Neonatal alcohol exposure impairs acquisition of eyeblink conditioned responses during discrimination learning and reversal in weanling rats

Discrimination and reversal of the classically conditioned eyeblink response depends on cerebellar-brainstem interactions with the hippocampus. Neonatal "binge" exposure to alcohol at doses of 5 g/kg/day or more has been shown to impair single-cue eyeblink conditioning in both weanling and adult rats. The present study exposed neonatal rats to acute alcohol intubations across different developmental periods (postnatal day [PND] 4-9 or PND7-9) and tested them from PND26-31 on discriminative classical eyeblink conditioning and reversal. A high dose of alcohol (5 g/kg/day) dramatically impaired conditioning relative to controls when exposure occurred over PND4-9, but produced mild or no impairments when delivered over PND7-9. These findings support previous claims that developmental exposure period plays a critical role in determining the deleterious effects of alcohol on the developing brain. A lower dose of alcohol (4 g/kg/day) delivered from PND4-9--lower than has previously been shown to affect single-cue eyeblink conditioning--also produced deficits on the discrimination task, suggesting that discrimination learning and acquisition of responding to CS+ during reversal may be especially sensitive behavioral indicators of alcohol-induced brain damage in this rat model.     

The ontogeny of interstimulus interval (ISI) discrimination of the conditioned eyeblink response in rats

Discrimination of the eyeblink conditioned response (CR) between conditioned stimuli (CSs) of different durations and modalities was examined across development in rats. Interstimulus interval (ISI) discrimination was evident at Postnatal Days 23-34 in Experiment 1, and earlier CR peak latencies and enhanced CR amplitudes were seen to the long CS in the ISI discrimination group relative to a control group receiving the short CS without reinforcement. Experiment 2 showed that early CR peak latencies and enhanced CR amplitudes to the long CS in the ISI discrimination group were due to associative pairing of the short CS and unconditioned stimulus. Experiment 3 demonstrated ISI discrimination in adults that was improved relative to younger subjects, but with no enhancement of CR amplitude to the long CS in the ISI discrimination group. Cerebellar cortical maturation may influence the ontogeny of CR timing.     

A developmental comparison of trace and delay eyeblink conditioning in rats using matching interstimulus intervals

The effects of conditioning paradigm and interstimulus interval (ISI) were evaluated in this study of the development of associative learning in rats. The acquisition of classical eyeblink conditioning (EBC) was examined in two paradigms (trace vs. delay), with three different ISIs (short, medium, or long) at two ages (postnatal Days 21-23 or 29-31). These data provide the first parametric analysis of ISI in developing animals trained with trace EBC procedures. Further, by comparing trace and delay EBC, it was determined that when ISI is held constant, acquisition is similar for both conditioning paradigms regardless of developmental age. This suggests a similar ontogeny of associative processes for delay and trace EBC that relies on common neural substrates. However, conditioned response timing (onset and peak latencies) was affected by age and paradigm, suggesting that different neural mechanisms may play a role in timing delay versus trace conditioned responses.     

Binge-like ethanol exposure during the early postnatal period impairs eyeblink conditioning at short and long CS-US intervals in rats

Binge-like ethanol exposure on postnatal days (PD) 4-9 in rodents causes cerebellar cell loss and impaired acquisition of conditioned responses (CRs) during "short-delay" eyeblink classical conditioning (ECC), using optimal (280-350 ms) interstimulus intervals (ISIs). We extended those earlier findings by comparing acquisition of delay ECC under two different ISIs. From PD 4 to 9, rats were intubated with either 5.25 g/kg of ethanol (2/day), sham intubated, or were not intubated. They were then trained either as periadolescents (about PD 35) or as adults (>PD 90) with either the optimal short-delay (280-ms) ISI, a long-delay (880-ms) ISI, or explicitly unpaired CS and US presentations. Neonatal binge ethanol treatment significantly impaired acquisition of conditioning at both ages regardless of ISI, and deficits in the acquisition and expression of CRs were comparable across ISIs. These deficits are consistent with the previously documented ethanol-induced damage to the cerebellar-brainstem circuit essential for Pavlovian ECC.     

