Sexually dimorphic responses to neonatal basal forebrain lesions in mice: I. Behavior and neurochemistry

The nucleus basalis magnocellularis (nBM) provides the primary source of cholinergic input to the cortex. Neonatal lesions of the nBM produce transient reductions in cholinergic markers, persistent abnormalities in cortical morphology, and spatial navigation impairments in adult mice. The present study examined sex differences in the effects of an electrolytic nBM lesion on postnatal day 1 (PND 1) in mice on behavior and neurochemistry in adulthood. Mice were lesioned on PND 1 and tested at 8 weeks of age on a battery of behavioral tests including passive avoidance, cued and spatial tasks in the Morris water maze, simple and delayed nonmatch to sample versions of an odor discrimination task, and locomotor activity measurements. Following behavioral testing, mice were sacrificed for either morphological assessment or neurochemical analysis of a cholinergic marker or catecholamines. There were no lesion or sex differences in acquisition or retention of passive avoidance, performance of the odor discrimination tasks, or activity levels. Control mice showed a robust sex difference in performance of the spatial water maze task. The lesion produced a slight cued but more dramatic spatial navigation deficit in the water maze which affected only the male mice. Neurochemical analyses revealed no lesion-induced changes in either choline acetyltransferase activity or levels of norepinephrine or serotonin at the time of testing. The subsequent report shows a sex difference in lesion-induced changes in cortical morphology which suggests that sexually dimorphic cholinergic influences on cortical development are responsible for the behavioral deficits seen in this study. © 1998 John Wiley & Sons, Inc. J Neurobiol 37: 582–594, 1998     

The ACTH/MSH (4-9) analog Org2766 improves retrieval of information after a fimbria fornix transection

The fimbria fornix of male Wistar rats was transected unilaterally after they had been successfully trained in the Morris maze and the passive avoidance task. Sham-operated and lesioned animals were treated either with Org2766 or saline for two weeks. Subsequently, the performance of all groups was tested again starting two days after the last treatment. The lesioned animals showed a deficit in performance in both tasks, indicating interference of the lesion with retrieval of information. Org2766 improved the poor performance of the lesioned animals in the Morris maze, but not in the passive avoidance task.     

The hippocampus and flexible spatial knowledge in rats

Lesions to the hippocampal system in rats result in a profound impairment of place or locale spatial learning although other learning strategies remain unaltered. The main objective of the present study was to investigate whether the spatial knowledge preserved in the hippocampal animals can be expressed flexibly under conditions different from those of the acquisition period. Rats with neurotoxic lesions to the dorsal hippocampus and sham-operated subjects were trained to reach the goal arm in a four-arm plus-shaped maze using a constant starting arm. During the training a transparent plexiglas barrier divided the maze in two equal halves in such a way that the animals could only travel from the starting arm to the goal arm, not having access to the remaining 50% of the maze. After seven days of training, a transfer test was used in which the starting arms were the two arms from which the animals had not started during the training phase. Results indicated that the lesioned rats made significantly more errors than the control subjects. But the most interesting results revealed that the kind of error made by the lesioned animals was congruent with the turn that they had to make during the acquisition phase in order to access the goal arm (reinforced). These results suggest that when the hippocampus is damaged a preserved highly inflexible egocentric strategy is employed to solve the spatial problem.     

Dihydrotestosterone modulates spatial working-memory performance in male mice

Androgens affect cognitive processes in both humans and animals. The effects of androgens may be limited to certain cognitive domains, specifically spatial memory, but this hypothesis remains elusive. Here, we tested castrated and sham-operated mice in various behavioral tasks to ask whether androgens affect multiple or specific cognitive domains in male mice. Castration impaired spatial working memory performance in the delayed matching to place water maze task following a 1-h, but not a 1-min, retention interval, as has been reported for rats. In contrast, castration had no effect on novel object recognition memory, spatial reference memory in the water maze, motor coordination, or passive avoidance memory. Castration increased anxiety-like behavior in the open field test, but not the elevated zero maze. Finally, we assessed the effects of androgen replacement with non-aromatizable dihydrotestosterone on spatial working memory following various retention intervals. Dihydrotestosterone recovered spatial memory performance following a 24-h, but not a 1-h retention interval, and had no effect at other retention intervals. These data support that in male mice androgens specifically affect spatial working memory performance, and that the neurobiological processes underlying spatial memory formation may be differentially affected by androgens.     

