memory retention was tested 24 h after the last training session in a probe trial (60 s) in which there was no platform and the time spent in each quadrant of the water maze was recorded. Results indicated that latency and traveled distance to find the platform were same in control and dependent rats during training days, but during the probe test morphine-dependent group spent significantly less time in the target quadrant. In Experiment 2, training on working memory version of the water maze task was started. Only two trials per day were given until the performance of animals was stabilized (at least 5 days). Final test was done at day 6. Acquisition-retention interval was 75 min. No significant differences were found on acquisition and retention trials between morphine and control groups. Our findings indicate that chronic exposure to morphine did not impair learning ability, but partially impaired retention of spatial long-term (reference) memory. Moreover, dependence on morphine did not affect either acquisition or retention of spatial short (working) memory.     

Effects of distraction and stress on delayed matching-to-place performance in aged rats

The performance of male Long-Evans rats on the delayed match-to-place (DMP) version of the Morris water maze was assessed in two separate experiments; the first compared young (4 months) with middle-aged (16 months) rats, whereas the second compared middle-aged (14 months) with old (26 months) rats. Old rats continued to use a short-term memory strategy on the DMP task, but their performance on both search and recall trials was impaired relative to that of middle-aged animals. Rats of all ages habituated rapidly to visual distraction and the performance of old rats was not affected by exposure to a mild predator stress in the form of cat urine. The performance of the middle-aged rats did not differ significantly from that of young rats, even when they were challenged on recall trials by visual distraction or by exposure to predator odour. These results do not provide strong support for the prediction that visual distraction and psychological stress would interact with age in affecting spatial short-term memory in Long-Evans rats.     

Nerve growth factor improves spatial learning and restores hippocampal cholinergic fibers in rats withdrawn from chronic treatment with ethanol

The cholinergic septohippocampal pathway has long been known to be important for learning and memory. Prolonged intake of ethanol causes enduring memory deficits, which are paralleled by partial depletion of hippocampal cholinergic afferents. We hypothesized that exogenous supply of nerve growth factor (NGF), known to serve as a trophic substance for septal cholinergic neurons, can revert the ethanol-induced changes in the septohippocampal cholinergic system. Adult rats were given a 20% ethanol solution as their only source of fluid for 6 months. During the first 4 weeks after the animals were withdrawn from ethanol, they were intraventricularly infused with either NGF or vehicle alone via implanted osmotic minipumps. The vehicle-infused withdrawn animals showed impaired performance on a spatial reference memory version of the Morris water maze task, both during the task acquisition and on the retention test. In contrast, NGF-treated withdrawn rats were able to learn the task as well as controls, and significantly outperformed the vehicle-infused withdrawn rats. The histological analysis revealed that, in the latter group, the length density of fibers immunoreactive to choline acetyltransferase was reduced relative to control values by approximately 25%, as measured in the dentate gyrus and regio superior of the hippocampal formation. However, in NGF-treated withdrawn rats, the length density of these fibers was identical to that of control rats. These data provide support to the notion that NGF is capable of ameliorating memory deficits and restoring septohippocampal cholinergic projections following chronic treatment with ethanol. Electronic Publication     

Dorsal hippocampus, CA3, and CA1 lesions disrupt temporal sequence completion

An experiment was designed to evaluate effects of dorsal hippocampus, dorsal CA3a,b, dorsal CA1, and control lesions on performance of a temporal sequence task. Rats were trained on a sequential learning task involving six spatial locations on a radial 8-arm maze. After initial training followed by surgery, it was found that all lesioned animals were able to remember the sequence. To test temporal sequence completion, rats were started at different positions in the sequence and expected to complete the remainder of the sequence. The results indicate that control rats had no difficulty completing the sequence, regardless of starting point. In contrast, rats with dorsal hippocampus and dorsal CA3a,b lesions made errors by always returning to the first position in the sequence, regardless of which start position was used, whereas rats with dorsal CA1 lesions made random errors in the process of completing the sequence and did not appear to remember the serial order of the spatial sequence. This suggests that the dorsal hippocampus, and specifically the dorsal CA3 in conjunction with CA1, may be involved in temporal pattern completion processes.     

Role of the medial and lateral septum in a variable goal spatial problem solving task.

Rats with lesions to the medial (MS) or lateral septal (LS) nuclei were compared to normal controls (CNT) in the acquisition of a spatial working memory task. In this task, animals were first allowed to explore the unbaited three-table apparatus before being fed on one of the two possible goal tables. Animals were then tested on their ability to return to the table where they just had been fed. Only rats with medial septal damage were clearly impaired on this problem, an impairment that dissipated over days. In contrast, the performance of LS rats was not significantly different from controls. During the second phase of the experiment, the same animals received either atropine sulphate (50 mg/kg, IP), atropine methylnitrate (50 mg/kg, IP), or an equivalent volume of saline. Atropine sulphate produced a sharp decrease in performance by all subjects. Meanwhile, atropine methylnitrate produced a mild temporary deficit only in LS rats. Overall, these results confirm that the medial septum plays a crucial role in the acquisition of problem solving. In addition, these results also suggest that the lateral septum may play a possible role in some form of spatial behavior easily disrupted by atropine methylnitrate.     

