Conditioning of aversion to an odor paired with peripheral shock in the developing rat

Four experiments examined an apparent inability to associate, or severe deficiency in associating, an odor and a footshock during the first 2 weeks of life in the rat, a cue-to-consequence relationship that had formerly seemed age-dependent. With a particular classical conditioning procedure, however, significiant conditioning occurred on postnatal Days 6 and 10 with relatively few conditioning trials; the procedure employed an odor explicitly unpaired with footshock (CS?), as well as an odor paired with footshock (CS+) (Experiment I). Experiment II assessed the contribution of a CS? exposure in the conditioning of rats 8, 15, or 50 days of age. For 8-day-olds, exposure to both the CS+ and CS? resulted in conditioned aversion to the CS+ after eight but not one conditioning trials, but neither 1 nor eight trials with only a CS+ produced conditioning. For 15- and 50-day-olds, conditioning to the CS+ odor was significant after one trial with, but not without, a specific CS? with eight trials, however, conditioning was significant with or without the specific CS?. It was verified with the 50-day-olds in Experiment III that aversion to the CS+ was conditioned with a single trial only if a CS? had been presented, with a slight trend toward superior conditioning if the CS? preceded rather than followed the CS+ during conditioning. Experiment IV tested the hypothesis that exposure to the distinctive CS? odor sensitzes the animal to the specific properties of the CS+ odor. Fifteen and 50-day-old rats were given one conditioning trial with a CS+ odor that was either unaccompanied by a CS? or that was presented with a CS? odor in the same context as the CS+ or in a different context. For both 15? and 50-day-old rats, conditioning to the CS+ occurred only for animals given the CS? in the same context as the CS+, indicating that the hypothesis should be rejected. The results generally indicate rapid and substantial odor-footshock conditioning in rats as young as 6 days of age, with CS-exposure established as perhaps especially significant for conditioning early in life, but important for rats of all ages tested, from infancy to adulthood.     

Latent Inhibition and Conditioning in Rat Strains Which Show Differential Prepulse Inhibition

Latent inhibition (LI) is the retardation of associative conditioning resulting from preexposure of the conditioned stimulus (CS) alone prior to conditioning. Schizophrenic patients show deficient prepulse inhibition (PPI) and, at least acutely, deficient LI as well. We recently found that Brown Norway (BN) rats show a PPI deficit compared to Wistar-Kyoto (WKY) rats. If PPI and LI depend on neural processes with common genetic substrates, then LI should be deficient in BN rats as well. Here, LI of a conditioned taste aversion was examined in BN and WKY rats. One group from each strain was preexposed to a saccharin-flavored solution (CS) the day prior to conditioning. For taste aversion conditioning, these two groups again consumed saccharin and were injected with lithium chloride (unconditioned stimulus) 10 min later. A second group from each strain was not preexposed to the CS and was treated identically during conditioning, while a third group was not conditioned (injected with sodium chloride). To test for taste aversion conditioning, saccharin was offered for 20 min/day for 3 days. Nonconditioned BN and WKY rats consumed equal amounts of saccharin on test days. In both strains, conditioned rats showed a saccharin aversion. However, conditioning was less robust in BN than in WKY rats. WKY rats showed good LI of the conditioned taste aversion in that preexposed WKY rats consumed significantly more saccharin on test days than conditioned, nonpreexposed WKY rats. Preexposed BN rats did not consume significantly more saccharin on test days than conditioned, nonpreexposed BN rats. The previously reported deficiency in PPI in the BN rats was confirmed here 1 week after the taste aversion experiment. These results suggest that BN rats show deficient LI as well as PPI and display poor associative learning, a trait also reported in schizophrenia.     