Lesions of the Basal Forebrain Cholinergic System in Mice Disrupt Idiothetic Navigation

Loss of integrity of the basal forebrain cholinergic neurons is a consistent feature of Alzheimer’s disease, and measurement of basal forebrain degeneration by magnetic resonance imaging is emerging as a sensitive diagnostic marker for prodromal disease. It is also known that Alzheimer’s disease patients perform poorly on both real space and computerized cued (allothetic) or uncued (idiothetic) recall navigation tasks. Although the hippocampus is required for allothetic navigation, lesions of this region only mildly affect idiothetic navigation. Here we tested the hypothesis that the cholinergic medial septo-hippocampal circuit is important for idiothetic navigation. Basal forebrain cholinergic neurons were selectively lesioned in mice using the toxin saporin conjugated to a basal forebrain cholinergic neuronal marker, the p75 neurotrophin receptor. Control animals were able to learn and remember spatial information when tested on a modified version of the passive place avoidance test where all extramaze cues were removed, and animals had to rely on idiothetic signals. However, the exploratory behaviour of mice with cholinergic basal forebrain lesions was highly disorganized during this test. By contrast, the lesioned animals performed no differently from controls in tasks involving contextual fear conditioning and spatial working memory (Y maze), and displayed no deficits in potentially confounding behaviours such as motor performance, anxiety, or disturbed sleep/wake cycles. These data suggest that the basal forebrain cholinergic system plays a specific role in idiothetic navigation, a modality that is impaired early in Alzheimer’s disease.     

Foetal amygdalar transplantation facilitates recovery of retention deficit in CeA lesioned rats

Neural tissue transplant has come off age as a valuable technique for studying normal development and regeneration. Bilateral lesions of the central nucleus of amygdala (CeA) produce complete retention and acquisition deficit in inhibitory avoidance paradigms. The present study reports recovery of retention deficit in active avoidance task (AA) after amygdalar tissue transplantation in CeA lesioned rats. In a group of adult wistar rats, bilateral lesions of the CeA were produced electrolytically. In a separate group of rats foetal amygdalar tissue was transplanted at the CeA lesioned site 2 days after producing lesion. All the rats were trained on AA task before and after 5 days of lesion. In bilaterally CeA lesioned rats, the percentage of avoidance (% avoidance) decreased significantly (P < 0.05) from 85 +/- 18% prelesion to 15.5 +/- 35% postlesion. However, no change in the % avoidance was observed after amygdalar tissue transplantation. The results indicate that the transplanted rats are capable of retaining the learnt information in contrast to the lesion alone group of rats.     

Comparison of neurokinin SP with diazepam in effects on memory and fear parameters in the elevated T-maze free exploration paradigm.

The elevated T-maze was combined with a free exploration protocol, which, in contrast to the conventional procedure, dispenses with handling of the animals during the experimental sessions. This allows measurement of fear indexes derived from the elevated plus-maze as well as assessment of acquisition of open arm avoidance and open arm escape in one continuous session. Retention of the different fear-responses is measured 72 h later without drug treatment. In order to assess the effects of two known anxiolytics in this paradigm, rats received an IP injection of diazepam (1 to 4 mg/kg), substance P (5 to 500 microg/kg) or vehicle (1 ml/kg) and were tested on the T-maze for 5 min. Diazepam elevated open arm activity, indicative of an anxiolytic effect. The drug also increased the latency to escape from the open arms, but did not significantly affect acquisition of open arm avoidance. During the retention trial, diazepam in higher doses impaired the performance of both fear-responses, suggestive of an anterograde amnesic effect. Substance P did not influence acquisition and retention of open arm avoidance and escape. However, in high doses, the peptide increased the sojourn time in the central arena of the maze, indicating reduced fear and, hence, a dissociation between anxiolytic and amnesic effects. The present findings demonstrate that the elevated T-maze free exploration paradigm is sensitive to anxiolytic and memory-modulating effects of drugs.     

