A temporal-specific and transient cAMP increase characterizes odorant classical conditioning

Increases in cyclic adenosine monophosphate (cAMP) are proposed to initiate learning in a wide variety of species. Here, we measure changes in cAMP in the olfactory bulb prior to, during, and following a classically conditioned odor preference trial in rat pups. Measurements were taken up to the point of maximal CREB phosphorylation in olfactory bulb mitral cells. Using both drug and natural unconditioned stimuli we found effective learning was associated with an increase in cAMP at the end of the conditioning trial, followed by a decrease 5 min later. This early timing of a transient cAMP increase occurred only when the odor was paired with an effective drug or natural unconditioned stimulus (US). The data support the hypothesis that the rate of adenylate cyclase activation is enhanced by pairing calcium and G-protein activation and that the timing of transient cAMP signaling is critical to the initiation of classical conditioning.     

CLASSICAL CONDITIONING OF COURTING BEHAVIOR IN THE JAPANESE QUAIL,COTURNIX COTURNIX JAPONICA1

Male Japanese quail were classically conditioned to display courting behavior at the sound of a buzzer, a previously neutral stimulus. The buzzer (conditioned stimulus) was paired with the presentation of a female quail (unconditioned stimulus) for a number of trials. The courting display, invariably elicited from the male by the presentation of the female, began to appear in part to the conditioned stimulus as early as the fifth pairing. All components of the display were elicited from all birds by the conditioned stimulus alone within 32 pairings of the conditioned and unconditioned stimuli.     

Pentylenetetrazole as an unconditioned stimulus for olfactory and contextual fear conditioning in rats

The association of five footshocks with a neutral odor is able to establish an olfactory fear conditioning in rats. The present study sought to investigate whether the systemic administration of pentylenetetrazole (PTZ; 3.75–15 mg/kg) would turn the coffee odor in a conditioned stimulus in the fear conditioning paradigm. The results showed that rats started to display risk assessment and avoidance after PTZ (15 mg/kg)–coffee odor pairing. When three mild footshocks (0.4 mA for 2 s) were delivered during this pairing, the conditioned response exhibited was greater than before. In both cases, however, pretreatment with the benzodiazepine midazolam (MDZ. 0.5 mg/kg i.p.) fully counteracted the expression of these defensive behaviors. Moreover, after being paired with 15 mg/kg of PTZ alone or combined with footshocks, the coffee odor was able to promote a new fear conditioning related to the context where it was re-exposed. The present findings point out the usefulness of PTZ as an unconditioned stimulus to promote fear conditioning to olfactory and contextual cues in rats.     

pCREB in the neonate rat olfactory bulb is selectively and transiently increased by odor preference-conditioned training

Early olfactory preference learning in rat pups occurs when novel odors are paired with tactile stimulation, for example stroking. cAMP-triggered phosphorylation of cAMP response element binding protein (pCREB) has been implicated as a mediator of learning and memory changes in various animals (Frank and Greenberg 1994). In the present study we investigate whether CREB is phosphorylated in response to conditioned olfactory training as might be predicted given the proposed role of the phosphorylated protein in learning. On postnatal day 6, pups were trained for 10 min using a standard conditioned olfactory learning paradigm in which a conditioned stimulus, Odor, was either used alone or paired with an unconditioned stimulus, Stroking (using a fine brush to stroke the pup). In some instances stroking only was used. The pups were sacrificed at 0, 10, 30, or 60 min after the training. Using Western blot analysis, we observed that the majority of olfactory bulbs in conditioned pups (Odor + Stroking) had a greater increase in pCREB activation at 10 min after training than pups given nonlearning training (Odor only or Stroking only). The phosphorylated protein levels were low at 0 min and at 60 min after training. This is in keeping with the slightly delayed and short-lived activation period for this protein. The localization of pCREB increases within the olfactory bulb as seen by immunocytochemistry. Naive pups were not exposed to odor or training. There was a significantly higher level of label in mitral cell nuclei within the dorsolateral quadrant of the bulb of pups undergoing odor-stroke pairing. No significant differences were observed among nonlearning groups (Naive, Odor only, or Stroking only) or among any training groups in the granule or periglomerular cells of the dorsolateral region. The localized changes in the nuclear protein are consistent with studies showing localized changes in the bulb in response to a learned familiar odor. The present study demonstrates that selective increases in pCREB occur as an early step following pairing procedures that normally lead to the development of long-term olfactory memories in rat pups. These results support the hypothesized link between pCREB and memory formation.     

Reflex conditioning in a spinal man.