Involvement of the anterior insular gustatory neocortex in taste-potentiated odor aversion learning

When an odor conditioned stimulus (CS) precedes illness (unconditioned stimulus; UCS), rats acquire relatively weak odor aversions. Conversely, when a compound odor-taste (flavor) CS precedes illness, rats acquire robust aversions both to the odor and to the taste components of a compound flavor CS. Thus, tastes potentiate odor-illness aversions during toxiphobic conditioning. Such conditioning effects have been referred to as taste-potentiated odor aversion learning (POA). Previous neurobehavioral experiments have shown that the anterior insular gustatory neocortex contributes to conditioned taste aversion (CTA) learning. The present experiment examined the involvement of the anterior insular gustatory neocortex in CTA learning and POA learning. To that end, four distinct groups of rats received bilateral electrolytic lesion placements in the orbitofrontal neocortex, the "somatic" gustatory neocortex, the anterior insular gustatory neocortex or the posterior insular neocortex. Control animals received anesthesia only. Subgroups of animals thereafter received aversion conditioning using either an odor (almond) CS or a compound odor-taste (almond-saccharin) CS. Aversions to the almond odorant and/or saccharin tastant were evaluated during extinction. Results indicated that animals lacking orbitofrontal neocortex or posterior insular neocortex acquired normal CTAs and POAs. Animals lacking somatic gustatory neocortex exhibited impaired CTA learning, yet those animals showed normal POA learning. Lesions centered in the anterior insular neocortex impaired both CTA learning and POA learning. These results demonstrate that the insular gustatory neocortex is uniquely involved in the higher-order integration of odors, tastes and illness.     

Ontogenetic differences in the expression of conditioned visual aversions

We examined ontogenetic differences in the expression of conditioned visual aversions. Sprague-Dawley-derived rats, 16 or 21 days of age, were conditioned with either an element (brightness) or compound (brightness/odor) CS+ and tested for their aversion to the common element (brightness). Aversions to the brightness cue were assessed by either a traditional test of preference between the CS+ brightness and a contrasting brightness or by assessment of freezing in the presence of either brightness cue. The results indicated that strength of conditioning as well as the expression of overshadowing/potentiation was dependent on the age of the animal and on the technique used to assess conditioning.     

Association of stimuli at long intervals in conditioned odor aversion

Rats learn to avoid a tasteless odorized solution if they experience visceral malaise after consuming it. This phenomenon is referred as Conditioned Odor Aversion (COA). It is widely accepted that an odor can only be associated with illness if the inter-stimulus interval (ISI) is shorter than 15 min. However, this conclusion is based on long-term memory tests usually made 48 h after conditioning, thus precluding the possibility to discriminate between a specific failure to make the odor-malaise association rather than the failure to consolidate the short-term association into long-term memory. In the present study, we compared the short-term and long-term memories for COA in rats trained with long ISIs.Independent groups of male rats were conditioned using 5, 15, 30, 60 or 90 min ISIs and tested either 4 or 48 h after conditioning. We found a reliable odor aversion at 5, 15, 30 and 60 min, but not at 90 min ISIs, when tested 4 h after conditioning. In contrast, odor aversion was only found at 5 and 15 min ISIs in the groups tested 48 h after training. Our results show that COA can be acquired when malaise follows the odor CS by at least 60 min. This finding indicates that the lack of aversion at long ISIs is not due to an association failure, but rather to a limitation in consolidating short-term memory into long-term memory of COA.     

Impact of continuous versus intermittent CS-UCS pairing on human brain activation during Pavlovian fear conditioning

During Pavlovian fear conditioning a conditioned stimulus (CS) is repeatedly paired with an aversive unconditioned stimulus (UCS). In many studies the CS and UCS are paired on every trial, whereas in others the CS and UCS are paired intermittently. To better understand the influence of the CS-UCS pairing rate on brain activity, the experimenters presented continuously, intermittently, and non-paired CSs during fear conditioning. Amygdala, anterior cingulate, and fusiform gyrus activity increased linearly with the CS-UCS pairing rate. In contrast, insula and left dorsolateral prefrontal cortex responses were larger during intermittently paired CS presentations relative to continuously and non-paired CSs. These results demonstrate two distinct patterns of activity in disparate brain regions. Amygdala, anterior cingulate, and fusiform gyrus activity paralleled the CS-UCS pairing rate, whereas the insula and dorsolateral prefrontal cortex appeared to respond to the uncertainty inherent in intermittent CS-UCS pairing procedures. These findings may further clarify the role of these brain regions in Pavlovian fear conditioning.     