Effect of delay interval on classical eyeblink conditioning in 5-month-old human infants

Associative learning was evaluated in human infants with simple delay classical eyeblink conditioning. A tone conditioned stimulus (CS) was paired with an airpuff unconditioned stimulus (US) at three different delay intervals (250, 650, and 1,250 ms). Independent groups of healthy, full-term 5-month-old human infants were assigned to these three paired conditions and received two identical training sessions 1 week apart. The two longer delays resulted in associative conditioning, as confirmed by comparison with unpaired control groups. However, only at the 650-ms delay were associative eyeblinks adaptively timed to avoid the airpuff. The delay function at 5 months of age appears much sharper than is observed in adults. Together with the findings of A. H. Little, L. P. Lipsitt, and C. Rovee-Collier (1984), the present study suggests a downward shift in the optimal delay interval for associative eyeblink conditioning between 1 and 6 months of age. However, this delay remains longer than what is typically reported in adults.     

Long-term retention of the classically conditioned eyeblink response in rats

Retention of the classically conditioned eyeblink response in rats was tested with a conditioned stimulus (CS)-alone extinction test and 2 sessions of reacquisition training. Retention of the eyeblink conditioned response (CR) during both tests was highest 24 hr and 1 month after initial acquisition. Three months after initial acquisition, responding during the CS-alone test was at baseline, but there was significant savings during reacquisition. By 6 months after initial acquisition, the memory for the eyeblink CR was not expressed in either test. The group differences in retention, despite initial acquisition of the eyeblink CR to equal levels, suggest that rat eyeblink conditioning may provide a useful behavioral model for studying the neural processes underlying memory retention and loss.     

Effects of ipsilateral cerebellum ablation on acquisition and retention of classically conditioned eyeblink responses in rats

The ipsilateral cerebellum to the trained eye has been reported to be essential for acquisition and retention of the conditioned response (CR) in rabbit eyeblink conditioning. Although pharmacological studies have suggested its important roles in other species too, to what degree does eyeblink conditioning in rats depend on the ipsilateral cerebellum is not clear. In this work, we ablated the ipsilateral cerebellum in rats before or after conditioning to examine its roles in acquisition and retention of the CR. In the first experiment, rats received ablation of the ipsilateral cerebellum and recovered for more than 3 weeks. They then underwent eyeblink conditioning for 7 days with a tone and a periorbital electrical shock. Consistent with other previous reports, hemicerebellectomized rats showed significant impairment compared to sham-lesioned rats. However, the hemicerebellectomized rats acquired CRs to some degree, and the acquired CR showed adaptive timing. In the second experiment, rats received the hemicerebellectomy after acquiring CR by 7 days of conditioning in a delay paradigm. After more than 3 weeks of recovery, they were again conditioned in a delay paradigm. Rats with ipsilateral cerebellar lesions showed severe impairment in retention of the pre-acquired CR; however, they reacquired CR to some degree during the subsequent reconditioning sessions. These results suggest that the ipsilateral cerebellum plays an important role in rat eyeblink conditioning as well but that other brain regions can partially compensate for its removal.     

Connections of the caudal anterior cingulate cortex in rabbit: neural circuitry participating in the acquisition of trace eyeblink conditioning

The caudal anterior cingulate cortex (cAC) is an essential component of the circuitry involved in acquisition of forebrain-dependent trace eyeblink conditioning. Lesions of the cAC prevent trace eyeblink conditioning [Weible AP, McEchron MD, Disterhoft JF (2000) Cortical involvement in acquisition and extinction of trace eyeblink conditioning. Behav Neurosci 114(6):1058-1067]. The patterns of activation of cAC neurons recorded in vivo suggest an attentional role for this structure early in training [Weible AP, Weiss C, Disterhoft JF (2003) Activity profiles of single neurons in caudal anterior cingulate cortex during trace eyeblink conditioning in the rabbit. J Neurophysiol 90(2):599-612]. The goal of the present study was to identify connections of the portion of the rabbit cAC previously demonstrated to be involved in trace eyeblink conditioning, using the neuronal tract tracer wheat germ agglutinin conjugated to horseradish peroxidase, to better understand how the cAC contributes to the process of associative learning. Reciprocal connections with the claustrum provide a route for the transfer of sensory information between the cAC and neocortical and allocortical regions also involved in learning. Connections with components of the basal forebrain cholinergic system are described, with relevance to the proposed attentional role of the cAC. Reciprocal and unidirectional connections were in evidence in multiple thalamic regions, including the medial dorsal nucleus, which have been implicated in a variety of conditioning paradigms. Anterograde connections with the caudate and lateral pontine nuclei provide access to forebrain motor and brainstem sensory circuitry, respectively. The relevance of these connections to acquisition of the trace conditioned reflex is discussed.     

