Non-spatial water radial-arm maze learning in mice

Recently, we published a method for examining working and reference memory in mice using a spatial version of the water radial-arm maze. Here we describe a non-spatial version of the same maze. BXSB mice were able to learn the maze as shown by the decrease in the number of working and reference memory errors over sessions. This maze was used to examine learning differences between males and females and between mice with misplaced clusters of neurons in layer I of cortex (ectopias) and those without. In a prior study using the spatial version of the water radial-arm maze, male BXSB mice had poorer working memory than females during the acquisition phase. Similarly, in this study male BXSB mice demonstrated impaired working memory during the asymptotic phase of non-spatial radial-arm maze learning. Two prior studies showed that mice with neocortical ectopias demonstrated working memory impairments compared to non-ectopic littermates in the spatial version of the water radial-arm maze. Contrary to this, in the non-spatial radial-arm maze used here, ectopic mice were not impaired in working memory and showed better memory when the working memory 'load' was the highest. Overall, both versions of the maze can be useful tools to assess spatial and non-spatial working and reference memory in mice.     

Impaired water maze learning performance in mu-opioid receptor knockout mice

Previous study has demonstrated that the lack of mu-opioid receptor decreased LTP in the dentate gyrus of the hippocampus, suggesting the possibility that the lack of mu-opioid receptor may accompany a change in learning and memory. However, no behavioral study has been undertaken to correlate LTP deficits with spatial memory impairment in mu-opioid receptor knockout mice. Therefore, the present study investigated the hypothesis that mu-opioid receptors contribute to learning and memory by using the Morris water maze, and comparing responses in wild type and mu-opioid receptor gene knockout mice. Our results indicated that mu-opioid receptor knockout mice showed a significant spatial memory impairment compared to wild type in the Morris water maze. This result suggests that the expression of mu-opioid receptor plays an important role in spatial learning and memory examined by Morris water maze.     

The olfactory tubing maze: a new apparatus for studying learning and memory processes in mice

In order to have an ethologically relevant behavioral task, we developed the olfactory tubing maze to study learning and memory processes in mice. Mice have to make two olfactory-reward associations across three training sessions. The maze is made up of four identical testing chambers connected to each other by semicircular cylinders. After having chosen one of two odors presented on each side of a testing chamber, the mice have to run to the next testing chamber. From one testing chamber to the next, the side for presentating each odor is randomly assigned. The mouse must run through the entire circular maze to make a response at the four testing chambers. A complete session consists of 20 trials made by running five times clockwise through the maze with 4 trials per run. The training and data recording are fully automated by a custom-made software program. Three different experiments were performed. The results indicated that mice can easily make the olfactory discriminative associations in this new apparatus. Analysis of the data suggests that it would be possible using this olfactory tubing maze to study sub-categories of memory similar in some respects to those observed in humans. Consequently, possible effects on learning and memory of classical treatments (i.e. pharmacological or lesions) or genetic modifications in transgenic or gene-targeting mice could be tested.     

SR 141716A prevents delta 9-tetrahydrocannabinol-induced spatial learning deficit in a Morris-type water maze in mice

This study reports a series of spatial discrimination procedures in a Morris-type maze to investigate the effects of delta9-tetrahydrocannabinol (delta9-THC) on different phases of learning and memory in mice. Adult male mice were given training trails to find the submerged platform at a fixed location in the water maze adapted for mice. In additional experiments, mice were trained with the repeated acquisition procedure to test the working memory. Results indicate that delta9-THC (8 mg/kg i.p.) 30 min pretest impaired specifically the acquisition of spatial learning and the performance of mice in the working memory task, while consolidation and retrieval of a previously learned task were not affected. There was no evidence of motoric difficulty, as the number of quadrant line crossings was not decreased and no visible sign of sensorimotor disturbance was observed during swimming. Pretreatment with SR 141716A (1 mg/kg i.p.), a CB1 cannabinoid receptor antagonist, significantly prevented the learning deficits in the water maze. These findings show that delta9-THC impairs spatial discrimination learning in a selective way in the water maze in mice and that these deficits may be mediated by cannabinoid receptors.     

Parkinson's disease patient's behaviour in a covered maze learning task

A computerized maze task was constructed that allowed only partial vision of the maze structure and produced measurements for separate analysis of cognitive processes described as impaired in Parkinson's disease. Eighteen patients suffering from mild Parkinson's disease and 18 individually matched normal controls were investigated. Baseline task response times were found to be identical for both groups. Differences between patients' and controls' performance could be related to (a) a response bias in Parkinson patients that favoured repetition of the previous action and slowed down shifting and (b) an impairment of multistep plan generation. It is speculated that the response bias reflects the disinhibition of cortico-thalamo-cortical reverberation loops which results from striatal dopamine depletion.     