A man with a completely transected spinal cord and a spastic neurogenic bladder was conditioned to void upon the presentation of an external stimulus. A classical conditioning paradigm was employed in which strong abdominal shock (unconditioned stimulus) was paired with an initially neutral mild electrical stimulation of the thigh (conditioned stimulus; CS). After the pairing trials, a reliable conditioned response of urination was elicited by the CS alone. The conditioned response did not extinguish over time, and the procedure left the bladder with clinically safe residual amounts of urine. Practical implications of the conditioning technique for the treatment of spastic neurogenic bladder conditions in spinally injured patients are discussed, as is the theoretical significance of conditioning at the reflex level in the absence of cortical involvement.     

Further evidence against the classical conditioning model of McCollough effects

McCollough effects (MEs) have been portrayed by Murch (1976) as classically conditioned responses which result from the pairing of color (unconditioned stimulus) with line orientation (conditioned stimulus). In the present experiment, we sought evidence that ME characteristics resembled those of undisputed conditioned responses. According to Rescorla (1968) and others, conditioned responses may not develop if the unconditioned stimulus is presented in the absence as well as in the presence of the conditioned stimulus. However, in our experiment, MEs induced by inspection sequences containing presentations of color alone in addition to color/ contour pairs were not weaker than controls. This discrepancy challenges the applicability of the conditioning model to MEs.     

Sign- versus goal-tracking: effects of conditioned-stimulus-to-unconditioned-stimulus distance.

Three pigeons were exposed sequentially across experimental phases to five different distances between the conditioned stimulus and the site of the unconditioned stimulus in a sign-/goal-tracking procedure. A computer-controlled tracking system provided a continuous record of the bird's position by continuously monitoring the location of the bird's head in three-dimensional space. It was found that birds sign-tracked (i.e., approached the conditioned stimulus) when the conditioned stimulus was closest to the site of the unconditioned stimulus, goal-tracked (i.e., approached the site of the unconditioned stimulus in the presence of the conditioned stimulus) when the conditioned stimulus was farthest from the site of the unconditioned stimulus, and engaged in both sign- and goal-tracking (or something intermediate) at intermediate conditioned-stimulus-to-unconditioned-stimulus distances. When both sign- and goal-tracking occurred, the former tended to occur in the first half and the latter in the second half of the interval in which the conditioned stimulus was present. The results suggest (a) whether sign- or goal-tracking (or both) occurs is a function of the distance of the conditioned stimulus from the site of the unconditioned stimulus, (b) the fact that pigeons but not rats have been found to sign-track consistently throughout the duration of the conditioned stimulus may be due to quantitatively rather than qualitatively different effects of conditioned-stimulus-to-unconditioned-stimulus distance across species (i.e., a "short" conditioned-stimulus-to-unconditioned-stimulus distance for a pigeon may be a "long" one for a rat), and (c) sign- and goal-tracking may be competing behavioral tendencies that can (e.g., at intermediate conditioned-stimulus-to-unconditioned-stimulus distances) cancel each other out. The findings lend support to theories that specify an interaction between phylogenetic and reinforcement variables in determining whether sign- or goal-tracking will occur in any given experimental preparation.     

Retention of a classically conditioned reflex response in spinal cat

Retention of classically conditioned flexion reflex facilitation was examined in unanesthetized, decerebrate, acute spinal cats. Flexion reflex facilitation, recorded from the tibialis anterior muscle, was obtained by pairing saphenous nerve stimulation (the conditioned stimulus) with superficial peroneal nerve stimulation (the unconditioned stimulus). The flexion reflex declined in control animals receiving the same number of nerve stimuli over the same time span, but in an explicitly unpaired sequence. To investigate retention, conditioned stimuli were presented at 5-min intervals following acquisition for a 2 1/2-h period. During this time a significant difference between conditioning and control groups was maintained even to the last trial, with no indication that the difference was subsiding over time. The results support the possibility that a classical conditioning paradigm applied to the spinal cord can induce alterations in spinal reflexes of long duration. Furthermore, the results appear to rule out post-tetanic potentiation as a mechanism producing the observed long-term effects.     

Changing likes and dislikes through the back door: The US-revaluation effect

US-revaluation refers to the observation that subsequent changes in the valence of an unconditioned stimulus (US) after pairing it with a neutral, conditioned stimulus (CS) also changes the valence of the associated CS. Experiment 1 found evidence for the US-revaluation effect using an unobtrusive measure of evaluation. However, US-revaluation effects were more pronounced for positive-to-negative compared to negative-to-positive revaluations. Experiment 2 replicated this finding for self-reported evaluations, further showing that US-revaluation effects are stable over time and independent of explicit memory for the revaluating information. Using a modified paradigm, Experiment 3 ruled out method-related explanations for these findings and showed that changes in CS evaluations are correlated with parallel changes in US evaluations. These findings encourage the view of evaluative conditioning as an instance of stimulus–stimulus (S–S) rather than stimulus-response (S–R) learning. Implications for basic and applied research are discussed.     