Cortical and subcortical components of the conditioned saccharin aversion

Cortical mechanisms of the conditioned saccharin aversion were studied in 190 rats by the functional ablation technique. Water deprived animals had 15-min access to water on the familiarization Days 1 and 2. On Day 3 the rats were offered saccharin (0.1%, CS) followed 30 min later by LiCl injection (0.14 M, 2% body weight, UCS). The resulting intoxication caused marked conditioned saccharin aversion leading to reduced saccharin ingestion (by 60–80%) on Day 4. Unilateral cortical spreading depression (CSD) elicited on Days 3 and/or 4 by application of 25% KCl on the same or on opposite hemispheres does not diminish conditioned saccharin aversion in comparison with intact animals. Bilateral CSD elicited after saccharin administration, 15 min before or 5 min after LiCl injection on Day 3, does not prevent the development of conditioned saccharin aversion. On the other hand, forced feeding of saccharin under bilateral CSD elicits conditioned saccharin aversion neither with 30 min nor with 5 min CS-UCS delays although significant conditioned saccharin aversion was observed under forced feeding conditions in normal rats. It is concluded that cerebral cortex is necessary for the initial short-term storage of the indifferent gustatory trace but that the association of this engram with aversive gastrointestinal stimuli can be accomplished without cortical participation.     

Exposure to magnetic resonance imaging does not produce taste aversion in rats

A taste aversion test was used to evaluate possible toxic effects of magnetic resonance imaging (MRI). Thirty male Sprague-Dawley rats were randomly assigned to four groups: Group One (n = 10) received 30 minutes exposure inside the MRI scanner; Group Two (n = 10) received a sham exposure to the MRI scanner; Group Three (n = 5) was injected with 0.15 M lithium chloride; and Group Four (n = 5) was injected with vehicle. All groups were given 10 minutes access to a 0.1% saccharin solution immediately prior to their respective treatment. The rats treated with lithium chloride displayed a taste aversion to the saccharin solution upon subsequent testing over an eight day period. The two control groups (Two and Four) and the rats exposed to MRI did not display any aversion to the saccharin solution. These results are compared to other studies that have shown that magnetic fields can influence biological systems.     

Effect of odor quality and intensity on conditioned odor aversion learning in the rat

Odor quality and intensity were varied to test the ability of rats to associate odor with an induced illness. Rats were allowed 10 minutes access to water on each of nine days; deodorized air was directed towards each rat's nose while drinking at familiarization and recovery sessions (days 1-5 and 7-8, respectively) and odorized air at treatment and test sessions (days 6 and 9, respectively). Each rat was injected with LiCl following its drinking period on day 6. The difference between the amount of water consumed on day 6 and day 9 gave a measure of the conditioned aversion. Only mild or no aversion occurred with odors of n-butyric acid, benzylamine, cyclohexanone, and n-butanol. Strong conditioned aversions were obtained to odors of triethylamine, 1,4-cineole, and isoamyl acetate, and the degree of aversion increased linearly with the log of odor concentration. The effect of odor quality, intensity and presentation method, and the role of the different chemoreceptor systems in the acquisition of odor aversions are discussed.     

Basolateral amygdala NMDA receptors are selectively involved in the acquisition of taste-potentiated odor aversion in the rat

In the taste-potentiated odor aversion (TPOA) paradigm, animals acquire a strong aversion to an odor that is followed by delayed intoxication only if a gustatory stimulus is presented with the odor during conditioning. Although previous work has shown that N-methyl-D-aspartate (NMDA) receptors in the basolateral nucleus of the amygdala (BLA) play a role in the acquisition of TPOA, the present study aimed at describing the process in which NMDA receptors in the BLA are involved during acquisition of TPOA. Male Long-Evans rats received intra-BLA infusions of the competitive NMDA receptor antagonist D,L-2-2-amino-5-phosphonovalerate (D-APV; 0.05 and 0.50 microg) immediately before or after the odor-taste conditioned stimulus (CS) presentation, or immediately before the test. Results showed that D-APV impaired acquisition of TPOA when infused before, but not after, the CS presentation, but did not affect retrieval. These results suggest that NMDA receptors of the BLA are involved in the formation of potentiation--by taste--of the olfactory memory trace, but not in the maintenance of this process.     

