Second-order conditioning of the rabbit's nictitating membrane response. Interstimulus interval and frequency of CS-CS pairings.

Second-order conditioning of the rabbit's nictitating membrane response (NMR) was investigated when second-order trials (CS1-CS2) were intermixed with first-order trials (CS2-US) from the outset of training. Experiment 1 showed that CR acquisition to CS1 was inversely related to the CS1-CS2 interval but nevertheless extended to an interval of 8,400 ms. Experiment 2 revealed that CR acquisition of CS1 was an inverted-U function of the number of CS1-CS2 trials relative to a fixed number of CS2-US trials. Experiment 3 directly contrasted second-order conditioning with a reinforced serial compound procedure (CS1-CS2-US) and a mixed procedure in which second-order trials were intermixed with the reinforced serial compound. Second-order conditioning was about half the strength of either the reinforced serial compound or the mixed procedure, which were similar. The present results are discussed with respect to the relative strength of excitatory and inhibitory processes in second-order conditioning.     

Second-order conditioning of the rabbit’s nictitating membrane response

Second-order conditioning of the rabbit’s nictitating membrane response (NMR) was investigated when second-order trials (CS1-CS2) were intermixed with first-order trials (CS2-US) from the outset of training. Experiment 1 showed that CR acquisition to CS1 was inversely related to the CS1-CS2 interval but nevertheless extended to an interval of 8,400 ms. Experiment 2 revealed that CR acquisition of CS1 was an inverted-U function of the number of CS1-CS2 trials relative to a fixed number of CS2-US trials. Experiment 3 directly contrasted second-order conditioning with a reinforced serial compound procedure (CS1-CS2-US) and a mixed procedure in which second-order trials were intermixed with the reinforced serial compound. Second-order conditioning was about half the strength of either the reinforced serial compound or the mixed procedure, which were similar. The present results are discussed with respect to the relative strength of excitatory and inhibitory processes in second-order conditioning.     

Cortisol and behavioral responses to separation in mother and infant guinea pigs

In Experiment 1, lactating guinea pigs 3 days postpartum exhibited an elevation of plasma levels of cortisol when their litters were taken from the home cage and placed out of auditory range for 30 min. If pups were left within auditory range during the period of separation, or if the litter was disturbed but not removed, no increase in plasma cortisol levels was observed. In Experiment 2, 11/12- and 18/19-day-old guinea pig pups placed alone in a novel environment for 30 min displayed levels of plasma cortisol and vocalizations that were greater than those of pups tested in the same environment but with their mother. Pups tested with littermates but not with their mother exhibited plasma cortisol levels that were as great as those of pups tested alone. Pups tested with littermates vocalized much less than pups tested alone but more than pups tested with their mother. These results indicate that brief mother-infant separation can activate the pituitary-adrenal system in mother as well as infant guinea pigs and they provide further evidence for the existence of a reciprocal mother-infant attachment in this species.     

Interactions between perinatal and neonatal associative learning defined by contiguous olfactory and tactile stimulation

Tactile stimulation of the neonate, as performed by the mother during and after delivery, has been described as an effective unconditioned stimulus during early ontogeny (Leon, 1987; Ronca & Alberts, 1994). The present experiments examined the interaction between perinatal and neonatal learning determined by the explicit association between alcohol odor and vigorous body stimulation of the perinatal organism. In Experiment 1, rat fetuses were exposed to either alcohol or saline 10 min prior to cesarean delivery. The alcohol administration procedure here employed was sufficient to provide sensory contamination of the amniotic fluid but avoid fetal alcohol intoxication. Pups in the two prenatal treatments later experienced the smell of alcohol, tactile stimulation, or both stimuli explicitly paired or unpaired. Other postnatal groups were composed of pups that had no explicit experience with either experimental stimulus. Pups subjected to alcohol odor in utero displayed more overall motor activity in response to that odor than saline controls. The increased motor responses were further potentiated in pups that experienced additional postnatal alcohol odor paired with tactile stimulation. In Experiment 2, pups were exposed to alcohol in the amniotic fluid 10 or 30 min prior to birth. As previously demonstrated the memory acquired in utero appears highly dependent upon contingency between exposure to this particular scent and delivery procedures. Pups in both prenatal treatment groups were then exposed to alcohol odor paired or unpaired with tactile stimulation. Some control animals received no further experience with either stimuli. Those pups exposed to alcohol odor paired with tactile stimulation both pre- and postnatally later showed maximum motor activity elicited by the odor of alcohol. The results support the notion of fetal associative learning comprising alcohol's chemosensory cues and behaviorally activating stimuli. Furthermore, the conditioned response under analysis is potentiated whenever neonates are reexposed to contingent presentations of the elements that defined the original associative memory.     

