Etan J. Markus

Interactions between location and task affect the spatial and directional firing of hippocampal neurons.

When rats forage for randomly dispersed food in a high walled cylinder the firing of their hippocampal “place” cells exhibits little dependence on the direction faced by the rat. On radial arm mazes and similar tasks, place cells are strongly directionally selective within their fields. These tasks differ in several respects, including the visual environment, configuration of the traversable space, motor behavior (e.g., linear and angular velocities), and behavioral context (e.g., presence of specific, consistent goal locations within the environment). The contributions of these factors to spatial and directional tuning of hippocampal neurons was systematically examined in rats performing several tasks in either an enriched or a sparse visual environment, and on different apparati. Place fields were more spatially and directionally selective on a radial maze than on an open, circular platform, regardless of the visual environment. On the platform, fields were more directional when the rat searched for food at fixed locations, in a stereotypic and directed manner, than when the food was scattered randomly. Thus, it seems that place fields are more directional when the animal is planning or following a route between points of special significance. This might be related to the spatial focus of the rats attention (e.g., a particular reference point). Changing the behavioral task was also accompanied by a change in firing location in about one-third of the cells. Thus, hippocampal neuronal activity appears to encode a complex interaction between locations, their significance and the behaviors the rat is called upon to execute.

Sex differences and correlations in a virtual Morris water task, a virtual radial arm maze, and mental rotation

Different tasks are often used to assess spatial memory in humans compared to nonhumans. In order to bridge this paradigmatic gap, we used a within-subject design to test 61 undergraduates on three spatial memory tasks. One of these tasks, the Vanderberg 3D mental rotation task, is classically used to assess spatial memory in humans. The other two tests are virtual analogues of two tasks used classically to assess spatial memory in rodents: the Morris water task and an eight-arm radial maze. We find that males perform significantly better than females on the mental rotation task and in finding a hidden platform in the virtual Morris water task. Moreover, during a probe trial, males spend significantly more distance of their swim in the training quadrant, but males and females do not differ in navigating to a visible platform. However, for the virtual eight-arm radial maze, there is no sex difference in working memory errors, reference memory errors, or distance to find the rewards. Surprisingly, an examination of the correlations among the three tasks indicates that only mental rotation ability and Morris water task probe trial performance correlate significantly among the three tasks (i.e. there are no significant correlations with traditional measures the tasks, e.g. time or distance to completion). Hence, the Morris water task and the eight-arm radial maze do not assess spatial memory in the same manner, and even after equating factors such as motivation, stress, and motor demands, there still are procedural demands of the tasks that reinforce differential strategy selection during spatial memory. This suggests that caution should be taken when utilizing these two tasks interchangeable as tests of spatial memory.

Sex differences and estrous cycle changes in hippocampus-dependent fear conditioning

Male and naturally cycling female rats were tested in a fear conditioning paradigm that encompassed both hippocampus-dependent and -independent components. The females were both conditioned and tested for retention at the same stage of the estrous cycle, during either estrus or proestrus. Male rats followed a regime similar to that for the female rats. Approximately 2 weeks after conditioning, the animals were examined for retention of the spatial context and of an explicitly paired conditioning tone. All animals showed a similar degree of conditioning to the tone. However, female proestrous rats showed less spatial-contextual conditioning than did male or estrous female rats. These results suggest that the changes found during the proestrous part of the cycle are related to hippocampal information processing and not to general changes in learning ability, to shock sensitivity, or to state-dependent learning. The results are discussed and related to previous findings regarding estrous cycle changes in behavior, anatomy, and physiology.

