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Conditioned alterations of specific blood leukocyte subsets are reconditionable.

Behavioral conditioning has the ability to produce changes in immune function. However, it is unknown whether conditioned changes of immune function can be recalled on multiple occasions. To address this issue we paired a novel saccharin drinking solution with intraperitoneal cyclosporin A (CsA) injection in rats. Saccharin re-presentation produced a reduction in splenocyte proliferation that mirrored the effect of CsA. Such functional changes were paralleled by a significant conditioned leukopenia in peripheral blood, which opposed the leukocytosis induced by CsA. Using the conditioned leukopenia in blood as a 'diagnostic window' of conditioned immunosuppression, the maintenance of CsA-induced changes was investigated by examining blood samples collected repeatedly. Experiments on the same group of animals over a period of 1 year showed that the conditioned leukopenia was reproducible on multiple occasions by reimplementing either the whole conditioning paradigm or reexposure to the saccharin solution only. These results demonstrate that behaviorally conditioned alterations of immune parameters are maintained in subsequent trials, indicating the potential clinical feasibility of behavioral conditioning procedures.

Animals↗

Serotonergic depletion increases conditioned suppression to background stimuli in the rat.

Dark Agouti rats were lesioned by intra-ventricular injection of 5,7-dihydroxytryptamine (DHT) and, 2 weeks later, learning was tested in a conditioned suppression of drinking procedure. Lesioned and vehicle-injected control rats were conditioned with a discrete stimulus (tone or light conditioned stimulus, CS) twice paired with footshock (unconditioned stimulus), with or without a 30-s trace interval between these events to produce strong and weak learning conditions (a trace conditioning effect). During this conditioning session, the alternate stimulus (light or tone) was presented continuously in the background. Since the 5,7-DHT lesion also reduced the baseline licking response in the experimental chambers, we used drinking during the first minute, when this non-specific effect was minimal, as the dependent variable. We tested conditioning to target CS and to the alternative experimental background stimulus in exactly the same way in the same rats. We found that a level of serotonergic depletion without any intrinsic action on the trace conditioning effect nevertheless increased conditioning to the alternative background stimulus, irrespective of trace interval or stimulus modality. Thus, for both light and tone stimuli, the effect of serotonergic depletion depended only on the discrete target versus diffuse background role of the stimulus in use. These findings have implications for the modification of human cognition by serotonergic drugs.

5,7-Dihydroxytryptamine↗

Methylphenidate can reduce selectivity in associative learning in an aversive trace conditioning task.

There are good grounds to expect that methylphenidate (MP) should enhance cognitive function. However, experimental evidence on this point is scant. The present study therefore examined the effects of MP on learning the association between a conditioned stimulus (CS, in this case, noise) and an unconditioned stimulus (UCS, in this case, footshock) in an aversive variant of a trace conditioning procedure. Learning was measured off-the-baseline as conditioned suppression of drinking (both latencies to drink, expressed as suppression ratios, and the amount drunk, expressed as the number of licks, in the presence of the CS). In addition to the measures of discrete cue conditioning, MP effects on contextual conditioning were measured as suppression to apparatus cues and an experimental background stimulus. MP was administered at 1 or 5 mg/kg prior to conditioning sessions. As attention deficit hyperactivity disorder (ADHD) has been characterized as involving a ;wide attentional window' (e.g. Shalev and Tsal, 2003), it was predicted that MP, as the treatment of choice for ADHD, should increase selectivity (narrowing the attentional window). This outcome would show as reduced levels of conditioning (compared to control rats) to less informative trace and contextual cues present during conditioning. Contrary to prediction, both 1 and 5 mg/kg MP increased learning about all the available stimuli, including the less informative trace CS and the background stimulus. These findings are consistent with reduced rather than increased selectivity in learning (because of increased rather than decreased conditioning to weak cues) under MP.

Animals↗

Responses of the circadian system of rats to conditioned and unconditioned stimuli.