Responses of virgin mice, and of maternal females, to normal and caesarian-section delivered pups.

Pups were presented to adult mice of the Porton strain. Significantly more mothers and virgin females retrieved pups to the nest than did virgin males. More virgin females than males licked pups, built nests and assumed lactation posture. Differences were significant even if males that attacked pups were excluded from consideration. Latencies to first appearance of maternal activities were higher than reported by most other workers. Possible genetic strain differences in the response of virgin mice to pups are discussed. In Experiment 2 pups delivered by caesarian section with placentas attached were used. More mothers than either virgin males or females cleaned and accepted pups, indicating that pup cleaning may be controlled by factors different from those influencing other maternal activities.     

Conditioned aversions and their memories in 5-day-old rats during suckling

Association learning during suckling was investigated. Five-day-old rats equipped with tongue cannulae placed either 2 mm rostral or 4-6 mm caudal to the intermolar eminence received sweet or salty solutions while suckling. This ingestion was followed by lithium chloride toxicosis. Pups with anterior cannulae took in considerably less fluid than control pups when tested 5 or 16 days later. A series of control groups demonstrated that this acquired aversion was associative in nature. Pups with posterior cannulae did not form the association. The failure of 5-day-old rats with posterior cannulae to form associations while suckling is not due to the prevention of conditioning by the act of suckling per se. Rather, the failure rests in the fluid's not reaching anterior taste receptors when injected into the posterior oropharynx, where the nipple normally empties its contents. These findings are discussed in terms of the transfer of information obtained during suckling prior to weaning, to feeding and drinking during and after weaning.     

Odor-aversion learning and retention span in neonatal mouse pups

One hundred and sixty-four litters of Swiss CD-1 random-bred mice were used to assess learning and retention capacities during the first postnatal week. In Experiment 1, whole 7-day litters were exposed for 65 min to commercial extracts of either mint or lemon sprinkled over wood shavings. Five minutes after the beginning of the exposure, half of the litters were injected ip with the illness-inducing agent lithium chloride (LiCl; 0.20 M, 2% of body weight); the other half was treated with saline solution (8% NaCl). On Postnatal Day 10, the animals were singly introduced in a warmed arena for a 180-s preference test, and the time spent in the mint- and lemon-scented areas of the apparatus was recorded. When compared with saline-injected pups, mice that experienced lemon-LiCl pairings showed a significant aversion for the lemon-scented area, while the mint aversion in the mint-LiCl group just missed statistical significance. Three additional control groups (unhandled on Day 7, or only LiCl- or saline-injected) did not show significant preferences for either the mint or the lemon odor. In Experiment 2, litters of 3, 5, or 7 days were similarly exposed to lemon-scented shavings for either 5 or 20 min, injected with LiCl or saline, and then exposed for an additional 60 min to the shavings. On Postnatal Day 10, tests like those of Experiment 1 showed a significant odor-aversion in animals conditioned on Day 7, but not in those conditioned on Day 3 or 5. In Experiment 3, 3- and 5-day old pups were exposed to lemon odor-LiCl or -NaCl pairings, and tested for aversion after 3 or 7 days (CS duration 5 min before injection and either 30 or 60 min after injection). Only when the conditioning-testing interval was limited to 3 days did LiCl-injected groups show a significant aversion, which did not depend on duration of CS exposure.     