When pentobarbital is the conditioned stimulus and amphetamine is the unconditioned stimulus, conditioning depends on the type of conditioned response

Injections of drugs into rats were used as conditioned stimuli (CSs) and as unconditioned stimuli (USs). With heart rate (HR) conditioning, the pentobarbital CS produces a higher HR than under control conditions. With avfail (aversion failure) conditioning, the pentobarbital CS loses much of its capacity to induce a conditioned taste aversion. HR conditioning was obtained with forward delays of up to 30 min and backward delays of up to 270 min, where the delays are defined by the interinjection interval. Avfail was obtained with forward delays of up to 270 min but not with backward delays. Neither HR conditioning nor avfail were context specific but could be demonstrated in a test apparatus after pairings that occurred in the home cage. This indicated that the external environment was not an important part of the effective stimulus complex. When HR conditioning was obtained, its latency and duration was not related to the delay between the CS and US injections or whether they were forward or backward.     

A Comparison of Two Model Systems of Associative Learning: Heart Rate and Eyeblink Conditioning in the Rabbit

In four classical conditioning experiments heart period and eyeblink responses were assessed concomitantly. The conditioned stimuli (CSs) were tones and the unconditioned stimulus (UCS) was a brief paraorbital electric shock. Using a 0.5-s duration CS, bradycardiac conditioned responses that consisted of a 4–5 ms change from pre-CS baseline occurred within 10–20 CS-UCS presentations. However, eyeblink conditioned responses began to occur only after 100–150 CS–UCS presentations. A 1.0-s CS duration resulted in bradycardiac conditioned responses of 15–30 ms change from pre-CS baseline, which again reached asymptote within 10–20 trials. Using a 4-s CS duration, in a differential classical conditioning paradigm, heart period discrimination between a reinforced CS+ and a nonreinforced CS? occurred within 10 trials; asymptotic performance of the heart period conditioned response occurred within 15 CS-UCS presentations and consisted of a bradycardiac response of 40–50 ms for the 12th interbeat interval following tone onset. These data thus indicate that these two model systems of mammalian learning (based on heart period and eyeblink responses) show quite different acquisition functions. It is also significant that heart rate slowing always accompanied the eyeblink conditioned responses, even though increases in general electromyographic activity as well as eyeblink conditioned responses were simultaneously observed during CS presentation.     

Disruption of conditioned taste aversion: the effect of ECS after the taste-illness interval

Rats were taught a conditioned taste aversion (CTA) by pairing a 10% sucrose solution (CS) with lithium chloride-induced poisoning (UCS) 30 min later. The extent to which electroconvulsive shock (ECS) (80 mA for 600 msec) impaired the acquisition of the CTA was studied by either interpolating the ECS within the CS-UCS interval or by administering it at various times following the UCS (0, 5, 10, 15, 30, 60 and 120 min). There was a pronounced although limited loss of learning among all groups when ECS was delivered no later than 10 min after the UCS. When ECS was administered between 15 and 120 min, the overall aversion acquired did not differ significantly from that of the poisoned controls. It is concluded that while the stability of CTA seems to increase at about the same time the behavioral concomitants of lithium toxicity become apparent (10 min), this phenomenon does not necessarily imply that the neural trace underlying CTA has consolidated into a less labile state.     

The development of olfactory conditioned ejaculatory preferences in the male rat. II. Parametric manipulation of conditioning session number and duration

We have previously demonstrated that repeated pairing of a neutral odor with copulation produces a subsequent conditioned ejaculatory preference (CEP) for females bearing that odor. The present study examines the course of CEP development. In Experiment 1, Long-Evans male rats were allowed access to almond-scented, sexually receptive females for either one, five, or nine conditioning sessions that were 30 min in duration. Males given five or nine sessions displayed significant CEPs. In Experiment 2, male rats were given a single conditioning session with multiple almond-scented females until either a duration (60, 120, 180, or 240 min) or copulatory criterion (two, four, or six ejaculatory series) was satisfied. Males that received 120-, 180-, or 240-min sessions or four ejaculations displayed significant CEPs; males that received two or six ejaculations displayed a trend for CEPs. Analysis of effect size estimates revealed that the strongest CEPs were produced by 120 min of copulation or four ejaculations. In Experiment 3, males receiving nine conditioning sessions each 30 min in duration displayed a more enduring CEP than did males receiving a single conditioning session 240 min in duration. These data suggest that early sexual experiences have particularly powerful influences on subsequent sexual preferences and that the development of sexual preferences are influenced by interactions between CS-UCS pairings and motivational variables.     