Hippocampal synaptic depression following spatial learning in a complex maze

Activity-dependent alteration in synaptic efficacy is referred to as synaptic plasticity and is the believed hallmark of any learning process. Here we employed a recently validated complex maze for spatial training and investigated the impact of repeated and extensive training on basal synaptic transmission of the hippocampal Schaffer collateral-CA1 synaptic connection in vitro. In the present experiments, male CD-1 mice were trained in a complex maze for eight consecutive days five times a day. Subsequently, input-output functions of field excitatory postsynaptic potentials (fEPSPs) recorded in the CA1 area following stimulation of the Schaffer collateral-commissural fiber pathway were analyzed in acute hippocampal slices. We found a marked right shift of the fEPSP response in trained compared to untrained animals while fiber volley size remained unchanged. The findings provide evidence for a direct implication of homosynaptic hippocampal long-term depression in a spatial learning paradigm.     

Validation of a 2-day water maze protocol in mice

We present a 2-day water maze protocol that addresses some of potential confounds present in the water maze when using the aged subjects typical of studies of neurodegenerative disorders, such as Alzheimer's disease. This protocol is based on an initial series of training trials with a visible platform, followed by a memory test with a hidden platform 24h later. We validated this procedure using aged (15-18m) mice expressing three Alzheimer's disease-related transgenes, PS1(M146 V), APP(Swe), and tau(P301L). We also tested these triple transgenic mice (3xTG) and age and sex-matched wild-type (WT) in a behavioral battery consisting of tests of motor coordination (balance beam), spatial memory (object displacement task) visual acuity (novel object recognition task) and locomotor activity (open field). 3xTG mice had significantly longer escape latencies in the memory trial of the 2-day water maze test than WT and than their own baseline performance in the last visible platform trial. In addition, this protocol had improved sensitivity compared to a typical probe trial, since no significant differences between genotypes were evident in a probe trial conducted 24h after the final training trial. The 2-day procedure also resulted in good reliability between cohorts, and controlled for non-cognitive factors that can confound water maze assessments of memory, such as the significantly lower locomotor activity evident in the 3xTG mice. A further benefit of this method is that large numbers of animals can be tested in a short time.     

Prefrontal cortical mediation of rats' place learning in a modified water maze

The acquisition of a place learning task in a water maze modified from the “standard” set-up by restriction of distal cues and addition of “proximal” cues (ping-pong balls in fixed positions on the surface of the water) was tested in three groups of rats: (I) animals subjected to bilateral ablation of the anteromedial prefrontal cortex, (II) rats in which the parietal “association” cortex had been removed bilaterally, and (III) a sham operated control group. The task acquisition of the prefrontaly ablated group was significantly impaired, whereas the animals in which the parietal cortex had been removed acquired the task as quickly as the control group. Upon reaching criterion level performance all animals were tested on “challenge” sessions on which the cues were manipulated. Such “challenges” demonstrated that the animals of all three groups discriminated between the distal cues and utilized such a discrimination for navigational purposes.     

Estradiol to aged female or male mice improves learning in inhibitory avoidance and water maze tasks

Although 17beta-Estradiol (E2) improves cognitive performance of aged female mice, its mnemonic effects when administered post-training to aged male mice have not been examined. E2 (10 microg, SC) or oil vehicle was administered to intact, 24-month-old female or male congenic (primarily C57BL/6 background) mice immediately after training in the inhibitory avoidance or water maze tasks. Following behavioral testing, effects of 1 or 24 h of E2 exposure on hippocampal levels of E2 and brain-derived neurotrophic factor (BDNF) were examined. Female and male mice administered E2 showed significantly better performance in the inhibitory avoidance task than did vehicle-administered mice. When tested 24 h after training, mice that received E2 had significantly longer latencies to cross-over to the shock-associated side of the chamber than did vehicle-administered mice. Female or male mice administered E2 showed significantly better performance in the reference memory aspect of the spatial water maze task. When tested 30 min after training, mice administered E2 had shorter latencies to, and spent longer swimming in, the quadrant that the hidden platform had previously been located in. E2 administration produced physiological levels of E2 in the hippocampus 1 and 24 h after E2. BDNF levels in the hippocampus were decreased following 1 h of E2 exposure compared to vehicle. These findings suggest that E2 to female and male mice may overcome age-related deficits in reference memory in an emotional or spatial learning task.     