Olfactory stimulation induces filial preferences for huddling in rat pups.

Rat pups of all ages huddle with conspecifics, but the senosory control of contact behavior changes ontogenetically. Thermal cues control huddling until about Day 15, at which time species' odors become the dominant stimulus. The present experiments indicate that the filial response to conspecifics is dependent on olfactory experience. A synthetic chemical scent was added to the smells of the dam from Day 1 to Day 20 postpartum. Standardized videographic tests were used to assess the development of huddling preference. Preferences for nest-typical smells emerged by Day 15 in pups from both scented and nonscented litters. Pups from scented nests preferred to huddle with a scented stimulus rat, whereas control pups preferred a nonadulterated rat stimulus. Additional testing indicated that the affiliative preferences were specific to rearing odor and were not based on decreased aversion to test scents, or on disrupted olfactory discrimination. The ontogeny of species-typical contact behavior is discussed in terms of the induction of a perceptual preference that is based on early odor stimulation.     

Early (preweanling) recognition of alcohol's orosensory cues resulting from acute ethanol intoxication

Three experiments were conducted in order to analyze the possibility that during acute alcohol intoxication, preweanling rats process ethanol's orosensory cues. Intragastric administration of an ethanol dose equivalent to 3.0 g/kg resulted in peak blood alcohol levels greater than 250 mg% (Experiment 1). Twenty-four hours after receiving this dose, 11-day-old pups expressed an ethanol odor aversion in a locational test. This aversion response was not observed when pups were previously administered an ethanol dose resulting in blood alcohol levels lower than 150 mg% (Experiment 2). Nevertheless when subjects received such a lower dose and 30 to 60 min after administration suffered nociceptive stimulation, alcohol aversions were also detected in terms of alcohol ingestion patterns and ethanol odor aversions (Experiment 3). These results appear to indicate that orosensory ethanol processing takes place during acute ethanol intoxication probably due to ethanol elimination via respiration, salivation, and/or hematogenic olfaction processes.     

Odor-aversion learning in neonate rat pups: the role of duration of exposure to an odor

Five experiments employed 2- and 10-day-old rat pups to examine the ontogeny of odor-aversion learning. When duration of exposure to the to-be-conditioned odor stimulus (CS) was long, Experiments 1 and 2 demonstrated substantial and similar aversions across the two ages regardless of whether the toxin unconditioned stimulus (US) occurred simultaneously with, immediately before, immediately after, or some time after the CS. These experiments, therefore, did not support the claim of Rudy and Cheatle (Ontogeny of Learning and Memory. Hillsdale, N.J.: Erlbaum (1979) ) that the 2-day old pup suffers a deficit in its learning about the relation between successively presented events. The remaining experiments showed, however, that the 2-day-old pups did not acquire an aversion to a briefly presented CS, that the failure to condition this CS was not due to its overshadowing by contextual cues, and that the 10-day-old pup did acquire an aversion to that CS. The experiments have revealed, therefore, that there are age-related differences in the rat pup's ability to learn about the relation between a briefly-presented CS and the effects of a US.     

