Pamela S. Hunt

Impaired trace fear conditioning following neonatal ethanol : Reversal by choline

Neonatal ethanol exposure in animals results in performance deficits on tests of hippocampus-dependent spatial memory, and recent studies have shown that extra dietary choline can ameliorate some of these impairments. In this experiment, rats were administered 5.25 g/kg ig ethanol per day or sham intubations on Postnatal Days (PD) 4-9 and choline (0.1 ml of an 18.8 mg/ml solution) or saline subcutaneously on PD 4-20. On PD 30, rats were given delay or trace fear conditioning trials and were tested for conditioned stimulus-elicited freezing 24 hr later. Neonatal ethanol produced a profound impairment in trace conditioning that was reversed by choline. Groups did not differ in delay conditioned responding, indicating that neonatal ethanol produces a relatively selective cognitive deficit that can be alleviated with supplemental choline.

Deficits in trace fear conditioning in a rat model of fetal alcohol exposure: dose-response and timing effects.

In humans, prenatal alcohol exposure can result in significant impairments in several types of learning and memory, including declarative and spatial memory. Animal models have been useful for confirming that many of the observed effects are the result of alcohol exposure, and not secondary to poor maternal nutrition or adverse home environments. Wagner and Hunt (2006) reported that rats exposed to ethanol during the neonatal period (postnatal days [PDs] 4-9) exhibited impaired trace fear conditioning when trained as adolescents, but were unaffected in delay fear conditioning. The present series of three experiments represent a more detailed analysis of ethanol-induced deficits in trace conditioning. In Experiment 1, the dose of ethanol given to neonates was varied (3.0, 4.0, or 5.0g/kg/day). There was a dose-dependent reduction in trace conditioning, with the poorest performance observed in animals treated with the highest dose. In Experiment 2, it was found that the impairment in trace conditioning resulting from neonatal ethanol exposure was dependent on the duration of the trace interval used for training; less learning was evident in ethanol-exposed animals trained with longer trace interval durations. These results confirm other reports of delay-dependent memory deficits. Finally, Experiment 3 determined that ethanol exposure limited to the first half of the neonatal period (PDs 4-6) was more detrimental to later trace conditioning than exposure during the second half (PDs 7-9). These results support the hypothesis that trace-conditioning impairments resulting from early ethanol exposure are due to the drugs teratogenic effects on the developing hippocampus, as the findings parallel those observed in animals with discrete hippocampal lesions. Comparisons between delay and trace fear-conditioning performance in animals exposed to ethanol during the brain growth spurt provide a model system to study both selective learning impairments and possible treatment approaches for humans with fetal alcohol spectrum disorders.

Delayed development of fear-potentiated startle in rats.

The developmental emergence of fear-potentiated startle was examined in rats ranging in age from 16 to 75 days. In Experiment 1, a pure tone served as the conditioned stimulus (CS) and an acoustic startle pulse served as the unconditioned stimulus (US) for fear conditioning. Fear-potentiated startle by the tone CS was observed in rats 23 days of age and older but not in rats 16 days of age. In Experiment 2, a light served as the CS. Rats 30 days of age and older showed fear-potentiated startle, whereas 23-day-old rats did not. The final experiment demonstrated that another behavioral index of fear, stimulus-elicited freezing, was observed earlier in development than fear-potentiated startle, confirming the effectiveness of the training procedure for conditioning fear. The results suggest that fear-potentiated startle is a relatively late-emerging response system, paralleling the development of conditioned autonomic changes (e.g., heart rate) rather than that of freezing.

Trace and long-delay fear conditioning in the developing rat

In two experiments with rats, we examined the developmental emergence of conditioned freezing following trace and short-delay conditioning and also included a long-delay comparison group. In the short-delay and trace groups, a 10-sec conditioned stimulus (CS) was paired with shock; for the trace rats, a 10-sec trace interval followed CS termination. The long-delay groups received a 20-sec CS paired with shock, to equate for the longer interstimulus interval (ISI) in the trace group. Trace conditioning emerged later in development than did short-delay conditioning (see Moye & Rudy, 1987). Importantly, long-delay conditioning emerged in parallel with trace conditioning, at a similar time, and with similar strength. These findings suggest a role for the longer ISI, as opposed to the unfilled gap per se, in the late emergence of trace conditioning. The role of the hippocampus in trace conditioning and the possibility that young rats encode the temporal relationship between CSs and unconditioned stimuli are also considered.