Impaired water maze learning performance without altered dopaminergic function in mice heterozygous for the GDNF mutation

Exogenous glial cell line-derived neurotrophic factor (GDNF) exhibits potent survival-promoting effects on dopaminergic neurons of the nigrostriatal pathway that is implicated in Parkinson's disease and also protects neurons in forebrain ischemia of animal models. However, a role for endogenous GDNF in brain function has not been established. Although mice homozygous for a targeted deletion of the GDNF gene have been generated, these mice die within hours of birth because of deficits in kidney morphogenesis, and, thus, the effect of the absence of GDNF on brain function could not be studied. Herein, we sought to determine whether adult mice, heterozygous for a GDNF mutation on two different genetic backgrounds, demonstrate alterations in the nigrostriatal dopaminergic system or in cognitive function. While both neurochemical and behavioural measures suggested that reduction of GDNF gene expression in the mutant mice does not alter the nigrostriatal dopaminergic system, it led to a significant and selective impairment of performance in the spatial version of the Morris water maze. A standard panel of blood chemistry tests and basic pathological analyses did not reveal alterations in the mutants that could account for the observed performance deficit. These results suggest that endogenous GDNF may not be critical for the development and functioning of the nigrostriatal dopaminergic system but it plays an important role in cognitive abilities.     

Spatial learning and memory, maze running strategies and cholinergic mechanisms in two inbred strains of mice

The acquisition process of the radial maze task was studied in two inbred strains of mice, C57BL/6 and DBA/2. A quantitative and qualitative evaluation of performance was performed and the pretest level of activity was measured. The results showed a significant correlation between activity and performance since the highly active C57BL/6 mice exhibited better performance of the radial maze task than the less active DBA/2 mice. Moreover, for correct trials, strain-dependent maze-running strategies were observed: while both strains displayed about the same percentage of clockwise and spatial strategies, it was observed that among the spatial strategies C57BL/6 used a larger number of different correct solutions. Subsequently, the effect of scopolamine administration on working memory processes was assessed in sequential and discrete trials. A different reactivity of each strain to anti-cholinergic treatment was found in discrete trials since only DBA/2 mice were impaired. The effect of scopolamine is discussed in relation to the different models of information processing involved in learning and memorizing the experimental rule.     

Allopregnanolone inhibits learning in the Morris water maze

The progesterone metabolite allopregnanolone (3alpha-OH-5alpha-pregnane-20-one) inhibits neural functions, enhancing the GABA induced GABA(A) receptor activation. This effect is benzodiazepine like and benzodiazepines are known to impair memory. Acute effects of allopregnanolone on the hippocampus dependent spatial learning in the Morris water maze have not been studied. Adult male Wistar rats where injected (i.v.) with allopregnanolone (2 mg/kg), or vehicle, daily for 11 days. At 8 or 20 min after each injection, studies of place navigation were performed in the Morris water maze. Allopregnanolone concentrations in plasma and in nine different brain areas where analyzed by radioimmunoassay. The latency to find the platform was increased 8 min after the allopregnanolone injection, while normal learning was seen after 20 min. Swim speed did not differ between groups. A higher number of rats were swimming close to the pool wall (thigmotaxis) in the 8 min allopregnanolone group compared to the other groups. Allopregnanolone concentrations in the brain tissue at 8 min were 1.5 to 2.5 times higher then at 20 min after the allopregnanolone injections. After vehicle injections the brain concentrations of allopregnanolone were at control levels. Plasma concentrations of allopregnanolone followed the same pattern as in the brain, with the exception of an increase 8 min after vehicle injections. The natural progesterone metabolite allopregnanolone can inhibit learning in the Morris water maze, an effect not caused by motor impairment. The learning impairment might be due to a combination of changed swimming behavior and difficulties in navigation.     

Effects of spaced learning in the water maze on development of dentate granule cells generated in adult mice

New dentate granule cells (GCs) are generated in the hippocampus throughout life. These adult‐born neurons are required for spatial learning in the Morris water maze (MWM). In rats, spatial learning shapes the network by regulating their number and dendritic development. Here, we explored whether such modulatory effects exist in mice. New GCs were tagged using thymidine analogs or a GFP‐expressing retrovirus. Animals were exposed to a reference memory protocol for 10–14 days (spaced training) at different times after newborn cells labeling. Cell proliferation, cell survival, cell death, neuronal phenotype, and dendritic and spine development were examined using immunohistochemistry. Surprisingly, spatial learning did not modify any of the parameters under scrutiny including cell number and dendritic morphology. These results suggest that although new GCs are required in mice for spatial learning in the MWM, they are, at least for the developmental intervals analyzed here, refractory to behavioral stimuli generated in the course of learning in the MWM. © 2015 Wiley Periodicals, Inc.     

Long-term effect of glucocorticosteroids on neuromuscular blocking in mice.

The effect of dexamethasone, given thrice weekly, on neuromuscular blocking by D-tubocurarine was studied in mice. Dexamethasone antagonised the effect of D-tubocurarine. This antagonism developed gradually during the first two weeks of treatment. The possible relationship between this finding and the beneficial effect of glucocorticosteroids in patients with myasthenia gravis is discussed.     

