Memory impairments following basal forebrain lesions

The functional contribution of the nucleus basalis magnocellularis (NBM) and medial septal area (MSA) to memory was evaluated in 4 behavioral tasks. The tasks were postoperative acquisition of a win-stay spatial discrimination in a T-maze, a win-shift spatial discrimination on a radial arm maze, active avoidance in a shuttle box, and passive avoidance in a shuttle box. Bilateral lesions were made by injecting ibotenic acid (IBO) into the NBM or MSA. Control rats received operations in which no neurotoxin was injected. When compared to controls, rats with lesions in either the NBM or MSA had significantly impaired choice accuracy in the T-maze and radial maze tasks, took significantly fewer trials to reach criterion in the acquisition, but not the retention of an active avoidance task, and significantly more trials to reach criterion in the passive avoidance task. The results show that equivalent behavioral changes are obtained from lesions in the NBM and MSA in tasks that vary in their type of motivation, reinforcement, response-reinforcement contingency, and response. These behavioral changes suggest that the NBM and MSA may both be involved in memory.     

Selective lesions of basal forebrain cholinergic neurons produce anterograde and retrograde deficits in a social transmission of food preference task in rats

We examined the performance of Long-Evans rats with 192 IgG-saporin lesions of the medial septum/vertical limb of the diagonal band (MS/VDB) or nucleus basalis magnocellularis/substantia innominata (NBM/SI), which removed cholinergic projections mainly to hippocampus or neocortex, respectively. We studied the effects of these lesions on anterograde and retrograde memory for a natural form of hippocampal-dependent associative memory, the social transmission of food preference. In a study of anterograde memory, MS/VDB lesions did not affect the immediate, 24-h or 3-week retention of the task. In contrast, NBM/SI lesions severely impaired immediate and 24-h retention. In a study of retrograde memory in which rats acquired the food preference 5 days or 1 day before surgery and they were tested 10-11 days after surgery, MS/VDB-lesioned rats showed striking memory deficits for the preference acquired at a long delay (5 days) before surgery, although all lesioned rats exhibited poorer retention on both retest sessions than on their pretest performance. Subsequent testing of new anterograde learning in these rats revealed no disrupting effects of lesions on a standard two-choice test. When rats were administered a three-choice test, in which the target food was presented along with two more options, NBM/SI-lesioned rats were somewhat impaired on a 24-h retention test. These results provide evidence that NBM/SI and MS/VDB cholinergic neurons are differentially involved in a social memory task that uses olfactory cues, suggesting a role for these neurons in acquisition and consolidation/retrieval of nonspatial declarative memory.     

Transplantation of embryonic ventral forebrain grafts to the neocortex of rats with bilateral lesions of nucleus basalis magnocellularis ameliorates a lesion-induced deficit in spatial memory

Embryonic ventral forebrain grafts containing developing cholinergic cells were transplanted to the neocortex of rats with bilateral quisqualic acid lesions of the nucleus basalis magnocellularis. A lesion-induced deficit on performance of a spatial alternation test of memory was reduced by such transplants. When the same animals were treated with the acetylcholinesterase inhibitor physostigmine (0.05 mg/kg), however, performance on the behavioral task was not further promoted, and therefore, under these conditions, the cholinergic cortical transplants appear not to be subject to modulation by anticholinesterase drugs.     

Behavior alters the uptake of [3h]choline into acetylcholinergic neurons of the nucleus basalis magnocellularis and medial septal area