Low-level exposure to pulsed 900 MHz microwave radiation does not cause deficits in the performance of a spatial learning task in mice

There is some concern that short-term memory loss or other cognitive effects may be associated with the use of mobile cellular telephones. In this experiment, the effect of repeated, acute exposure to a low intensity 900 MHz radiofrequency (RF) field pulsed at 217 Hz was explored using an appetitively-motivated spatial learning and working memory task. Adult male C57BL/6J mice were exposed under far field conditions in a GTEM cell for 45 min each day for 10 days at an average whole-body specific energy absorption rate (SAR) of 0.05 W/kg. Their performance in an 8-arm radial maze was compared to that of sham-exposed control animals. All behavioral assessments were performed without handlers having knowledge of the exposure status of the animals. Animals were tested in the maze immediately following exposure or after a delay of 15 or 30 min. No significant field-dependent effects on performance were observed in choice accuracy or in total times to complete the task across the experiment. These results suggest that exposure to RF radiation simulating a digital wireless telephone (GSM) signal under the conditions of this experiment does not affect the acquisition of the learned response. Further studies are planned to explore the effects of other SARs on learned behavior. Bioelectromagnetics 21:151-158, 2000. Published 2000 Wiley-Liss, Inc.     

Effect of disruption of the entopeduncular nucleus on rat behavior in simultaneous discrimination and open field tests]

The influence of unilateral destruction of n. entopeduncularis on learning and retention of simultaneous discrimination in a T-maze and adaptation in the open filed was studied in 29 adult rats. Destruction of n. entopeduncularis in rats resulted in appreciable deviations from the normal behavior during realization of earlier acquired simultaneous discrimination and slowing down of the acquisition of a new habit. Differences in behavioral activity of the lesioned animals in the maze and open field were revealed.     

6-Hydroxydopamine infusions into the structures of mesolimbic dopaminergic system alter the memory enhancing effect of CK-8US and caerulein in rats

The influence of bilateral destruction of dopaminergic endings in the anterior and in the posterior part of nucleus accumbens (NAS) and in the nucleus septi lateralis (NSL), by 6-hydroxydopamine (6-OHDA) infusions, on the facilitatory effect of cholecystokinin-unsulfated octapeptide (CCK-8US) and caerulein (CER) on memory motivated affectively was investigated in male Wistar rats. CCK-8US and CER were given s.c. at the doses of 10 microg/kg and 0.5 microg/kg respectively, immediately after a single learning trial in a passive avoidance situation, ten days after bilateral 6-OHDA lesions (desipramine pre-treatment; 25 mg/kg, i.p.) of these structures. Bilateral 6-OHDA lesions to the anterior and to the posterior part of NAS totally abolished and significantly attenuated, respective, the facilitatory effect of CCK-8US and CER on retention of a passive avoidance behaviour evaluated 24 h later, while bilateral lesions to NSL did not have any influence on it. Moreover, neither, destruction of dopaminergic endings in lesioned structures, nor application of CCK-8US and CER changed the spontaneous psychomotor activity of rats estimated in an "open field" test. These results may indicate that dopaminergic projection to the anterior part of NAS is mainly responsible for the facilitatory effect of CCK-8US and CER on memory motivated affectively.     

Exposure to stable flies reduces spatial learning in mice: involvement of endogenous opioid systems

Abstract. Biting flies influence both the physiology and behaviour of domestic and wild animals. This study demonstrates that relatively brief (60min) exposure to stable flies, Stomoxys calcitrans (L.), affects the spatial abilities of male mice. Stable fly exposure resulted in poorer subsequent performance in a water maze task in which individual mice had to learn the spatial location of a submerged hidden platform using extramaze visual cues. Determinations of spatial acquisition and retention were made with mice that had been previously exposed for 60min to either stable flies or house flies, Musca domestica (L.). Mice exposed to stable flies displayed over one day of testing (six blocks or sets of four trials) significantly poorer acquisition and retention of the water maze task than either mice that had been exposed to house flies or fly-naive mice. This attenuation of spatial learning occurred in the absence of any evident sensorimotor or motivational impairments. The reduction in spatial abilites involved endogenous opioid systems, as the decreased performance resulting from stable fly exposure was blocked by pre-treatment with the prototypic opiate antagonist, naltrexone. These results indicate that relatively brief exposure to biting flies can lead to a decrease in spatial abilities which is associated with enhanced endogenous opioid activity. These results support the involvement of endogenous opioid systems in the mediation of the behavioural and physiological effects of biting fly exposure. They further suggest that decreases in spatial abilities and performance may be part of the behavioural consequences of biting fly exposure in domestic and wild animals.     