Simple disturbance of the dam in the neonatal period can alter haloperidol-induced catalepsy in the adult offspring

Three experiments were performed to determine whether apparently minimal disturbances of dams and litters would influence haloperidol-induced akinesia. In Experiment I, Long-Evans hooded rats (a) were left unmanipulated, (b) received nestcage relocation and observation, (c) received nestcage relocation/observation and maternal separation, or (d) received nestcage relocation/observation and pup handling. The male adult offspring received open-field testing and later received forepaw-on-dowel catalepsy testing following saline, morphine (20 mg/kg), or haloperidol (2 mg/kg). In Experiment II, hooded rats received (a) no manipulation, (b) nestcage relocation, (c) maternal separation, or (d) pup handling. At weaning, dams were tested in the open-field. Activity wheel locomotion of the offspring was assessed following saline or haloperidol for 3 days/week for 3 weeks; then, 5 and 7 days later, rats received haloperidol (0.5 mg/kg) and catalepsy testing. In both experiments, manipulations involving the dam reduced the offsprings' haloperidol-induced catalepsy, but, in Experiment II, a history of haloperidol administration distinguished between the effects of nestcage relocation and maternal separation. In Experiment III, Swiss albino mice received (a) no treatment, (b) nestcage relocation and maternal separation, (c) relocation/separation and mild cold stress of pups, (d) relocation/separation and pup handling, or (e) relocation/separation and severe cold stress of pups. Adult male mice received saline or haloperidol (2.5 mg/kg) and inclined grid catalepsy testing. Mice receiving relocation/separation and mice receiving relocation/separation and severe cold stress showed enhanced catalepsy versus control mice. Thus, it was concluded that seemingly innocuous events in infancy can influence the intensity of extrapyramidal side effects of neuroleptics in adulthood.     

The Influence of Retention Interval on the US Preexposure Effect: Changes in Contextual Blocking over Time

A number of studies manipulating the length of the interval between conditioning and testing indicate spontaneous recovery from overshadowing, suggesting that certain instances of overshadowing represent a deficit in memory retrieval rather than a failure of animals to form an association between the overshadowed stimulus and the US. The present series of experiments examined the influence of lengthening the retention interval on blocking, another stimulus selection phenomenon that is typically interpreted as an acquisition deficit. The results indicated that when subjects were tested shortly (3 days) after training conditioning to a taste blocked subsequent conditioning to an odor conditioned in compound with that taste (Experiment 1), whereas prior conditioning to an odor did not block subsequent conditioning to a taste conditioned in compound with that odor (Experiment 2). This pattern of results was essentially unchanged when testing occurred at a longer (21-day) retention interval. However, there was evidence of a US preexposure effect in Experiment 2 when subjects in the US ONLY control condition were tested at the 3-day retention interval, but not when testing occurred 21 days after conditioning. Experiments 3 and 4 examined whether this loss of the US preexposure effect over time might actually represent a change in the degree of contextual blocking as the retention interval is lengthened. Exposure to the conditioning context either during the interval between Phase 1 and Phase 2 of conditioning (Experiment 3) or prior to Phase 1 of conditioning (Experiment 4) alleviated this US preexposure effect suggesting that the loss of the US preexposure effect as the retention interval is lengthened observed in Experiment 2 is due to changes in the degree of blocking by contextual stimuli over time. The results are discussed in terms of differential susceptibility of forgetting of two functional roles played by a contextual stimuli in the current situation-context as a CS and context as a retrieval cue for other CS-US associations.     

Effects of preexposure flavor concentration on conditioned aversion and neophobia

In Experiment 1, 128 experimentally naive, water-deprived rats (Rattus norvegicus) received pretraining access to either 0.25 or 1.5% saccharin, distilled water, or 2.0% saline, followed either by a pairing of 0.25 or 1.5% saccharin with an intraperitoneal injection of 0.15 M lithium chloride (LiCl) or by a pairing of distilled water with LiCl. Preexposure to either saccharin concentration reliably reduced conditioned aversion effects to 0.25% saccharin, relative to that for preexposure to distilled water or saline. But only preexposure to 1.5% saccharin reduced aversion effects to that concentration. In Experiment 2, 48 naive, water-deprived rats received preexposure procedures as in Experiment 1. Afterwards, the rats were tested for neophobia to 0.25 or 1.5% saccharin. Neophobia was reliably greater to the 1.5% concentration. However, preexposure to either saccharin concentration obliterated evidence for neophobia to saccharin, relative to that following preexposure to distilled water or saline.     