Behavioral experience changed sodium-dependent high affinity choline uptake (SDHACU) in the hippocampus and frontal cortex. Rats were trained on various behavioral tasks and sacrificed after testing. SDHACU was determined in frontal cortex and hippocampus, areas that receive cholinergic innervation from the nucleus basalis magnocellularis (NBM) and the medial septal area (MSA), respectively. Untrained rats taken directly from their home cages had fairly consistent levels of SDHACU in the hippocampus (1.76 ± 0.45, X ± S.E.) and frontal cortex (1.46 ± 0.37). In the hippocampus of rats performing in a radial maze and T-maze and in rats that surpassed a criterion level in an active avoidance task, SDHACU increased significantly above Cage (untrained) group levels. In the cortex of rats performing the radial maze task, SDHACU decreased slightly. There were no other changes in frontal cortical SDHACU. After behavioral testing ceased, SDHACU in rats performing the radial maze task remained elevated above Control and Treadmill group levels for 20 days, but returned to near control levels 40 days later. Our data demonstrate that a functional differentiation exists between the MSA and NBM cholinergic systems, and that the measurement of SDHACU in central cholinergic neurons is a useful tool to identify the influences of behavior and environment upon changes in neurochemical events and neuronal activity.     

Lesions of perirhinal cortex produce spatial memory deficits in the radial maze

Rats with bilateral electrolytic lesions of perirhinal cortex (PRC) or sham control (SHAM) lesions were tested in spatial reference and working memory tasks in the radial arm maze. In experiment 1, one arm of the maze was baited and always located in a fixed position relative to the extra-maze environment. PRC lesioned animals made a significantly greater number of errors than did SHAM animals during initial training in this reference memory task and exhibited a delay-dependent impairment on trial 5 in a series when a delay period of 5, 60, 120, or 240 s was inserted between trials 4 and 5. In experiment 2, when a second group of the animals was tested on the standard radial arm maze working memory task, the performance of the PRC group was markedly impaired relative to controls. These data demonstrate that electrolytic PRC lesions result in a deficit in both spatial reference and spatial working memory tasks. These effects are interpreted as being consistent with the idea that PRC plays an important role in episodic memory processes. These processes may include the storage of information, which is required for the performance of spatial tasks. Hippocampus 1998;8:114–121. © 1998 Wiley-Liss, Inc.     

The role of two major cholinergic systems in memory acquistion and retention in the eight-arm radial maze

The role of the nucleus basalis magnocellularis (NBM) and the medial septum (MS) in postoperative memory acquisition and postoperative memory retention was assessed in the eight-arm radial maze task (Olton). NBM lesions and MS lesion significantly decreased choline acetyltransferase (CAT) activity in the innervated areas. Animals with MS lesion, but not NBM lesions, showed significant impairment of memory acquisition. NBM lesions induced a unique strategic pattern, but the development of strategic pattern with training was comparable to the sham-injected group. Animals with MS lesion did not develop strategic pattern with training. NBM and MS lesions did not influence either memory retention or developed strategic pattern. These results suggest that (1) MS contributes to the radial maze task; (2) the two systems have qualitatively different contributions in the radial maze task; (3) the two systems do not contribute to memory retention; (4) the two major cholinergic systems have different functions in the pathophysiology of Alzheimer's disease.     

The effects of NMDA-induced retrohippocampal lesions on performance of four spatial memory tasks known to be sensitive to hippocampal damage in the rat

Four separate cohorts of rats were employed to examine the effects of cytotoxic retrohippocampal lesions in four spatial memory tasks which are known to be sensitive to direct hippocampal damage and/or fornix-fimbria lesions in the rat. Selective retrohippocampal lesions were made by means of multiple intracerebral infusions of NMDA centred on the entorhinal cortex bilaterally. Cell damage typically extended from the lateral entorhinal area to the distal ventral subiculum. Experiment 1 demonstrated that retrohippocampal lesions spared the acquisition of a reference memory task in the Morris water maze, in which the animals learned to escape from the water by swimming to a submerged platform in a fixed location. In the subsequent transfer test, when the escape platform was removed, rats with retrohippocampal lesions tended to spend less time searching in the appropriate quadrant compared to controls. Experiment 2 demonstrated that the lesions also spared the acquisition of a working memory version of the water maze task in which the location of the escape platform was varied between days. In experiment 3, both reference and working memory were assessed using an eight-arm radial maze in which the same four arms were constantly baited between trials. In the initial acquisition, reference memory but not working memory was affected by the lesions. During subsequent reversal learning in which previously baited arms were now no longer baited and vice versa, lesioned animals made significantly more reference memory errors as well as working memory errors. In experiment 4, spatial working memory was assessed in a delayed matching-to-position task conducted in a two-lever operant chamber. There was no evidence for any impairment in rats with retrohippocampal lesions in this task. The present study demonstrated that unlike direct hippocampal damage, retrohippocampal cell loss did not lead to a general impairment in spatial learning, implying that the integrity of the retrohippocampus and/or its interconnection with the hippocampal formation is not critical for normal hippocampal-dependent spatial learning and memory. This outcome is surprising for a number of current hippocampal theories, and suggests that other cortical as well as subcortical inputs to the hippocampus might be of more importance, and further raises the question regarding the functional significance of the retrohippocampal region. Introduction     