Working memory and the aged rat: deficient two-choice win-stay water-escape acquisition and retention

Twelve aged (22 months), 12 adult (12 months) and 12 young (3 months) male Fisher 344 rats were compared on acquisition of a two-choice win-stay (spatial delayed matching-to-sample) water-escape working memory task. The 7 aged, 10 adult and 10 young animals that attained criterion were then compared on performance of the task when the retention interval (RI) was 5, 15, 30, 60 and 120 min. They were also compared on retention after being forced to perform an unsolvable water-escape task in a second maze during the first 5 minutes of a 30-minute RI. The aged animals had longer escape latencies, and required more trials and made more errors in attaining a criterion of 9 correct choices over 10 test trials than did the adult and young animals, who did not differ from one another. Also, comparisons of the animals that attained the acquisition criterion revealed that the aged animals had longer escape latencies and made more errors across all RIs and during trials including an interpolated distraction task than did the adult and young animals. The aged animals failed to perform above chance level on tests at any of the RIs or following the distraction task, while the young animals performed above chance level on tests at all intervals and following distraction. The adult animals failed to perform above chance on tests with the 120-minute RI or following distraction. The aged rats were clearly deficient on acquisition and retention of a two-choice water-escape working memory task.     

Analysis of aversively conditioned learning and memory in rats recovered from pyrithiamine-induced thiamine deficiency

Rats that had recovered from pyrithiamine-induced thiamine deficiency (PTD) were trained on tasks motivated by escape from mild footshock. On postmortem examination, the PTD model showed two consistent lesions: a bilaterally symmetrical lesion of the medial thalamus, which was centered on the internal medullary lamina (IML), and a lesion centered on the medial mammillary nuclei. PTD rats with IML lesions were impaired in learning a spatial nonmatching-to-sample (NMTS) task that was mastered without error by controls and PTD animals without IML lesions. These same animals were able to perform as well as controls on discrimination tasks based on either place or visual (light-dark) cues, although they made more errors than controls in reaching criterion in the initial place discrimination problem. These findings are consistent with findings from appetitively motivated tasks that PTD rats with IML lesions have an impaired capacity for working memory but not for reference memory.     

The effect of early experience on water maze spatial learning and memory in rats

In the first of two experiments on spatial competence, groups of light-reared (LR) and dark-reared (DR) rats were compared using a "latent learning" variation of the Morris Water Maze task. On their initial test, the LR rats benefited more than DR rats did from viewing the room/pool from a platform in the correct location. Further, visually experienced rats remember the location of the platform more than DR rats when retested one month later. In a second experiment, in which a proximal cue as well as location was varied from trial to trial. LR rats again proved to be more competent than their DR counterparts. This second task also revealed significant benefits related to stimulation history in the case of a third group of animals raised in enriched or complex environment (CR) conditions. The results are discussed in terms of the nature of the impact of early experience on the ability to acquire and remember spatial concepts.     

Profound effects of combining choline and piracetam on memory enhancement and cholinergic function in aged rats

In an attempt to gain some insight into possible approaches to reducing age-related memory disturbances, aged Fischer 344 rats were administered either vehicle, choline, piracetam or a combination of choline or piracetam. Animals in each group were tested behaviorally for retention of a one trial passive avoidance task, and biochemically to determine changes in choline and acetylcholine levels in hippocampus, cortex and striatum. Previous research has shown that rats of this strain suffer severe age-related deficits on this passive avoidance task and that memory disturbances are at least partially responsible. Those subjects given only choline (100 mg/kg) did not differ on the behavioral task from control animals administered vehicle. Rats given piracetam (100 mg/kg) performed slightly better than control rats (p<0.05), but rats given the piracetam/choline combination (100 mg/kg of each) exhibited retention scores several times better than those given piracetam alone. In a second study, it was shown that twice the dose of piracetam (200 mg/kg) or choline (200 mg/kg) alone, still did not enhance retention nearly as well as when piracetam and choline (100 mg/kg of each) were administered together. Further, repeated administration (1 week) of the piracetam/choline combination was superior to acute injections. Regional determinations of choline and acetylcholine revealed interesting differences between treatments and brain area. Although choline administration raised choline content about 50% in striatum and cortex, changes in acetylcholine levels were much more subtle (only 6–10%). No significant changes following choline administration were observed in the hippocampus. However, piracetam alone markedly increased choline content in hippocampus (88%) and tended to decrease acetylcholine levels (19%). No measurable changes in striatum or cortex were observed following piracetam administration. The combination of choline and piracetam did not potentiate the effects seen with either drug alone, and in certain cases the effects were much less pronounced under the drug combination. These data were discussed as they relate to possible effects of choline and piracetam on cholinergic transmission and other neuronal function, and how these effects may reduce specific memory disturbances in aged subjects. The results of these studies demonstrate that the effects of combining choline and piracetam are quite different than those obtained with either drug alone and support the notion that in order to achieve substantial efficacy in aged subjects it may be necessary to reduce multiple, interactive neurochemical dysfunctions in the brain, or affect activity in more than one parameter of a deficient metabolic pathway.