Inhibition and superconditioning

Superconditioning is said to occur when learning an association between a conditioned stimulus (CS) and unconditioned stimulus (US) is facilitated by pairing the CS with the US in the presence of a previously established conditioned inhibitor. Previous demonstrations of superconditioning have been criticized because their control conditions have allowed alternative interpretations. Using a within–subjects autoshaping procedure, the present study unambiguously demonstrated superconditioning. The results support the view that superconditioning is the symmetric opposite of blocking.     

味觉厌恶性条件反射与条件反射性免疫抑制的研究

以环磷酰胺的注射为非条件刺激（ＵＣＳ）,糖精水的摄入为条件刺激（ＣＳ）,并以血白细胞和淋巴细胞数量及脾淋巴细胞的转化反应为免疫指标,糖精水的饮用量为行为指标,通过改变条件刺激与非条件刺激结合的次数来观察条件反射性厌恶行为与条件反射性免疫抑制的反应。结果表明不管是一次性的还是两次性的ＣＳ－ＵＣＳ结合训练都能使动物明显地建立起味觉厌恶性条件反射,然而条件反射性免疫抑制只在两次性ＣＳ－ＵＣＳ结合训练后才     

Olfactory aversive conditioning in the newborn (3-hr-old) rat impairs later suckling for water and milk

An olfactory conditioning paradigm tested whether newborn rats can acquire a conditioned aversion to olfactory events associated with their first postnatal meal 3-5 hr after birth. Exposure to lemon odor (conditioned stimulus [CS]) paired with intraoral infusions of 0.1% quinine (unconditioned stimulus) resulted in explicit conditioning. Responsiveness to a surrogate nipple providing water in the presence of the CS was significantly lower than the 3 control conditions. The conditioning dramatically suppressed responsiveness to a surrogate nipple providing milk, which normally is expressed voraciously in terms of sustained nipple attachment and milk intake. These findings suggest that as early as 3-5 hr after birth newborn rats are capable of aversive conditioning to odors in the context of suckling behavior.     

Discriminative conditioning alters food preferences in the leech, Haemopis marmorata

The feeding behavior of the carnivorous leech, Haemopis marmorata, was aversively trained in a discriminative classical conditioning task. Two conditioned stimuli were used: One consisted of a food (chicken or liver) paired with an unconditioned stimulus of quinidine (bitter chemical); the other consisted of the alternate food presented in an unpaired relationship with the quinidine. Training consisted of alternating exposures to the two conditioned stimuli. Testing consisted of the simultaneous presentation of a conditioned stimulus food and a neutral food, beef. The percentages of responding to the conditioned stimuli were tabulated. Haemopis could discriminate between the conditioned stimuli. As a result of pairing a food with quinidine, the leeches selectively reduced their preference for that paired food, while they did not alter their preference for the unpaired food.     

Classical conditioning of eyelid and mystacial vibrissae responses in conscious mice

The murine vibrissae sensorimotor system has been scrutinized as a target of motor learning through trace classical conditioning. Conditioned eyelid responses were acquired by using weak electrical whisker-pad stimulation as conditioned stimulus (CS) and strong electrical periorbital stimulation as unconditioned stimulus (US). In addition, conditioned vibrissal protraction was obtained pairing either weak electrical whisker-pad stimulation or a tone as CS, with a strong electric shock delivered in the whisker-pad as US. This finding suggests that evolutionary pressure has selected a sensorimotor system capable of constructing conditioned responses on the basis of temporal relationships of stimuli, independently of any putative functional purpose.     

条件反射性免疫抑制激活过程中下丘脑核团c-fos的表达

利用经典条件反射性免疫抑制的动物模型,以糖精水为条件刺激（CS）,免疫抑制剂-环磷酰胺为非条件刺激 （UCS）,观测两次CS-UCS结合训练后,再次条件刺激诱发条件反射性免疫抑制作用的动态改变,获得条件反射性免疫抑制和味觉厌恶性条件反射各自保持的情况,并在此基础上,采用c-fos免疫组化技术, 进一步观察再次条件刺激诱发条件反射性免疫抑制反应时大鼠下丘脑各核团内FOS蛋白的表达情况。结果表明,条件反射性免疫抑制作用在训练后第5天较强,第30天基本消失,而味觉厌恶性条件反射始终稳定保持到第30天。进一步研究显示出,下丘脑室旁核FOS蛋白表达在第5天非常密集,而第30天几乎没有表达,与细胞免疫功能改变在时程和趋势上具有一致性。通过FOS蛋白表达时程差异比较,提示下丘脑室旁核可能是CNS内介导CS诱导的免疫抑制效应的重要核团。     

Using pavlovian higher-order conditioning paradigms to investigate the neural substrates of emotional learning and memory