Effect of circadian phase on performance of rats in the Morris water maze task

The authors examined spatial working memory in the Morris water maze during the activity and rest periods of Wistar rats. Wheel-running activity was measured continuously as a marker of circadian phase. To minimize possible masking effects on performance, animals were placed in constant dim light the day before testing and tested in similar light conditions. Three experiments were run, each of them using animals varying in their previous experience in the water maze. Half of the animals of each experiment were tested 2 to 3 h after activity onset (active group), and the other half were tested 14 to 15 h after activity onset (inactive group). In the three experiments, a significant phase effect was observed in the animals' performance in the water maze; animals tested in the active phase showed steeper acquisition curves. These phase effects on performance are due to the animals' search pattern and not to a better acquisition and maintenance of spatial information; rats tested in the inactive phase found the platform faster on the first trial of the test, when the information on the location of the platform had not been presented to the animals. This effect vanished as the amount of training in the pool increased. Finally, swimming speed also showed a temporal effect, suggesting the existence of a phase effect for motivation to escape from the water; rats tested during their inactive phase tended to swim faster. All together, the data suggest a modulating effect of the biological clock on performance in the water maze, particularly when the animals are less experienced.     

Effects of a memory enhancing peptide on cognitive abilities of brain-lesioned mice: additivity with huperzine A and relative potency to tacrine.

Alzheimer's disease (AD) and related dementing disorders having cognitive manifestations represent an increasing threat to public health. In the present study, the effects of a memory enhancing NLPR tetra-peptide (MEP), huperzine A (Hup A), or a combination of the two on the cognitive abilities of brain-lesioned mice were evaluated and compared with tacrine in the passive avoidance and Y-water maze tests for the acquisition and retention aspects of cognitive functions. MEP at microg kg(-1) doses, and Hup A or tacrine at mg kg(-1) doses significantly reversed the cognition deficits induced by scopolamine. For acquisition ability, it was observed that mice administered with MEP (4.0 microg kg(-1)) spent less time escaping onto the platform in the water maze than those treated with tacrine (1.5 mg kg(-1)); whereas for memory retention, tacrine-administration resulted in a higher step-through latency in mice at the tested dose regime. In addition, co-administration of MEP (2.0 microg kg(-1)) and Hup A (0.1 mg kg(-1)) exhibited an additive effect resulting in considerable improvements in both acquisition and retention abilities of brain-lesioned mice. The results demonstrated that MEP was highly efficient in the rescue of cognitive abilities of brain-lesioned mice and in particular, the effective doses of MEP were about two orders of magnitude lower than that of tacrine, a therapeutic currently used in the treatment of AD. Moreover, MEP and Hup A were effective at reduced doses when the two were co-administered, providing a rationale for their combined usage in the treatment of cognitive deficits.     

Spatial learning in deer mice: sex differences and the effects of endogenous opioids and 60 Hz magnetic fields

We examined the effects of brief exposure to weak 60 Hz extremely low frequency (ELF) magnetic fields and opioid systems on spatial behavior and learning in reproductive adult male and female deer mice, Peromyscus maniculatus. Sex differences were evident in spatial performance, with male deer mice displaying significantly better performance than female mice in the Morris water maze, whereby animals had to acquire and retain the location of a submerged hidden platform. Brief (maximum 5 min) exposure to weak (100 T) 60 Hz magnetic fields during task acquisition significantly improved female performance, eliminating the sex differences in acquisition. The opiate antagonist, naltrexone, also improved female acquisition, though significantly less than the magnetic fields. These facilitatory effects involved alterations of non-spatial (task familiarization and reduction of related anxiety/aversive related behaviors) and possibly spatial aspects of the task. Enhancement of enkephalin activity with the enkephalinase inhibitor, SCH 34826, significantly reduced task performance by male deer mice. Both naltrexone and the 60 Hz magnetic fields attenuated the enkephalin mediated reductions of spatial performance. These findings indicate that brief exposure to 60 Hz magnetic fields can enhance water maze task acquisition by deer mice and suggest that these facilitatory effects on spatial performance involve alterations in opioid activity.Abbreviations ELF extremely low frequency - Hz hertz     

Effects of bilateral electrolytic lesions of the medial nucleus accumbens on exploration and spatial learning.