Acquisition of conditioned associations in Hermissenda: additive effects of contiguity and the forward interstimulus interval

Conditioned suppression of photokinesis by the marine mollusc Hermissenda was examined in 3 experiments. In each experiment, groups of animals received light (the conditioned stimulus, CS) that was paired with high-speed orbital rotation (the unconditioned stimulus, UCS), light and rotation explicitly unpaired, or no exposure to these stimuli. Twenty-four hours after training, all animals were tested for suppression of photokinesis in the presence of the light. To establish the effectiveness of our conditioning procedure, in Experiment 1 individual groups of animals received either 50, 100, or 150 CS-UCS pairings. Fifty pairings resulted in a marginal suppression of photokinesis, whereas 100 and 150 pairings produced strong suppression. In Experiment 2, the delay between CS onset and UCS onset was varied between 1 and 10 s. The 10-s interstimulus interval (ISI) did not support conditioning, whereas 1-s and 2-s ISIs were effective. As predicted by the current understanding of Hermissenda's neural network, in Experiment 3 it was found that CS-UCS pairings in which the CS preceded the onset of the UCS and terminated with the offset of the UCS evoked stronger conditioned suppression than either a CS that preceded the UCS and terminated with its onset or a CS that was paired in simultaneous compound with the UCS. This result indicates that CS-UCS contiguity as well as the forward ISI act additively to establish the CS-UCS association. In none of the 3 experiments were any differences observed between groups that were untreated and that received the CS and UCS unpaired. In total, these experiments suggest strong similarities in the temporal characteristics of associative learning in Hermissenda and vertebrate species.     

Backward conditioning of tumor necrosis factor-α in a single trial: changing intervals between exposures to lipopolysaccharide and saccharin taste

The current study examined the effect of backward conditioning with three different time intervals between exposures to lipopolysaccharide (LPS) as the unconditioned stimulus (US) and saccharin taste in water as the potential conditioned stimulus (CS). Forty-eight naïve female BALB/c mice at three months of age served as subjects, divided into six groups. Four groups were assigned to Experiment 1 for the tumor necrosis factor alpha (TNF-α) measure, and the remaining two groups were used in Experiment 2 to measure taste aversion behavior. Both experiments employed a single trial. The timing of introduction to the saccharin taste varied between 3 min, 7 h, and 24 h following an LPS injection in Experiment 1. Experiment 2 employed the three-minute interval only. These intervals correspond to distinct immunological, physiological, and behavioral events induced by LPS. On the day after re-exposure to the saccharin taste, the TNF-α groups were challenged with LPS to test the LPS tolerance response. While backward conditioning of taste aversion behavior was not observed, some evidence of conditioned TNF-α response and subsequent development of LPS tolerance was observed with backward conditioning in a single trial. This exploratory study demonstrated that the effect of backward conditioning on conditioned TNF-α response and LPS tolerance response in a single trial depended on the timing of when a CS is presented after LPS exposure.     

Estrogen-induced suppression of intake is not mediated by taste aversion in female rats

Estrogen treatment can suppress the intake of a previously presented gustatory conditioned stimulus (CS). This finding has been interpreted as an estrogen-induced conditioned taste aversion. However, a distinction must be made between taste aversion and taste avoidance. In particular, tastes are only considered aversive if they elicit a stereotypic behavioral response, otherwise the reduction in intake is classified as an avoidance. Although aversive orofacial responses have been reported in male rats after taste-estrogen pairings, they have not been examined in ovariectomized female rats. The goal of the present investigation, then, was to use similar procedures to determine whether conditioned aversion also mediates the estrogen-induced reduction of intake in female rats. Animals were introduced to a novel 0.1% saccharin solution and immediately thereafter were given a subcutaneous injection of vehicle or estradiol benzoate (10 microg). Responses were assessed using a two-bottle preference test, a one-bottle acceptance test, and a taste reactivity (TR) test. The results confirmed previous reports of a reduced preference for saccharin after saccharin-estradiol pairing using the two-bottle test. The reduction in intake during the one-bottle test, however, was not accompanied by stereotypic aversive responses, such as gaping. Surprisingly, a similar reduction in intake also occurred when using a backward conditioning procedure in which estrogen was injected before, rather than after, CS access. Thus, the present results show that the suppressive effects of estrogen reflect an avoidance, rather than aversion and, moreover, that the reduced intake may be due to an unconditioned, rather than a conditioned, response.     