Impairment of semantic memory after basal forebrain and fornix lesion

Semantic memory impairment is classically associated with lesion of the anterior temporal lobe. We report the case of a patient with severe semantic knowledge impairment and anterograde amnesia after bilateral ischemic lesion of the fornix and of the basal forebrain following surgical clipping of an aneurysm of the anterior communicating artery. Fluorodeoxyglucose positron emission tomography (FDG-PET) showed a temporal hypometabolism. Severe semantic impairment is a rare complication after rupture of an anterior communicating artery aneurysm and may result from disconnection of the temporal lobe.     

Quisqualate lesions of rat NBM: Selective effects on working memory in a double Y-maze

Some authors have reported that quisqualic acid lesions of the nucleus basalis magnocellularis (NBM), although producing large cortical cholinergic losses, have little effect on memory. The purpose of the present study was to investigate the effects of quisqualic acid lesions of the NBM on working and reference memory in a double Y-maze. Each trial started with placement into one of the two end arms of the first Y-maze, and the correct response was to go down the stem (reference memory). Access was then given to the second Y-maze, the correct response being conditional upon the side of the first Y-maze from which that trial had begun (working memory). Rats were trained to an 88% correct criterion and were then given either bilateral quisqualic acid (60 nM, 0.5 microliters) or sham lesions (0.9% saline, 0.5 microliters) of the NBM. One week postsurgery, rats were tested on the double Y-maze task with delays of 0, 5 or 30 seconds being introduced prior to both the working and reference memory choice. NBM lesions produced a 63.2 +/- 6.2% decrease of cortical choline acetyltransferase (ChAT) compared to unoperated controls. Delays affected only the working memory of the sham group. Rats with lesions showed a significant impairment of working memory at all delays, but no change in reference memory. Results indicate that quisqualic acid lesions of the NBM that produce significant reductions in cortical ChAT selectively impair working memory.     

Impairment in T-maze reinforced alternation performance following nucleus basalis magnocellularis lesions in rats

Rats were trained on a reinforced alternation paradigm using an elevated T-maze. After pre-surgical training subjects received either ibotenic acid (4 micrograms/0.4 microliter) or vehicle (pH 7.4, 0.4 microliter) bilaterally into the region of the nucleus basalis magnocellularis--an important source of neocortical acetylcholine projections. Acetylcholinesterase staining of sectioned brains revealed a loss of neocortical, but not hippocampal staining in lesioned animals. On the T-maze task, lesioned rats showed significantly impaired choice performance relative to controls. They also demonstrated significant side biases, the degree of which was correlated with choice performance deficit.     

Mnemonic deficits in animals depend upon the degree of cholinergic deficit and task complexity