Neocortical synaptogenesis, aging, and behavior: Lifespan development in the motor-sensory system of the rat

Little evidence presently exists on the development and aging of synaptic contacts and their relationship to behavior, particularly in nonvisual brain areas. To investigate this interrelationship, rats at a series of developmental ages [postnatal day 1 (P1) to P90] were initially examined on a battery of motor tasks. The battery, ranging from simple reflexive tests to tests of complex locomotor capacities, consisted of tactile-induced forelimb placing, chin-induced placing, body righting, climbing an inclined plane, traversing a narrow beam, and keeping up with a revolving wheel. Following completion of the behavioral testing, the animals, together with an additional group of aged (28- to 29-month-old) rats, were killed and their motor-sensory cortex was removed, stained with osmium tetroxide or ethanol phosphotungstic acid (EPTA), and examined under electron microscopy for density of synaptic contacts. Simple motor abilities such as tactile-induced placing was present by the end of the first postnatal week, with locomotor performance reaching a mature level by the end of the third postnatal week, and intermediate task abilities maturing within this range. Paralleling the development of complex locomotor skills was a sharp increase in synaptic density in the molecular layer of the motor-sensory cortex, commencing in the second postnatal week and peaking at P30. After P30 there was a sharp decline in synaptic density as well as a decline in performance on some motor tasks, although these two functions seemed to be occurring independently. There was a continued, but less dramatic synaptic loss evident in the aged rats.

Age-related deficits on the radial maze and in fear conditioning: Hippocampal processing and consolidation

Young adult, middle‐aged, and old male F‐344 rats were assessed for their hippocampal ability. This was accomplished by examining the animals on two different paradigms, each incorporating a simultaneous measure of hippocampal‐dependent and ‐independent processing. The animals were fear conditioned and then tested for retention of the conditioning context and tone. This was followed by an 8‐arm radial maze task which combined spatial working and cued reference memory elements. The two paradigms are compared in terms of task demands, potential confounds, and validity for aging studies. The results indicate that the performance of the animals on the two tasks is correlated. Age‐related deficits limited to the hippocampal aspects of the above tasks were found, with no deficits found in the analogous but hippocampus‐independent aspects of these tasks.

Potential anxiogenic effects of cannabinoid CB1 receptor antagonists/inverse agonists in rats: Comparisons between AM4113, AM251, and the benzodiazepine inverse agonist FG-7142

Cannabinoid CB1 inverse agonists suppress food-motivated behaviors, but may also induce psychiatric effects such as depression and anxiety. To evaluate behaviors potentially related to anxiety, the present experiments assessed the CB1 inverse agonist AM251 (2.0-8.0mg/kg), the CB1 antagonist AM4113 (3.0-12.0mg/kg), and the benzodiazepine inverse agonist FG-7142 (10.0-20.0mg/kg), using the open field test and the elevated plus maze. Although all three drugs affected open field behavior, these effects were largely due to actions on locomotion. In the elevated plus maze, FG-7142 and AM251 both produced anxiogenic effects. FG-7142 and AM251 also significantly increased c-Fos activity in the amygdala and nucleus accumbens shell. In contrast, AM4113 failed to affect performance in the plus maze, and did not induce c-Fos immunoreactivity. The weak effects of AM4113 are consistent with biochemical data showing that AM4113 induces little or no intrinsic cellular activity. This research may lead to the development of novel appetite suppressants with reduced anxiogenic effects.

Hippocampal dysfunction during aging I: Deficits in memory consolidation☆

Numerous ablation studies indicate a critical role for the hippocampal system in establishing or consolidating certain types of memory. Normal aging manifests by selective neurobiological changes in the hippocampal formation and on performance of tasks that require a functional hippocampus, including retention of contextual fear conditioning. To determine if impairments in the consolidation process contribute to memory dysfunction in aging, middle-aged and aged rats were fear conditioned and subsequently received dorsal hippocampal lesions or sham surgery after a 1, 7, 14, or 28-day interval. During retention tests, middle-aged rats exhibited a temporally graded retrograde amnesia of contextual fear conditioning, whereas aged rats manifested contextual memory impairments at all intervals. We postulate that the lack of consolidation in aged animals relates to previous findings of age-related changes in neuroanatomy and neurophysiological plasticity. The present findings suggest that impaired hippocampal consolidation contributes to age-related learning and memory deficits.