Taste aversions to mother's milk: the age-related role of nursing in acquisition and expression of a learned association.

The development of taste aversion learning to novel cues contained in mother's milk was examined in rat pups. Pups receiving distinctive mild by experimenter-delivered oral infusions followed by toxicosis formed an aversion to the dam's diet. Robust aversions were learned as early as Day 10 and were retained for at least 11 days. When the same distinctive milk was obtained directly from a foster mother through nursing, however, only weanling-age pups (over 20 days) formed an aversion. X-ray analysis of nipple location in the mouths of suckling pups suggested that pups between the ages of 10 and 21 days receive milk at a similar tongue locus. Flavored milk was then delivered at specific time intervals in controlled quantities through tongue cannulas implanted at loci corresponding to the nipple position shown by the X-rays. Cannulated preweanling pups that were attached to a nipple during milk delivery failed to associate the taste cue with illness, whereas both preweanlings off the nipple and weanlings on the nipple acquired aversions to the taste cue in the milk. The evidence obtained in these experiments suggests that pups of all ages are incapable of expressing a taste aversion in a nursing situation and that preweanling pups in particular are also dificient in acquiring aversions within a suckling context. The inability of preweanling pups to acquire taste aversions in a nursing situation appears to result from a failure to associate taste cues with illness rather than a failure to detect taste cues obtained from a nipple.     

Representation-mediated extinction of conditioned flavor aversions

Conditioned flavor aversions were extinguished by presenting without consequence auditory stimuli that had been previously paired with the aversive flavor. In Experiment 1, rats that received tone-sucrose pairings, then sucrose-lithium chloride (LiCl) pairings, and finally repeated tone-alone presentations showed greater sucrose consumption in subsequent testing than rats that received similar sucrose-LiCl pairings and tone-alone presentations but no initial tone-sucrose pairings. Experiment 2 demonstrated the stimulus specificity of the mediated extinction observed in Experiment 1. In Experiment 3, rats that received first-order light-food and second-order tone-light pairings prior to sucrose-LiCl pairings did not show greater subsequent sucrose consumption when extinction of the second-order tone intervened. These results suggest that conditioned stimulus (CS)-evoked representations of events can substitute for those events themselves in the extinction of previously established associations.     

Acquisition and extended retention of a conditioned taste aversion in preweanling rats

The time course of memory decay for infant rats may shed light on the processes responsible for infantile amnesia. A taste aversion conditioning procedure appropriate for both neonatal and adult rats was employed in four experiments to investigate the ontogeny of extended retention. In Experiment 1, rats trained at 1, 10, 20, or 60 days of age were tested for retention of the taste aversion 25 days later. At testing, only those rats conditioned when 20 or 60 days old demonstrated significant taste aversions. Experiments 2 and 3 established that rats 14-15 days old and older were able to retain significant taste aversions following a 25-day retention interval. Younger rats did, however, acquire and retain the aversion for several days and showed a gradual retention loss over progressively longer retention intervals (Experiment 4). These findings suggest that preweanling rats demonstrate initial consolidation, storage, and retrieval of conditioned taste aversions. It is only after this initial period that retention deficits become evident.     

Postacquisition injection of tetrodotoxin into the parabrachial nuclei elicits partial disruption of passive avoidance reaction in rats.