The role of cholinergic basal forebrain (CBF) neurons in mnemonic behaviors was investigated using the immunotoxin 192IgG-saporin. We assessed two routes of immunotoxin administration: intracerebroventricular (ICV) and intraparenchymal (INTRA). INTRA lesions of the medial septum (MS) and/or the nucleus basalis magnocellularis (NBM) were compared with ICV-lesions, INTRA-phosphate-buffered saline injected, and naive controls. The INTRA-NBM/MS and ICV NBM/MS lesions produced a similar depletion of choline acetyltransferase activity of 80% across all CBF projections. Water maze performance was similarly impaired for ICV- and INTRA-NBM/MS animals during various phases of testing, whereas animals with individual lesions of the NBM or MS performed at the level of controls. In contrast to the allocentric demands of water maze performance, the egocentric-based T-maze task revealed a vast group difference between the ICV- and the INTRA-NBM/MS animals. INTRA-NBM/MS animals showed a severe deficit in the non-match- and match-to-position version, whereas again, animals with single lesions were unimpaired. In addition, a dichotomy between animals with complete cholinergic deafferentation was observed in the inhibitory avoidance task. ICV-NBM/MS showed a diminished retention for the aversive stimulus while the INTRA-NBM/MS animals remembered well. During plus maze testing, only the INTRA-NBM/MS animals had a reduced level of anxiety. Although non-CBF regions may have been differently affected by the two routes of immunotoxin administration, global measures of arousal, motivation, and motor initiation did not reveal a different behavioral pattern. Our findings suggest that a dynamic interplay exists between the degree of cholinergic deficit and task demands revealing different types of mnemonic impairments.     

Neurochemical Effects Following AF64A Injection into the Basal Nuclei of Rats

Abstract: AF64A, a specific cholinergic neurotoxin, was injected into the basal nuclei of rats. The injected sites were the bilateral nucleus basalis of Meynert (NBM) and the medial septal nucleus (MSN), well known to be the nuclei of origin of the two major cholinergic pathways. The remote effects of injection were estimated by the regional choline acetyltransferase (CAT) activity in the frontal cortex, striatum and hippocampus. The injection of AF64A (1 nmol in 1 ul) produced a reduction in the CAT activity in each projected site: NBM lesions in the frontal cortex and MSN lesions in the hippocampus after one and three weeks. Twelve weeks after the injection, the reduced CAT activity had returned to normal levels. This neurochemical effect shows plasticity and recovery with time. The injections of small amounts of AF64A (0.2 and 0.1 nmol in 1 μl) produced no chemical changes after one week.     

Heterogeneity and selectivity of the degeneration of cholinergic neurons in the basal forebrain of patients with Alzheimer's disease

Cholinergic neurons were studied by immunohistochemistry, with an antiserum against choline acetyltransferase (ChAT), in the basal forebrain (Ch1 to Ch4) of four patients with Alzheimer's disease (AD) and four control subjects. ChAT-positive cell bodies were mapped and counted in Ch1 (medial septal nucleus), Ch2 (vertical nucleus of the diagonal band), Ch3 (horizontal nucleus of the diagonal band) and Ch4 (nucleus basalis of Meynert). Compared to controls, the number of cholinergic neurons in AD patients was reduced by 50% on average. The interindividual variations in cholinergic cell loss were high, neuronal loss ranging from moderate (27%) to severe (63%). Despite the small number of brains studied, a significant correlation was found between the cholinergic cell loss and the degree of intellectual impairment. To determine the selectivity of cholinergic neuronal loss in the basal forebrain of AD patients, NPY-immunoreactive neurons were also investigated. The number of NPY-positive cell bodies was the same in controls and AD patients. The results (1) confirm cholinergic neuron degeneration in the basal forebrain in AD and the relative sparing of these neurons in some patients, (2) indicate that degneration of cholinergic neurons in the basal forebrain contributes to intellectual decline, and (3) show that, in AD, such cholinergic cell loss is selective, since NPY-positive neurons are preserved in the basal forebrain.     

Lack of correlation between cortical levels of choline acetyltransferase and learning scores in rats with globus pallidus lesions

Previous work has shown that rats with lesions of the globus pallidus (GP) exhibit a generalized learning impairment. Data are presented suggesting that this impairment is not due to inadvertent damage to the nucleus basalis magnocellularis. Rats with GP lesions evidenced a significant visual discrimination learning loss and a significant reduction in cortical choline acetyltransferase (ChAT) activity. However, there was no significant correlation between the severity of the learning loss and the amount of reduction of cortical ChAT activity.     