Temporal discrimination in the cerebellar cortex during conditioned eyelid responses

Little is known about mechanisms used by the nervous system to encode time. In light of recent evidence, cerebellar cortex involvement in the learned timing of conditioned eyelid responses shows promise as an area of investigation into neural timing mechanisms. Lesion studies indicate that the cerebellar cortex is necessary for response timing, but do not rule out the possibility that response timing is encoded afferent to the cerebellum. To differentiate between precerebellar and cerebellar cortical timing mechanisms, rabbits were trained by pairing direct stimulation of mossy fibers in the cerebellum as the conditioned stimulus (CS) with an eyeshock unconditioned stimulus (US). We find that individual animals can produce diffently timed conditioned responses when trained with a mossy fiber CS that has been paired with the US at various interstimulus intervals. The fact that differently timed responses can be conditioned using constant-frequency stimulation of an invariant subset of mossy fibers as the CS suggests that timing information in the afferent input to the cerebellum is not essential. Two rabbits trained with single-pulse stimulation in the cerebellum as the CS also learned differently timed conditioned responses; suggesting that fiber recruitment during a stimulus train does not convey the necessary temporal coding to the cerebellar cortex. Together with the lesion data, these findings suggest that the learned timing of conditioned eyelid responses occurs in the cerebellar cortex. Received: 25 August 1997 / Accepted: 12 January 1998     

Medial dorsal thalamic lesions impair blocking and latent inhibition of the conditioned eyeblink response in rats

The effects of lesions of the medial dorsal thalamic nucleus (MD) on blocking and latent inhibition (LI) of the rat eyeblink response were examined in the present study. Previous work has demonstrated that the cingulate cortex and related thalamic areas are involved in processing conditioning stimuli throughout training. The experiments in the present study tested the hypothesis that disruption of cingulothalamic stimulus processing produced by lesions of the MD would impair 2 types of associative learning that involve decremental changes in attention. In Experiment 1, MD lesions severely impaired blocking. In Experiment 2, MD lesions severely impaired LI. The results indicate that lesions of the MD impair incremental, decremental, or both types of changes in stimulus processing during learning.     

Effects of sectioning the corpus callosum on interocular transfer in hooded rats

Summary After learning a vertical/horizontal visual discrimination with one eye rats were tested on the same or the reverse discrimination with the second eye. Positive but imperfect savings were found in transfer rats and zero savings in reversal rats. However, there were no differences between the savings scores of normal rats and rats with section of the corpus callosum. When the first eye was subsequently retested, the normal rats which had learned the reverse discrimination with the second eye were significantly worse than those tested on transfer with the second eye, whereas no such difference occurred in the operated groups. It is argued that the uncrossed optic pathway is responsible for the indistinguishable performance of normal and operated rats on second eye performance, but that performance when the first eye is retested is now influenced by callosal fibres connecting primary traces in each hemisphere.     

Single-unit activity in red nucleus during the classically conditioned rabbit nictitating membrane response

Previous investigations have suggested that the cerebellum and associated brainstem structures, including the red nucleus, are essential for the expression of the classically conditioned nictitating membrane (NM) response. The present study examined the firing patterns of extracellularly-recorded single units in the red nucleus of the awake rabbit during differential conditioning. Tones were used as conditioned stimulus (CS+ and CS−) and periocular electrostimulation was used as the unconditioned stimulus (US). Most units exhibited one or more changes in firing rate during the presentation of the CS, and increases in firing were much more common than decreases. The onset of some of these changes appeared to be time-locked to the onset of the CS (‘CS-locked’ responses), while other changes were time-locked to the onset of the CR (‘CR-locked’ responses). About one-third of all CS-locked changes were CR-dependent, meaning that the neuronal response was reduced when the CR did not occur. About two-thirds of all CR-locked responses preceded the onset of the CR, and lead times varied considerably across units. Many CR-locked units were located in what has been described as a dorsal face region of the red nucleus. Most units responded to the US, and some of the US responses were CR-dependent: i.e., a smaller US response was evoked when a CR preceded the US than when the CR was absent. Our results support the notion that cerebellum-brainstem circuits are involved in generating NM CRs.     