The recent discovery that post-trial functional blockade of the parabrachial nuclei by intracerebral injection of 10 ng tetrodotoxin (TTX) disrupts acquisition of conditioned taste aversion (CTA) (Ivanova & Bures, 1990a,b) has prompted attempts to ascertain the role of this structure in other types of inhibitory learning. In Experiment 1, rats with implanted parabrachial cannulae were trained in a step-through avoidance task and received bilateral TTX (2 x 10 ng) immediately after the acquisition trial; they displayed significantly weakened avoidance of the shock compartment 2 days later. In Experiment 2, rats were anesthetized with pentobarbital (50 mg/kg) immediately after passive avoidance acquisition and received parabrachial TTX 15 min later; whereas anesthesia alone left the passive avoidance reaction (PAR) unaffected, TTX elicited similar disruption as in unanesthetized animals. In Experiment 3, TTX was injected in anesthetized animals 0, 1, 2, or 4 days after PAR acquisition. The amnesic effect was significant when the acquisition-TTX delay had been prolonged to 24 but not to 48 or 96 h. Since CTA is disrupted by reversible blockade of parabrachial nuclei and of the adjacent reticular formation elicited up to 4 days after acquisition (Ivanova & Bures, 1990b), PAR seems to be impaired to a lesser degree and for a shorter time than CTA by similar TTX treatment.     

Taste preconditioning augments odor-aversion learning

On the basis of previous work that has shown a taste can potentiate odor-aversion conditioning in AX+ conditioning, 6 experiments used rats to examine the effects of pairing a preconditioned taste (A) with a novel odor cue (X) in an A+/AX+ aversion conditioning design. Experiments 1A and 1B demonstrated that a preconditioned taste produced a robust odor aversion that was significantly stronger than a potentiated odor aversion. The results of Experiment 2 showed that the robust odor aversion produced by A+/AX+ conditioning was not the result of the potentiated odor aversion summating with generalization from the taste aversion. The augmented odor aversion was produced only when the taste and odor stimuli were presented simultaneously (Experiment 3) and the preconditioned taste aversion was intact at compound conditioning (Experiment 4). Pairing a novel odor with a preconditioned taste was not sufficient to condition an aversion to odor (Experiment 5), although other results implicated a role for an association between odor and taste in the odor augmentation effect (Experiment 6). The present results have implications for current models of taste + odor interactions in flavor-aversion conditioning.     

Second-order conditioning with diffuse auditory reinforcers in the pigeon

Pigeon subjects were used in five experiments investigating second-order conditioning with visual second-order and diffuse auditory first-order stimuli. Experiment 1 used a discriminative conditioning procedure to demonstrate reliable and substantial second-order conditioning with these stimuli. In Experiments 2 and 3, extinction of the auditory first-order stimulus after second-order conditioning had little effect upon responding to the second-order stimulus, when compared to a stimulus whose reinforcer was maintained. Experiment 4 compared directly the susceptibility of second-order responding to extinction of the first-order reinforcing stimulus as a function of the modality of that reinforcer. When a visual second-order stimulus was paired with a visual reinforcer, and the response to the latter was extinguished, then second-order responding was greatly reduced relative to control levels. In contrast, when that reinforcer was an auditory stimulus, second-order responding was not affected by the current value of the first-order stimulus after conditioning. Finally, in Experiment 5 the auditory stimulus was established as a reinforcer through discriminative-operant training. Following second-order conditioning, extinction of responding to this stimulus again had little impact upon responding to the second-order stimulus with which it had been paired. These results are discussed in terms of previous work on second-order conditioning with rat and pigeon subjects.     

Conditioned taste aversion is enhanced when the unconditioned stimulus is presented in a different context

Using a conditioned taste aversion procedure with rats as the subjects, two experiments examined the effect of presenting a conditioned stimulus (CS saccharin solution) in one context followed by an unconditioned stimulus (US LiCl) in a different context. Experiment 1 showed that animals which received the above-mentioned procedure (Group D) showed a more marked conditioned aversion to the CS than animals which were given both the CS and the US in the same context (Group S). Experiment 2 found that in both Group D and Group S, aversion to the CS increased when the subjects were exposed to the conditioned context after the conditioning. These findings supported the argument that the strength of the CS-US association acquired during conditioning is compared with that of the context-US to determine the magnitude of aversion revealed to the CS.