Autonomic and behavioral correlates of appetitive conditioning in rats

The cardiac responses accompanying conditioned stimulus- (CS)-generated (orienting) and unconditioned stimulus- (US)-generated appetitively motivated behaviors (P. C. Holland, 1977) were investigated. On the basis of contemporary psychophysiological research, CS-generated responses were predicted to produce bradycardia, and US-generated responses to produce tachycardia. Pairing a 10-s visual CS with food delivery produced conditioned behavioral orienting (rearing) during the initial portion of the CS, followed by magazine approach (US-generated) responses as the CS progressed. CS onset produced a decrease in heart rate, mediated by an increase in parasympathetic stimulation of the heart, which persisted throughout the 10-s CS; no support for a biphasic cardiac response was observed. These data are discussed with respect to other conditioned autonomic responses and their relevance to foraging and food ingestion.

Emergence of conditioned cardiac responses to an olfactory CS paired with an acoustic startle UCS during development: Form and autonomic origins

Pairing an olfactory conditioned stimulus (CS) with an acoustic startle unconditioned stimulus (UCS) produced conditioned bradycardia in rats ranging from 12 days of age to adulthood but failed to produce conditioning in 10-day-olds (Experiment 1). The second experiment compared the effects of two different UCSs, acoustic startle and electric shock. The startle UCS produced conditioned bradycardia accompanied by immobility. In contrast, the shock UCS produced conditioned tachycardia accompanied by behavioral activation. Pharmacological analysis revealed that activation of the parasympathetic nervous system (PNS) produced the conditioned bradycardia (Experiment 3) and that activation of the sympathetic nervous system (SNS) produced the conditioned tachycardia (Hunt, Hess, & Campbell, 1994). Further comparisons revealed that the startle UCS established conditioning several days earlier in development than the shock UCS. The ability of the PNS to respond to phasic stimulation earlier in development than the SNS was discussed as a possible mechanism for the early development of conditioned bradycardia.

Post-training ethanol disrupts trace conditioned fear in rats: Effects of timing of ethanol, dose and trace interval duration

Ethanol has complex effects on memory performance, although hippocampus-dependent memory may be especially vulnerable to disruption by acute ethanol intoxication occurring during or shortly after a training episode. In the present experiments, the effects of post-training ethanol on delay and trace fear conditioning were examined in adolescent rats. In Experiment 1, 30-day-old Sprague-Dawley rats were given delay or trace conditioning trials in which a 10s flashing light CS was paired with a 0.5 mA shock US. For trace groups, the trace interval was 10 s. On days 31-33, animals were administered ethanol once daily (0.0 or 2.5 g/kg via intragastric intubation), and on day 34 animals were tested for CS-elicited freezing. Results showed that post-training ethanol affected the expression of trace, but had no effect on delay conditioned fear. Experiment 2 revealed that this effect was dose-dependent; doses lower than 2.5 g/kg were without effect. Experiment 3 evaluated whether proximity of ethanol to the time of training or testing was critical. Results show that ethanol administration beginning 24h after training was more detrimental to trace conditioned freezing than administration that was delayed by 48 h. Finally, in Experiment 4 animals were trained with one of three different trace intervals: 1, 3 or 10s. Results indicate that post-training administration of 2.5 g/kg ethanol disrupted trace conditioned fear in subjects trained with a 10s, but not with a 1 or 3s, trace interval. Collectively the results suggest that ethanol administration impairs post-acquisition memory processing of hippocampus-dependent trace fear conditioning.