Perceptual characteristics of the amiloride-suppressed sodium chloride taste response in the rat

The contribution of amiloride-sensitive membrane components to the perception of NaCl taste was assessed by using a conditioned taste aversion procedure. Eight independent groups of adult rats were conditioned to avoid either 0.1M NaCl, 0.5M NaCl; 0.1M NH4Cl, or 1.0M sucrose while their tongues were exposed either to water or to the sodium transport blocker amiloride hydrochloride. In contrast to rats exposed to water during conditioning, rats exposed to amiloride were unable to acquire a conditioned taste aversion to 0.1M NaCl. Differences in the acquisition of taste aversions between the amiloride- and nonamiloride-treated groups were not apparent when the conditioned stimulus (CS) was 0.5M NaCl, 0.1M NH4Cl, or 1.0M sucrose. Although the magnitude of the 0.5M NaCl aversion was similar between amiloride- and non-amiloride-treated rats, the perceptual characteristics of the CS differed between groups. Analyses of stimulus generalization gradients revealed that amiloride-treated rats generally avoided all monochloride salts after conditioning to 0.5M NaCl but not nonsodium salts or nonsalt stimuli. In contrast, rats not treated with amiloride only generalized the 0.5M NaCl aversion to sodium salts. No differences in generalization gradients occurred between groups when the CS was 0.1M NH4Cl or 1.0M sucrose. These findings suggest that the "salty" taste of NaCl is primarily related to the amiloride-sensitive portion of the functional taste response in rats. Conversely, the portion of the NaCl response insensitive to amiloride appears to have "sour-salty" perceptual characteristics and does not appear to be perceived as being salty.     

Conditioned food aversion elicited by the temperature of drinking water as a conditioned stimulus in rats

It is known that taste can act as a conditioned stimulus (CS) for conditioned food aversion. In the present study, in order to examine whether or not the temperature of drinking water can be a CS, we conducted behavioral experiments in Wistar rats. The following results were obtained: (1) The rats subjected to aversive conditioning to 5 or 40 degrees C distilled water could learn to avoid these CSs, but they did not avoid any taste stimuli. (2) The rats subjected to aversive conditioning to 5 or 40 degrees C 0.1 M sucrose developed a generalized avoidance to sucrose at any temperature. (3) When rats familiarized to 25 degrees C 5 mM saccharin-Na (Sacc) were subjected to aversive conditioning to 5 or 40 degrees C Sacc, they avoided the respective CS, but they did not generalize it to any other stimuli even if having the same temperature as the CS. (4) The rats which had undergone transection of the taste nerves (chorda tympani and glossopharyngeal nerves) could acquire the conditioned response to the temperature of the CS. These results suggest that rats can be conditioned to temperature aversion and that the taste nerves are not needed in the formation of this conditioning.     

Ontogenetic differences in the expression of odor-aversion learning in 4- and 8-day-old rats

Two experiments investigated the ability of preweanling rats, 4 or 8 days of age, to form an odor-LiCl association across various CS-US delays. The results of the odor preference test indicated that 8-day-old subjects acquired an aversion to the CS+ odor when trained with either a 0 or 15 min CS-US delay, while 4-day-old subjects did not exhibit a reduction in preference for the CS+ at any of the CS-US delays tested. The absence of a reduction in preference for the CS+ by 4-day-old rats could not be attributed to their failure to acquire the odor aversion, however; they avoided a texture with which the CS+ odor was paired in a second order conditioning paradigm. The results suggest that outcomes appearing to represent age-related differences in associative learning may in some instances be more appropriately viewed as representing ontogenetic differences in the way in which acquired associations are manifested in behavior.