Rats with electrolytic lesions of the medial part of the nucleus accumbens, comprising the shell region, were compared to sham-operated rats in tests of exploration in a T-maze, in a hole-board, and in an elevated (+)-maze and in a test of water maze spatial learning. Rats with medial nucleus accumbens lesions had higher choice latencies than sham-operated controls during the beginning of the spontaneous alternation test. A higher number of hole pokes was found in the lesioned group, but only during the beginning of the second day of testing. In the elevated (+)-maze, lesioned rats had a higher number of closed and total arm entries and spent more time in the center region. The lesioned group did not differ from the control group for the number of alternations in the T-maze, for horizontal and vertical motor activity in the hole-board, and for acquisition or reversal of spatial learning in the Morris water maze. These results indicate that lesions of the medial nucleus accumbens slowed down decision time during spontaneous alternation testing and increased exploration in a time and test-specific manner without altering acquisition of a reference memory task.     

Behavior strategy learning in rat: effects of lesions of the dorsal striatum or dorsal hippocampus

Depending on task demands, there is a growing body of evidence suggesting that the dorsal striatum plays a critical role in not only learning new response strategies but also in the inhibition of pre-existing strategies when a shift in strategy is required. The present experiment examined the effects of lesions of the dorsal striatum or dorsal hippocampus on acquisition of a response-learning rule and a place-learning rule in a Greek Cross version of the Morris water maze. Specifically, adult Long-Evans rats were prepared with either sham lesions or lesions to one of two subcortical areas of the brain considered necessary for processing nondeclarative or declarative memories, the dorsal striatum or the hippocampus, respectively. An analysis of the trial 2 performance pooled across reversals revealed hippocampus lesions induced accelerated acquisition when a response-learning rule was required. A much smaller enhancement effect was observed in dorsal striatum-lesioned animals in the place-learning paradigm. Dorsal hippocampus- and dorsal striatum-lesioned animals were highly impaired on place learning and response learning, respectively. The present results are congruent with a growing body of literature suggesting that different anatomical substrates are involved in the acquisition and maintenance of different types of information, that these processes can occur simultaneously and in parallel, and that the dorsal striatum is necessary for the mediation of stimulus-response learning, while the hippocampus is necessary to mediate the expression of place learning.     

Static magnetic field exposure affects behavior and learning in rats

The present work investigated the behavioral effects of a moderate exposure (1 h per day for 5 consecutive days) to a static magnetic field (SMF, 128 mT) in male rats. SMF effects were evaluated in two sets of control and SMF-exposed rats. One set of animals was used for evaluation of SMF potential effects on emotional behaviors in the elevated plus maze and in the open field. The other set of animals was tested for learning and memory abilities in different procedures of the Morris water maze task. We found no significant difference between control and SMF-exposed rats in anxiety tests. However, the ratio of open arms time in the plus maze was reduced by half in SMF-exposed rats. In the Morris water maze, SMF-exposed rats were partially impaired during the initial learning task as well as in the retention task at one week. We conclude that static magnetic field exposure altered emotional behaviors in the plus maze and led to cognitive impairments, or at least to substantial attention disorders, in the Morris water maze.     