Pharmacological dissociation of trace and long-delay fear conditioning in young rats

In most studies comparing trace and delay conditioning, CS duration is kept constant across training conditions but the interstimulus interval (ISI), the time from CS onset to US onset, is confounded. In the infrequently used long-delay condition, however, ISI is kept constant across the trace and delay conditions but CS duration varies. A recent study reported that trace and long-delay fear conditioning have the same developmental trajectory, with both emerging later in development than standard-delay conditioning (). Past studies have shown that trace conditioning is mediated by the cholinergic system; given the parallel developmental emergence of trace and long-delay conditioning, the present study examined whether the cholinergic system also mediates long-delay conditioning. Two experiments, both involving Sprague-Dawley-derived rats and using freezing as a measure of learned fear, showed that the cholinergic system is critically involved in trace conditioning but is not involved in long-delay conditioning. Specifically, pre-training injections of the muscarinic receptor antagonist scopolamine impaired acquisition of a CS-US association in 32-day-old rats trained with a trace procedure but had no effect on rats this age trained with a long-delay procedure (Experiment 1). Similarly, pre-training injections of physostigmine, a cholinesterase inhibitor, enhanced acquisition of trace conditioning in 25-day-old rats but had no effect on long-delay conditioning in rats this age (Experiment 2). Taken together, the results indicate that despite the similarities between trace and long-delay conditioning in terms of developmental emergence and level of conditioned responding, they are mediated by different physiological systems.

A further investigation of the developmental emergence of fear-potentiated startle in rats

This series of experiments was designed to reexamine the ontogenetic emergence of the fear-potentiated startle response in rats. Previous results (Hunt, Richardson, & Campbell, 1994) indicated that potentiated startle to a light conditioned stimulus (CS) paired with an acoustic unconditioned stimulus (US) was not observed until 30 days of age. In the present experiments, subjects were given pairings of a light CS with a brief footshock unconditioned stimulus (US) and were tested for fear-potentiated startle 24 hr later. Subjects 23 and 30 days of age exhibited significant potentiated responding in the presence of the light, while 17-and 20-day-olds did not. Subjects 17 days of age did reliably express conditioned decreases in heart rate to the light at the 24-hr test. The failure to observe fear-potentiated startle at the youngest age was shown not to be due to a general disruption of conditioned fear responding by either (a) pretest startle stimulus presentations or (b) contextual characteristics of the startle testing apparatus. The capacity to express fear through a potentiated startle response develops later than the capacity for other defensive responses in the rat.

Postnatal binge ethanol exposure affects habituation of the cardiac orienting response to an olfactory stimulus in preweanling rats

BACKGROUND Prenatal exposure to alcohol can result in intellectual impairments, reduced habituation and distractibility, visual-spatial deficits, and problems in attention. Dysfunction in attention, including habituation of responses to nonsignal stimuli, can have devastating consequences for cognitive development. This research examined whether binge exposure to ethanol on postnatal days (PD) 4 to 9 would affect cardiac orienting or response habituation to an olfactory stimulus in preweanling rats. METHODS Ethanol-exposed subjects were given ethanol 5.25 g/kg/day on PD4 to 9. Controls were either sham intubated or unhandled during this time. To assess baseline and phasic cardiac function, PD16 subjects were tested under differential pharmacological blockade of the sympathetic or parasympathetic nervous systems by administration of the peripherally acting antagonists atenolol or atropine methyl nitrate. In a second experiment, separate groups of subjects were tested for habituation of the cardiac orienting response to an olfactory stimulus. RESULTS Results indicate that postnatal ethanol had no observable effect on the functional development of autonomic control over heart rate. Similarly, ethanol exposure had no effect on the form or magnitude of the cardiac orienting response. However, neonatal ethanol exposure did result in a deficit in orienting response habituation; ethanol-exposed subjects continued to respond to the stimulus with a large-magnitude bradycardia after control subjects exhibited complete response habituation. In addition, ethanol-exposed subjects had longer orienting response latencies than controls. CONCLUSIONS The results indicate that this animal model has the potential to contribute to investigations designed to understand basic forms of memory impairments observed in humans with a history of prenatal alcohol exposure. The postnatal binge ethanol model of fetal alcohol effects produces deficits in nonassociative memory that are similar to those observed in human infants exposed prenatally to ethanol. Deficits in response habituation have important implications for cognitive development.