Visual discrimination learning and interhemispheric transfer in the cat,as affected by 6-hydroxydopamine

Summary Learning and interhemispheric transfer of visual flux, pattern and form discriminations were studied in the cat after selected exposure of one suprasylvian cortex to 6-hydroxydopamine (6-OHDA). Biochemical assay using High Performance Liquid Chromatography (HPLC) two weeks after 6-OHDA revealed no discernible norepinephrine or dopamine in the treated cortex, but elevated concentrations of these transmitters in the cortex of the opposite hemisphere. Visual discriminations learned before treatment with 6-OHDA were retained at a high level using either the eye on the side of chemical lesion or the eye on the untreated side. An asymmetric deficit in learning new form discriminations was present, however, when the eye on the untreated side was used, in contrast to normal learning using the eye on the side of the hemisphere with depleted adrenergic nerve supply. Once learning was achieved using the lesioned hemisphere transfer of the engram was found to the untreated hemisphere. Thus, the unlesioned hemisphere was unable to learn normally using direct retinal input from the ipsilateral eye, but showed good capacity for learning using indirect visual input from the contralateral eye. This suggests a powerful influence of the callosum on the learning abilities of the two hemispheres, an influence proved by sectioning the callosum. Callosotomy resulted in a reversal of the discriminative capacities seen after 6-OHDA, i.e. the lesioned hemisphere was defective relative to the unlesioned hemisphere.This research was supported in part by USPHS research grant EY-00577, EY-01583 and SO7-RR05415     

Effects of the intertrial interval on taste-aversion learning in rats

Variations in the intertrial interval (ITI) between two taste-aversion conditioning trials indicated that more saccharin-aversion learning occurs with longer ITIs (Experiments 1, 2 and 4). Two conditioning trials separated by 35 min did not produce stronger taste aversions than a single saccharin-lithium pairing; however, stronger conditioning was evident when the two trials were administered with a 3-day intertrial interval. Independent evaluation of saccharin and lithium exposure as possible sources of proactive and retroactive interference suggested that poor conditioning at short ITIs occurs because lithium treatment during Trial 1 interferes with conditioning during Trial 2 (Experiment 2). Assessments of the unconditioned suppression of activity (Experiments 3) and novel-fluid intake (Experiment 4) indicated that drug treatment was effective during both conditioning trials regardless of the intertrial interval. Furthermore, with a 35-min ITI (but not with a 3-day ITI) drug treatment in Trial 1 persisted long enough to summate with the unconditioned responses to drug treatment in Trial 2. These findings are consistent with the idea that with short ITIs the lingering effects of the US from Trial 1 overshadow or mask the CS flavor in Trial 2, and this interferes with the conditioning that Trial 2 would otherwise produce.     

The effect of experience on the development of tactual-visual transfer in pigtailed macaque monkeys

The study described here is the first to experimentally demonstrate the effects of experience on the development of tactual-visual transfer. Infant pigtailed macaque monkeys (Macaca nemestrina) were reared from birth to 2 months of age in special cages that allowed the separation of tactual and visual experience. When assessed on a battery of measures at the end of the 2-month period, animals reared without the opportunity to integrate information across the two sensory modalities performed at chance levels on a paired-comparison measure of tactual-visual transfer and performed worse than controls in a visually guided reaching task. After living in the standard laboratory environment for 2 additional months, they were reassessed. While their visually guided reaching now no longer differed from that of controls, they continued to perform at chance on the tactual-visual transfer assessment and their performance on this task was significantly worse than the control groups. Performance on visual acuity and visual recognition memory measures did not differ between groups at either age, suggesting that the deficit was limited to tactual-visual functioning. The results are discussed in terms of a possible sensitive period during which specific environmental input is required for the development of normal tactual-visual cross-modal processing.     

