Ronald W. Skelton

The neuropharmacological and neurochemical basis of place learning in the Morris water maze

The Morris water maze (MWM) offers several advantages over other methods of studying the neurochemical basis of learning and memory, particularly with respect to its ability to dissociate deficits in memory formation from deficits in sensory, motor, motivational and retrieval processes. The contributions of nearly all of the major neurotransmitter systems have been investigated and consistent patterns have emerged. Normal function in glutamatergic and cholinergic systems is necessary for spatial learning, as blockade of NMDA receptors and cholinergic hypofunction prevents spatial learning but does not impair recall. Peptides such as adrenal and sex hormones and somatostatin may also be necessary for spatial learning. In contrast, activity in either GABAergic or opioidergic systems impairs spatial learning, though by quite different means. GABAergic activity prevents memory function, whereas opioidergic activity reduces motivation. Normal monoaminergic activity is necessary for normal performance in the MWM, but not for spatial learning per se. However, noradrenergic and serotonergic systems may enhance cholinergic-mediated mnemonic processes. Further research into the relative contributions of different receptor subtypes as well as interactions between neurochemical systems should provide significant advances in our understanding of the neural basis of learning and memory in mammals.

Bilateral cerebellar lesions disrupt conditioned eyelid responses in unrestrained rats.

Electromyographic eyelid responses in unrestrained rats were classically conditioned in a Pavlovian delay paradigm by using a tone conditioned stimulus (CS) and periorbital shock unconditioned stimulus (US). After eyelid conditioning was complete, bilateral electrolytic lesions were made in the dentate-interpositus region of the cerebellar nuclei. Initial eyelid conditioning was reliable and very similar to that previously observed in the rabbit, although the asymptotic eyelid responses contained a short-latency startle response in addition to the usual conditioned and unconditioned responses (CR and UR). Substantial decrements in CRs were observed in 13 of the 14 rats with accurately placed lesions. In contrast, startle responses and URs were unaffected. The results replicate the effects of cerebellar lesions on eyelid CRs in the rabbit and suggest that the anatomical basis of eyelid conditioning in both species is similar.

Eyeblink conditioning in the developing rat.

Long-Evans rat pups, 17-18 or 24 days of age, were trained with an eyeblink conditioning (EBC) procedure that was used previously with adult rats (Skelton, 1988). Pups received 3 sessions of delay conditioning in a single day at about 4-hr intervals (100 trials/session). Trials involved pairings of an auditory conditioned stimulus (2.8-kHz, 82-dB tone) and a periocular-shock unconditioned stimulus (US; 100 ms, 2 mA), which were presented 280 ms apart. EBC was observed at both ages, but older pups learned much more rapidly. Subsequent experiments established that this effect is associative (Experiment 2), that age differences in EBC cannot be attributed to differences in ability to respond or in sensitivity to the US (Experiment 3), and that EBC rate can be modulated by motivational state (Experiment 4). This preparation may help elucidate the relation between neural development and the ontogeny of learning.

Diazepam impairs acquisition but not performance in the Morris water maze.

Diazepam is known to produce anterograde amnesia in both humans and animals. The present investigation sought to determine if this impairment is a direct result of diazepams interference with mnemonic processes or a result of deficits in performance or retrieval. Diazepam (3 mg/kg) was administered prior to training in the Morris water maze either before or after the rats had acquired the location of a submerged escape platform. Diazepam was found to impair acquisition but not retrieval of spatial information and this impairment was not due to the sedative, hypothermic or state-dependent learning effects of diazepam. These results replicate previous findings in the Morris water maze and provide new evidence that this deficit is primarily mnemonic in nature.

Baclofen, a selective GABAB receptor agonist, dose-dependently impairs spatial learning in rats.

The present investigation assessed the effects of the selective GABAB receptor agonist baclofen (1, 3, and 6 mg/kg) on spatial learning in the Morris water maze, an aversively motivated spatial learning task. Potential anxiolytic and sedative effects of baclofen were also assessed in an open field. Baclofen dose-dependently reduced locomotion in the open field but had little effect on thigmotaxia (anxiety). In the water maze, baclofen dose-dependently impaired spatial learning and reduced swim speed. During the probe trial given after training, only rats treated with the highest dose of baclofen (6 mg/kg) failed to show a bias for the correct quadrant. Following four additional retraining trials, a second drug-reversal probe trial was given and it was found that rats switched from saline to the highest dose of baclofen (6 mg/kg) showed a bias for the correct quadrant, as did rats switched from the two lowest doses of baclofen (1 and 3 mg/kg) to saline. Rats switched from the highest dose of baclofen (6 mg/kg) to saline failed to show a quadrant bias. Performance on a visible platform task was not impaired by baclofen at any dose. Together these results suggest that baclofen resembles GABAA agonists/positive modulators in that it impairs spatial learning, but not performance of a previously acquired escape response; but differs in that it does not reduce thigmotaxia (anxiety). Potential mechanisms by which baclofen impairs mnemonic processes are discussed.