Intra-Retrosplenial Cortical Grafts of Cholinergic Neurons: Functional Incorporation and Restoration of High Affinity Choline Uptake

Fetal septal neurons transplanted into the deafferented retrosplenial cortex (RSC) of rats have been shown to reinnervate the host brain and ameliorate spatial memory deficits. In the present study we examined the effects of implanting cholinergic neurons on high affinity choline uptake (HACU) in the denervated RSC and the correlational relationship between this cholinergic parameter and the level of behavioral recovery. Three groups of animals were used: 1) normal control rats (NC), 2) rats with lesions of the fornix and cingulate pathways (FX), and 3) lesioned rats with fetal septal grafts in the RSC (RSCsep-TPL). We found that intra-RSC septal grafts produced significant increases in HACU, and that recovery of HACU was significantly correlated with the improvements in the performance of spatial reference memory, spatial navigation, and spatial working memory tasks. We have also investigated the ability of the host brain to modulate the activity of the implanted neurons. In particular we evaluated the effect of the animals' performance in a 6-arm radial maze task on high affinity choline uptake (HACU). Animals in each of the NC, FX, and RSCsep-TPL groups were randomly assigned one of the following subgroups: 1) rats that performed the maze task before the determination of HACU (BEH), or 2) rats that did not perform the maze task before the determination of HACU (NON-BEH). Significant increases were observed in the NC and RSCsep-TPL groups, but not in the FX animals, indicating that fetal septal grafts in the RSC can become functionally incorporated with the host neural circuitry, and that the activity of the implanted cholinergic neurons can be modulated by the host brain.     

Lateral hypothalamic lesions and activity of the sympathetic nervous system

The firing rate of efferent sympathetic nerves to brown adipose tissue was measured on 18 h or 18 d following lateral hypothalamic lesions (LH). Eighteen hours following acute lateral hypothalamic lesions, sympathetic firing rate was significantly increased. Following chronic LH lesions there was a decrease in food intake and a fall in body weight which had stabilized by four days. Eleven days after surgery a group of control animals were food restricted and subsequently pair fed twice daily to maintain a body weight comparable to that of the LH lesioned animals. Food intake was lower in the pair-gained animals on all but one day of the experiment. When studied 18 days following LH lesions, sympathetic firing rates were significantly higher than in either the ad lib or pair-fed controls. Sympathetic firing rate in pair fed rats, on the other hand, was significantly lower than in the sham lesioned rats. These data are consistent with the hypothesis that the LH lesion removes an inhibitory control over sympathetic firing rate both acutely and in chronically lesioned animals and that this increased sympathetic firing rate may play an important role in the maintenance of a lower body weight.     

Cortical hypometabolism in injured brain: New correlations with the noradrenergic and serotonergic systems and with behavioral deficits

Changes with time after injury in behavioral deficits, as determined by the Morris swim test, and the in vivo specific binding of HEAT, a selective ₁-adrenoreceptor ligand, were compared with the time-course of development of cortical hypometabolism in rats with focal freezing lesions. In our trauma model, cortical hypometabolism was detectable in the lesioned hemisphere at 4 hr, became maximal (50% of normal) at 3 days and diminished towards normal on days 5 and 10 post-injury. Progressive impairment of acquisition of the Morris water maze task was demonstrated up to day 3 post-lesion with improvement thereafter. On day 3 the latency to reach criterion was 60% longer in lesioned animals than in corresponding sham-operated ones. An increase in the volume of distribution of HEAT, limited to cortical areas of the lesioned hemisphere, was demonstrable at 4 hr post-lesion and reached its maximum on day 3 (200% of normal) with subsequent return toward normal on days 5 and 10. Several types of drugs were shown previously to modify the cortical hypometabolism associated with cerebral injury. The present data indicate that the same drugs also modify the in vivo binding of HEAT and the behavioral deficits induced by brain lesions. Ibuprofen, a non-steroidal anti-inflammatory drug, p-chlorophenylalanine, an inhibitor of serotonin synthesis, ketanserin, a specific 5HT₂-receptor antagonist, and prazosin, an ₁-adrenergic receptor blocker all normalized the in vivo binding of HEAT in the cortical areas of the lesioned hemisphere. All groups of animals treated with these drugs also showed subtle, but statistically highly significant improvements in latency to locate the platform in the Morris water maze. Taken together these results show good correlation between behavioral deficits, changes in ₁-noradrenergic receptor binding and cortical hypometabolism in injured brain. This supports the hypothesis that post-injury cortical hypometabolism is a reflection of cortical functional depression in which both the serotonergic and noradrenergic neurotransmitter systems play a role, compatible with their inhibitory effects in the cortex and their postulated involvement in cortical information processing.Special issue dedicated to Dr. Leon S. Wolfe.