Disruption of central cholinergic systems in the rat by basal forebrain lesions or atropine: effects on feeding, sensorimotor behaviour, locomotor activity and spatial navigation

Rats with ibotenic acid lesions of the nucleus basalis magnocellularis, the origin of the extrinsic cholinergic innervation of the cortex, were examined for changes in feeding, sensorimotor behaviour, nocturnal locomotor activity, and place navigation in the Morris swimming pool task, in comparison with control rats and rats receiving the muscarinic antagonist, atropine. The lesions produced acute feeding impairments, marked by weight loss and vigorous active rejection of food and water lasting 2-4 days, sensorimotor impairments in placing and orienting, and overnight hyperactivity. A similar hyperactivity was induced by atropine, lasting approximately 6 h following the injection. Rats with lesions or receiving atropine were similarly impaired in the acquisition of the spatial navigation task, they failed to reach control levels of efficiency even once they had acquired the task, and they showed small but significant retention impairments when pretrained in the absence of either treatment. The results are discussed in terms of the lesions producing a disruption of cortical cholinergic systems, with implications for the clinical disorder of senile dementia of the Alzheimer type, and in terms of possible associated disruption to non-cholinergic systems.     

Posttraining intrahippocampal infusion of a protein kinase AII inhibitor impairs spatial memory retention in rats

The role of protein kinase AII (PKA II) in spatial memory retention in male rats and its regulation of cholinergic gene expression were explored through the effects of intrahippocampal infusion of H-89, a selective PKA II inhibitor. Alterations in escape latency, travel distance, and swimming speed in a Morris water maze were measured. Animals were trained for 3 days; each day included two blocks, and each block contained four trials. Stereotaxic surgery was employed for the infusions after the last trial on the third day of training, and the animals were tested 48 hr after surgery. Bilateral intrahippocampal infusion of H-89 (2.5 or 5 microM) into the CA1 region generated significant alterations in escape latency and traveled distance but not swimming speed. The response was fairly dose dependent, and the maximal effect was obtained with 5 microM H-89. After behavioral testing, several of the infused animals were transcardially perfused and their brains removed. Brain tissue sections from these rats were subjected to immunohistochemical staining analysis with anticholine acetyltransferase (ChAT) antibodies. These analyses indicated that 5 microM H-89 infusions qualitatively reduced the density of ChAT-containing cholinergic nerve terminals in the dorsal hippocampus. The intrahippocampal infusions with 5 microM H-89 also caused an apparent reduction in the number of ChAT-containing neurons in the medial septum. Our results suggest that PKA II is involved in regulation of cholinergic gene expression and plays an important role in spatial memory retention in rats.     

Nucleus basalis lesions: Implication of basoamygdaloid cholinergic pathways in memory

Previous studies have shown a lack of association between cortical choline acetyltransferase (ChAT) activity and severity of memory impairment following excitotoxic lesions of the nucleus basalis magnocellularis (NBM). It recently has been proposed that the differential effects of NBM injections of various excitotoxins on amygdaloid and cortical ChAT may explain this result. The present study evaluated the mnemonic effect of unilateral intra-NBM infusions of the excitotoxins phthalic acid and quisqualic acid, which decrease ChAT activity primarily in the amygdala and cortex, respectively. Rats were trained in a double Y-maze, lesioned, and allowed to recover for 1 week prior to memory assessment. Behavioral results showed impaired working but not reference memory following phthalic acid lesions, and no significant effect following quisqualic acid lesions. Biochemical analysis in a second group of subjects confirmed that phthalic acid lesions produced a large decrease in basolateral amygdaloid ChAT, but had little effect on cortical ChAT activity. Conversely, quisqualic acid lesions produced a large decrease in cortical, but not basolateral amygdaloid, ChAT activity. These results suggest that the NOM amygdalopetal cholinergic pathways play a role in mnemonic functioning.     