Dorsal noradrenergic bundle lesions fail to disrupt self-stimulation from the region of locus coeruleus.

Intracranial self-stimulation (ICSS) from the region of the locus coeruleus (LC) was not attenuated by ipsilateral dorsal noradrenergic bundle (DNB) lesions. Bilateral DNB lesions also failed to affect LC ICSS, whereas the same lesions, in the same animals, resulted in a marked enhancement of lateral hypothalamic (LHA) ICSS responding. Behavioral, neuroanatomical and fluorescent histochemical data confirmed destruction of the DNB, thus suggesting that the dorsal noradrenergic system is not necessary for ICSS from the region of the LC.

Kindling‐Related Changes in Afterdischarge “Thresholds”

Amygdaloid stimulations were applied to rats at gradually increasing current intensities once every 60 sec until an afterdischarge (AD) was recorded through the stimulation electrode. These initial AD “thresholds” (x = 75 μA) were reduced over a 4‐day period by a series of 12 subthreshold stimulations. In contrast, stimulations administered on the same regimen but at a higher intensity (400 μA) had raised the AD threshold. The decreases in the AD threshold were relatively permanent, whereas the elevated thresholds were subsequently reduced to control levels by a series of subthreshold stimulations or by a 7‐day stimulation‐free period. Two days following the completion of Experiment 1, all of the rats were injected with a low dose of pentylenetetrazol. The incidence of pentylenetetrazol‐induced epileptic symptoms was much higher in the experimental animals than in the control subjects previously subjected only to the threshold estimation procedure.

Virtual environment navigation tasks and the assessment of cognitive deficits in individuals with brain injury.

Navigation in real environments is often impaired by traumatic brain injury (TBI). These deficits in wayfinding appear to be due to disruption of cognitive processes underlying navigation and may in turn be due to damage to the hippocampus and frontal lobes. These wayfinding problems after TBI were investigated using a virtual simulation of a Morris Water Maze (MWM), a standard test of hippocampal function in laboratory animals. The virtual environment consisted of a large virtual arena in a very large virtual room whose walls provided views of a naturalistic landscape. Eleven community-dwelling TBI survivors and 12 comparison participants, matched for gender, age and education were tested to see if they could find a location in the arena marked by one of the following: (a) a visible platform, (b) a single proximal object, (c) a single proximal object among seven other distracter objects, or (d) distal features inside and outside the room. The proximal objects allowed participants to use egocentric (body-centered) navigational strategies that rely on relatively simple stimulus-response associations. The absence of proximal cues forced the participants to rely on distal features of the environment (room walls, landscape elements) and tested their ability to use allocentric (world-based) navigational strategies requiring cognitive mapping. Results indicated that the navigation of TBI survivors was not impaired when the proximal cues were present but was impaired when proximal cues were absent. These results provide more evidence that the navigational deficit after TBI is due to an inability to form, remember or use cognitive maps.

Spatial Deficits in a Virtual Water Maze in Amnesic Participants with Hippocampal Damage

The Morris water maze is a standard paradigm for the testing of hippocampal function in laboratory animals. Virtual versions of the Morris water maze are now available and can be used to assess spatial learning and memory ability in both healthy and brain injured participants. To evaluate the importance of the hippocampus in spatial learning and memory, we tested five amnesic participants with selective hippocampal damage using a virtual water maze called the Arena Maze. The amnesic participants with hippocampal damage were impaired on the invisible platform (place) task that required them to use distal cues, but were able to navigate almost as well as comparison participants when the invisible platform was marked by a single proximal cue. These results not only confirm that the hippocampus plays a necessary role in human navigation in large‐scale environments but also provides a new link between the mnemonic and navigational roles of the hippocampus.

PHARMACOLOGICAL DISSOCIATION BETWEEN THE SPATIAL LEARNING DEFICITS PRODUCED BY MORPHINE AND DIAZEPAM

This study sought to determine whether the place learning deficits produced by diazepam are a secondary result of opioid release. Rats pretreated with diazepam (3 mg/kg) or morphine (15 mg/kg) were trained in the Morris water maze. Diazepam impaired place learning-slowing acquisition and preventing the formation of a quadrant preference. Morphine also slowed acquisition, but did not prevent place learning, and impaired escape to a visible platform. Flumazenil blocked the deficits produced by diazepam, but not morphine. Naloxone (2 mg/kg) blocked the deficits produced by morphine, but not diazepam. A high dose of naloxone (10 mg/kg) slowed acquisition, and exacerbated the deficit produced by diazepam. These results demonstrate that diazepam interferes with mnemonic processes through endogenous benzodiazepine receptors, independently of opioidergic systems. Further, they suggest that morphine interferes with motivational processes through opioidergic systems, independently of endogenous benzodiazepine systems.