In first-order Pavlovian conditioning, learning is acquired by pairing a conditioned stimulus (CS) with an intrinsically motivating unconditioned stimulus (US; e.g., food or shock). In higher-order Pavlovian conditioning (sensory preconditioning and second-order conditioning), the CS is paired with a stimulus that has motivational value that is acquired rather than intrinsic. This review describes some of the ways higher-order conditioning paradigms can be used to elucidate substrates of learning and memory, primarily focusing on fear conditioning. First-order conditioning, second-order conditioning, and sensory preconditioning allow for the controlled demonstration of three distinct forms of memory, the neural substrates of which can thus be analyzed. Higher-order conditioning phenomena allow one to distinguish more precisely between processes involved in transmission of sensory or motor information and processes involved in the plasticity underlying learning. Finally, higher-order conditioning paradigms may also allow one to distinguish between processes involved in behavioral expression of memory retrieval versus processes involved in memory retrieval itself.     

Classical conditioning of hippocampal theta patterns in the rat.

Electrical stimulation (88 HZ) of the lateral hypothalamus elicited a sustained theta response at hippocampal recording sites of rats immobilized with succinylcholine. By pairing this unconditioned stimulus with a 10-sec presentation of a light, conditioned theta responses were demonstrated in as few as 40 trials. Spectral analysis of hippocampal bioelectric patterns during acquisition, extinction, and reconditioning indicated that the earliest change as a result of conditioning is a loss of power in EEG frequency below 8 HZ, followed by the development of a peak at 8 HZ with further conditioning. Extinction was associated with an increase in power in the frequencies below 8 HZ. When the conditioned rats were tested in the absence of the neuromuscular blocking agent, the conditioned stimulus elicited a theta response that was associated with slow motor activity on 70% of the trials.     

Factors influencing responding under multiple schedules of conditioned and unconditioned reinforcement

Two experiments examined pigeons' responses under multiple schedules of conditioned and unconditioned reinforcement. In one component, responses produced food according to a fixed-interval schedule; in a second component, responses produced brief stimuli according to a fixed-ratio schedule. When brief-stimulus presentations were paired with food in the first component, rates in the second component were usually higher than 10 responses per minute. When pairing in the first component was eliminated, responding continued to be maintained in the second component. Elimination of food presentation from the first component substantially decreased responding in the second component, even though the brief stimulus had not been paired with food. Experiment II demonstrated that response rate was affected by the duration of both the second component and the brief stimulus. The results suggest that three conditions are important in maintaining responding with brief-stimulus presentations: (1) pairing the brief stimulus, at least initially, with food, (2) maintaining unconditioned reinforcement in one component, and (3) employing optimal brief-stimulus and component durations.     

Neuronal unit activity in the abducens nucleus during classical conditioning of the nictitating membrane response in the rabbit (Oryctolagus cuniculus).

Neuronal unit activity was recorded from the abducens (6th nerve) nucleus, the "final common path," during classical conditioning of the nictitating membrane (NM) response in the rabbit, with the use of a tone conditioned stimulus, an air puff unconditioned stimulus (UCS), 250-msec interstimulus interval, and 60-sec intertrial interval. Animals were given 2 days of conditioning training (104 trials in eight blocks per day) and 1 day of extinction. Control animals were given comparable periods of stimulus presentations, explicitly unpaired. Activity of small clusters of units--"multiple unit" recording--was compared with the amplitude-time course of the NM response. Between-blocks comparisons of neural and behavioral responses indicated an essentially perfect correlation during acquisition of the conditioned response (Day 1, r = .99; Day 2, r = .98) and a slightly lower correlation during extinction (r = .93) for the conditioning animals. Within-blocks comparisons indicated a close correspondence between the histograms of unit activity and the amplitude-time course of the NM response for the conditioning animals in all phase of training and for the control animals in the UCS trial blocks.     

An animal model of fetishism

An animal model of sexual fetishism was developed with male Japanese quail based on persistence of conditioned sexual responding during extinction to an inanimate object made of terrycloth (Experiments 1 and 3). This persistent responding occurred only in subjects that came to copulate with the terrycloth object, suggesting that the copulatory behavior served to maintain the fetishistic behavior. Sexual conditioning was carried out by pairing a conditioned stimulus (CS) with the opportunity to copulate with a female (the unconditioned stimulus or US). Copulation with the CS object and persistent responding did not develop if the CS was a light (Experiment 1) or if conditioning was carried out with a food US (Experiment 2). In addition, subjects that showed persistence in responding to the terrycloth CS did not persist in their responding to a light CS (Experiment 3). The results are consistent with the hypothesis that conditioned copulatory behavior creates a form of self-maintenance that leads to persistent responding to an inanimate object. The development of an animal model of such fetishistic behavior should facilitate experimental analysis of the phenomenon.