Profound disturbances of spontaneous and learned behaviors following lesions of the nucleus basalis magnocellularis in the rat

It has been shown that a marked decline in the cortical activity of the cholinergic synthesizing enzyme choline-acetyltransferase (ChAT), accompanied by a severe neuronal loss in the nucleus basalis magnocellularis of Meynert occurs in the brains of patients with senile dementia of the Alzheimer type. However, the functional role of these neurons is largely unknown. In fact, very few studies have been done in animals. In this paper we report the behavioral effects of the lesion of the nucleus basalis magnocellularis in the rat either by radiofrequency current or by ibotenic acid injection at the level of the cell bodies. The two kinds of lesion lead to a profound disturbance of spontaneous and learned behaviors. There is a complete disorganization of behavior which is evidenced by an enhanced locomotor activity, an alteration in alimentary and hoarding behavior. In addition, we observed a deterioration of spatial memory and an incapacity to reverse a previously learned response. Biochemical assay showed that radiofrequency and ibotenic acid lesions produced a decrease of ChAT activity in the prefrontal and sensorimotor cortices and in amygdala without affecting the hippocampus or striatum. Ibotenic acid lesions seem to specifically destroy the cell bodies of the nucleus basalis magnocellularis since the dopaminergic and noradrenergic fibers of passage remained intact as measured by the unchanged level of endogenous catecholamine concentration in the terminal region in the prefrontal cortex. Presently, it cannot be said that the behavioral syndrome results solely from the lesion of the cholinergic neurons. Also, it is likely that the lesion of the nucleus basalis magnocellularis in the rat does not exactly reproduce the behavioral syndrome observed in Alzheimer's disease in man. However, this experimental approach in leading to a better knowledge of the functioning of these neurones could improve our understanding of this disease.     

Ventral hippocampal involvement in temporal order, but not recognition, memory for spatial information

The hippocampus is critical for spatial memory. Recently, subregional differences in the function of hippocampus have been described in a number of behavioral tasks. The present experiments assessed the effects of reversibly lesioning either the dorsal (dHip) or ventral hippocampus (vHip) on spontaneous tests of spatial recognition and temporal order memory. We report that although the dHip is necessary for spatial recognition memory (RM) (distinguishing a novel from a familiar spatial location), the vHip is involved in temporal order memory (the capacity to distinguish between two spatial locations visited at different points in time), but not RM. These findings and others are consistent with the hypothesis that temporal order memory is supported by an integrated circuit of limbic areas including the vHip and the medial prefrontal cortex.     

Mapping of the basal forebrain cholinergic system of the dog: A choline acetyltransferase immunohistochemical study

In an effort to produce a canine model of basal forebrain ischemia with memory deficits, we have shown that dogs possess a medial striate artery that perfuses basal forebrain territory, homologous to the human recurrent artery of Heubner. In the present study, we set out to delineate the precise topography of the cholinergic neurons in the canine forebrain, a neuronal system implicated in cognitive and memory functions. Floating coronal sections, derived from the head of the caudate nucleus to the rostral border of the hippocampus, were stained for choline acetyltransferase using a monoclonal antibody. Representative sections from one dog brain were drawn. These outlines were used for measurement of cell density, cell size, number of processes, and cell roundness. Choline acetyltransferase-positive neurons constituted four major subdivisions within the basal forebrain. A relatively dense population of cholinergic neurons was present in the medial septal nucleus (Ch1). A continuum of densely packed cells was also delineated within the vertical (Ch2) and horizontal (Ch3) nuclei of the diagonal band of Broca. A fourth group of heterogeneously packed cholinergic neurons represented the nucleus basalis magnocellularis (Ch4). Except for the caudal component of the Ch4 population, the forebrain cholinergic corticopetal system was located within the perfusion territory of the medial striate arteries. The Ch4 cell group in dogs is better defined than that of rodents but is not as sharply demarcated as in human and nonhuman primates. Our findings indicate that the dog may serve as an excellent model for assessing neurological and memory deficits, which, in humans, results from hypoperfusion of the recurrent artery of Heubner. © 1996 Wiley-Liss, Inc.