Persistence of classical conditioned heart rate after extensive lesions of the striatum in rats

The participation of the Corpus Striatum in the acquisition and maintenance of conditioned heart rate was determined. After a 25 min period of adaptation in a sound attenuated chamber 8 rats were presented with a single association of a sound (CS) and a nociceptive stimulus (US) applied to the paws, while electrocardiographic recordings were made. Twenty-four hours later the sole application of CS produced a significant increment in the heart rate. The same results were obtained in 8 rats that were previously electrolytically lesioned in the corpus striatum. In another 8 rats the electrolytic lesions were performed after the acquisition session. These rats also showed the increment in heart rate produced by the CS during the testing session. Finally, in 8 more rats in which the association of CS-US was inversed no conditioned response was obtained. The electrolytic lesions comprised several milimeters of the corpus striatum involving the anterior medial and posterior part and even extended to surrounding areas. Nevertheless the animals were able to acquire and retain the vegetative conditioned response.     

Serial reversal learning of position discrimination in developing rats

The current study established a procedure to evaluate the capability of rats on postnatal days (PND) 21, 26, and 30 to perform a spatial serial reversal task using a T-maze. Training consisted of an acquisition session followed by a series of six reversal sessions. To examine the role of proactive interference in the serial reversal effect, the point of reversal was manipulated so that it occurred at the start of each session (between-sessions) or the midpoint of each session (within-sessions). Performance was initially impaired during the first reversal but improved dramatically across the series. Reversal between-sessions enhanced this serial reversal effect in comparison to reversal within-sessions. Experiment 1 showed that rats of all ages learned the between-sessions serial reversal task at a comparable rate. However, on the within-sessions task, PND21 rats were impaired relative to the PND26 and 30 rats, which did not differ. Experiment 2 revealed that the addition of a tactile cue that is correlated with each phase of reversal eliminated age and task differences in serial reversal performance. These findings suggest that higher-order cognitive processes underlying serial reversal are present during the weanling period, but there is some improvement with age under conditions involving high memory interference and/or difficulty in detecting the transition between reversal phases.     

Interocular transfer in albino rats as a function of forebrain or forebrain plus midbrain commissurotomy

An attempt was made to determine whether interocular transfer of a black-white discrimination would be blocked by section of interhemispheric pathways. Thirty-six albino rats were equally divided into three surgical treatment groups: sham-operate, corpus callosum-section, or corpus callosum-plus superior colliculus- and posterior commissure-section. Before statistical analyses of the behavioral data were performed, the animals were reassigned to appropriate groups based upon histological examination of the lesions. The multiple-commissurotomized animals showed nearly complete incidental section of hippocampal and habenular commissures, while the callosal-sectioned and sham-operated animals had comparatively little incidental damage. The callosal-sectioned and sham-operated animals exhibited interocular transfer of a black-white discrimination, although the callosal-sectioned animals showed less transfer than the sham operates. The multiple-commissurotomized animals showed no transfer; they also made reliably more errors during acquisition via the second eye (i.e., during the test for transfer) than either the sham operates or corpus callosal-sectioned animals. These results are comparable to previous research on cats. In addition, there were no differences in the initial rates of acquisition of the black-white discrimination between the three groups; these data suggest that section of the commissures does not impair the normal mechanisms needed to acquire the discrimination.     

Mother-pup interactions during a short olfactory deprivation period in young rats

The effects of short-term bilateral naris occlusion (inducing olfactory deprivation) on mother-pup interactions, suckling behavior and hormonal status during post-natal development in Wistar rats were studied. Bilateral naris occlusion was performed on 8-day-old rat pups and its effects were evaluated at Day 9 and at Day 15. The narins opened spontaneously between Day 12 and 14. Olfactory-deprived pups exhibited a greater level of corticosterone at both ages versus untreated or sham animals. Olfactory deprivation via naris occlusion, in young rats, alters mother-pup interactions with a decrease in the duration of mother-pup retrieving and an increase in pup licking. Olfactory-deprived pups showed also a lower mean duration of nursing and a decrease in nipple attachment, which appeared related to difficulties in finding the nipple. Olfactory-deprived pups had difficulty recognizing their nest. These behavioral alterations were accompanied by a diminution in milk ingested and growth retardation associated with a reduced level of thyroxin at both 9 and 15 days of age.