Effects of neostriatal and mesocorticolimbic dopamine depletions on learning in a water maze

Females born in all-female litters and females stemming from predominantly male litters were compared for masculine (mounting) and feminine sexual behavior (lordosis) under various dosages of estradiol followed by progresterone and under testosterone stimulation. Estradiol did not affect mounting levels in these groups. A critical dose level was established at 0.25 mg/animal of estradiol for both groups with respect to receptive behavior. Testosterone treatment reliably stimulated mounting in these females, but no differences were determined with respect to the litter composition in utero. The results are considered as a critical and negative finding with regard to the possibility that testosterone stemming from male littermates in utero affects the female's sexual development.     

Sex differences in visual-spatial learning using a virtual water maze in pre-pubertal children

Gonadal steroid effects during puberty are often hypothesized to account for the male advantage seen in certain spatial tasks. One spatial task where males consistently show better performance than females is the Morris Water Task in which subjects must navigate to a goal location in a pool. We examined whether sex differences exist in pre-pubertal children completing a Virtual Morris Water Task, which has previously shown strong sex differences in adults. Pre-pubertal boys show superior performance to similar-aged girls, as evidenced by shorter latencies to find the platform and stronger preferences for the platform location during a probe trial. These results suggest that sex differences in spatial learning and memory exist prior to puberty and do not appear to require the effects of sex hormones at puberty. Rather, these differences may reflect early-life hormonal effects on hippocampal-dependent processes and may suggest different preferential learning strategies by boys and girls.     

Age-related changes in radial-arm maze learning and basal forebrain cholinergic systems in senescence accelerated mice (SAM).

Age-related changes in learning performance and the brain cholinergic system were studied in a senescence accelerated mice-prone series (SAM-P/8) and a senescence accelerated mice-resistant series (SAM-R/1, control) bred under specific pathogen-free conditions. In a radial-arm maze task, SAM-P/8 mice at 4 and 12 months of age showed virtually no significant impairment in working memory or reference memory compared with SAM-R/1 mice at the same age, although they needed more time to complete a trial than SAM-R/1. In contrast, in a passive avoidance task, SAM-P/8 showed a marked age-accelerated deficit in acquisition performance relative to SAM-R/1. Also, SAM-P/8 showed an age-accelerated decrease in locomotion and rearing in an open-field box. At the end of these behavioral tasks, neurochemical analyses showed that there were no differences in the concentrations of acetylcholine (ACh) in the cortex, hippocampus, striatum, midbrain, or cerebellum between SAM-P/8 and SAM-R/1. Although SAM-P/8 mice did not demonstrate any age-accelerated decline in radial-arm maze performance, they showed a normal age-related decline particularly in working memory, equal to that observed in SAM-R/1. Also, ACh levels in the aged groups of SAM-P/8 showed a significant decrease related to normal aging in the hippocampus and striatum, and a slight decrease in the cortex compared to the young group of the same strain. Thus, we found that SAM-P/8 show dissociative effects of aging in spatial learning and passive avoidance performance.     

Stress impairs performance in spatial water maze learning tasks

The water maze task has been developed to test spatial learning abilities in rats or mice, and is widely used. Though it has been reported before that numerous cognitive abilities are of importance for learning this task, poor performance is usually interpreted as an impairment of spatial memory formation. Previous investigations that tried to correlate long-term potentiation (LTP) of synaptic transmission with spatial learning abilities in rats reported that injection of drugs or specific gene deletions which blocked the expression of LTP correlated with learning impairments of spatial tasks in a water maze. Recent studies, however, have shown that pretraining enables these animals to learn such spatial tasks even though LTP was still found to be blocked. I investigated to what degree altered fear condition and stress perception could account for the impaired spatial learning when no pretraining is given. In a fear habituation task, unhandled rats preferred a dark over a well lit chamber more than handled animals did, but unhandled rats favoured the lit chamber more in an active avoidance task. They also performed poorly in a spatial water maze task compared with handled rats. Rats pretrained in a radial arm maze performed better in a water maze than non-pretrained rats. No difference between groups was found in a non-spatial water maze task. On the other hand, when pretrained in a water maze, rats performed only marginally better in a radial arm maze compared to non-pretrained animals. Since animals have to be handled to learn a radial arm maze, the difference in this task was not due to stress but most probably due to getting accustomed to the room dimensions prior to learning the spatial task. The results suggest that impaired learning of spatial tasks in the water maze can be due to increased stress and decreased fear conditioning without actually affecting spatial learning abilities. These results question the interpretations of the results of some previously published results of spatial water maze tasks.