Synaptic structural changes during development and aging.

Although a great deal is known about the development of synaptic number, comparatively little is known about the effects of development, and particularly aging, on the structure of the synapse. The present study examined synaptic structure in the molecular layer of the motor-sensory neocortex during early development (postnatal days (P) 1, 3, 5, 7, 10, 15, 20, 30), adulthood (P60, 90), and old age (28 months). Tissue was stained with osmium tetroxide (osmium) or ethanol phosphotungstic acid and the following synaptic characteristics were quantified: (1) presynaptic element length, area, thickness, maximal projection height and smoothness, and number and size of vesicles adjacent to the presynaptic element; (2) postsynaptic element length, area, and thickness; and (3) cleft width. There is an early developmental increase in synaptic element length, followed by an increase in thickness into adulthood. During development the height and width of the presynaptic dense projections increase, after which they remain stable. While the number of adjacent synaptic vesicles increases throughout the lifespan, there is a parallel decrease in their size. During the period of rapid synaptogenesis in this brain region there are no decreases in any of the synaptic structural parameters examined, indicating that newly generated synapses are either formed the same size as the existing mature synapses, or are extremely plastic and grow very rapidly. Unlike age-associated changes in synaptic number, no changes were found in synaptic structure during aging.

Effects of mild food deprivation on the estrous cycle of rats

It has long been known that severe food deprivation disrupts the estrous cycle. One of the main problems with behavioral tasks that use food for reinforcement is the requirement that the animal be food deprived. This manipulation could be problematic in studies using female animals, since it may interfere with the estrous cycle of the animals. The purpose of the present study was to investigate: (1) the effect of mild food deprivation on four different strains of rats, (2) factors in the food deprivation procedure that could affect the estrous cycle, and (3) the possible effect of enriched diets during food deprivation on the estrous cycle. A comparison of the estrous cycle in four different rat strains revealed differences in the reliability of the estrous cycle even before the onset of food deprivation. Fischer, Long-Evans, and Sprague-Dawley rats all showed reliable cycle patterns. This was not the case for Brown Norway rats. During food deprivation, the cycle of the Fischer rats was disrupted, whereas the Long-Evans and Sprague-Dawley animals continued to cycle. Both the rate of weight loss and the percent of ad libitum body weight were related to cessation of the estrous cycle. However, enriching an animals diet with sugar or oil additives delayed the disruption of the estrous cycle. Additionally, animals resumed cycling when returned to ad libitum weight levels. The present findings suggest that when animals need to be food deprived, preference should be given to using Long-Evans or Sprague-Dawley rats. If Fischer rats must be used, they should not be deprived below 90-95% of their ad libitum body weight. Strategies for future food deprivation studies are discussed, as well as a comparison of the effects of mild and severe food deprivation.

Age‐related deficits in the ability to encode contextual change: A place cell analysis

Aging is known to impair the formation of episodic memory, a process dependent upon the integrity of the hippocampal region. To investigate this issue, hippocampal place cells were recorded from middle‐aged and old F‐344 male rats while running on a “figure‐8” track. The top and bottom arcs of the track were removed, converting it into a plus maze, and the animals were required to conduct a working memory task. Following this change in task, the arcs were replaced and the animals again ran the figure‐8 task. Analysis of place fields across the recording session demonstrated that both middle‐aged and old rats had reliable representations of the figure‐8 task. A comparison of place fields between different behavioral tasks (figure‐8 and plus maze) demonstrated a change in the hippocampal representation of the environment in both age groups, despite the fact that the animals remained on the maze throughout the recording session. Notably, place cells in old animals were less affected by the change in task than those in middle‐aged animals. The results suggest that hippocampal neurons reflect significant behavioral events within a given environment. Furthermore, the data indicate that age‐related episodic memory deficits may result from decreased sensitivity of the hippocampal network to respond to meaningful changes in the environment. Hippocampus 10:338–350, 2000