The circadian systems of rodents respond to light pulses presented during the subjective night with phase shifts and altered cellular activity in the suprachiasmatic nuclei (SCN), including expression of immediate-early genes (IEGs) such as c-fos. A recent study showed that a nonphotic stimulus (an air disturbance generated by a fan) that does not normally induce the expression of c-fos-like immunoreactivity in the SCN of rats can be made to do so after being paired repeatedly with a light pulse in a Pavlovian conditioning paradigm. Furthermore, after conditioning (but not after noncontingent exposure to these stimuli), the fan also induced phase shifts in activity and body temperature rhythms comparable to those produced by light. The authors performed three experiments designed to replicate and extend these findings in rats. In experiment 1, rats were tested for conditioning effects of repeated pairings of a light pulse with a neutral air disturbance under a full photoperiod. In experiment 2, a modified conditioning paradigm was used in which a skeleton photoperiod served as both the entraining zeitgeber and the unconditioned stimulus. Animals in the paired and unpaired training conditions were exposed to both the light pulse and the air disturbance, but the air disturbance signaled the onset of light in the paired condition only. Phase shifts of wheel-running activity rhythms and gene expression in the SCN, intergeniculate leaflet, and paraventricular nucleus of the thalamus were assessed in animals following either of the training conditions or the control procedures. Experiment 3 assessed whether the air disturbance could entrain the circadian activity rhythms of rats with or without previous pairing with light in a classical conditioning paradigm. No evidence for classical conditioning, nor for unconditioned effects of the air disturbance on the circadian system, was found in these studies.

Air↗

Selective neurotoxic lesions of basolateral and central nuclei of the amygdala produce differential effects on fear conditioning.

In the fear conditioning literature, it is generally hypothesized that neurons in the basolateral amygdalar complex (BLA) (lateral and basal nuclei) support the formation of conditioned fear memory and project to neurons in the central nucleus (CeA) for the expression of conditioned fear responses. According to this serial processing-transmission view, damage to either BLA or CeA would comparably disrupt the expression of a variety of conditioned fear responses. In the present study, we further investigated the roles of BLA and CeA in fear conditioning by concurrently assessing freezing and 22 kHz ultrasonic vocalization (USV) as dependent measures of fear in rats. Selective neurotoxins, NMDA for the BLA and ibotenic acid for the CeA, were used to destroy intrinsic neurons [evidenced by thionin dye and NeuN (neuronal nuclei) antibody stainings] without damaging the fibers of passage (confirmed by myelin staining). During the 10 tone-footshock paired training, postshock freezing and USV responses were significantly impaired in BLA-lesioned animals, whereas CeA-lesioned animals exhibited only mild deficits. Similarly, conditioned fear responses assessed 24 hr after training were severely reduced in BLA-lesioned animals but not in CeA-lesioned animals. In contrast to ibotenic lesions of the CeA, small electrolytic lesions of the CeA strongly affected both postshock and conditioned freezing and USV. Together, these results do not support the currently espoused BLA-to-CeA serial processing-transmission view of fear conditioning. Instead, the expression of conditioned fear appears to primarily involve BLA projections that course through the CeA en route to downstream fear response structures.

Amygdala↗

Lesions of the basal amygdala block expression of conditioned fear but not extinction.

Although the role of the amygdala in acquisition of conditioned fear is well established, there is debate concerning the intra-amygdala circuits involved. The lateral nucleus of the amygdala (LA) is thought to be an essential site of plasticity in fear conditioning. The LA has both direct and indirect [via the basal nuclei; basal amygdala (BA)] projections to the central nucleus (Ce) of the amygdala, an essential output for fear behaviors. Lesions of the LA or Ce prevent acquisition of conditioned freezing to a conditioned stimulus, but BA lesions do not, suggesting that the BA is not normally involved in fear conditioning. If true, posttraining BA lesions should also have no effect. Replicating previous studies, we found that rats given electrolytic BA lesions before training acquired conditioned fear normally. They also showed normal long-term retention and extinction of conditioned fear. Unexpectedly, BA lesions made after training completely blocked expression of conditioned fear. Despite this deficit, lesioned rats were able to learn a new tone-shock association. Thus, although the LA-Ce system is sufficient for fear acquisition in the absence of the BA, it is not sufficient when the BA is present, suggesting that the BA is an important site of plasticity in fear conditioning. The pattern of lesion deficits we observed (after but not before training) might be explained by homeostatic mechanisms that balance plasticity over multiple inputs, regulating the influence of the BA and LA onto Ce output neurons.

Amygdala↗

Withdrawal from chronic nicotine administration impairs contextual fear conditioning in C57BL/6 mice.

The effects of acute nicotine administration (0.09 mg/kg nicotine), chronic nicotine administration (6.3 mg/kg/d nicotine for 14 d), and withdrawal from chronic nicotine administration on fear conditioning in C57BL/6 mice were examined. Mice were trained using two coterminating conditioned stimulus (30 s; 85 dB white noise)--unconditioned stimulus (2 s; 0.57 mA foot shock) pairings and tested 24 h later for contextual and cued fear conditioning. Acute nicotine administration enhanced contextual fear conditioning, chronic nicotine administration had no effect on contextual fear conditioning, and withdrawal from chronic nicotine administration impaired contextual fear conditioning. Plasma nicotine concentrations were similar after acute and chronic treatment and were within the range reported for smokers. During withdrawal, concentrations of nicotine were undetectable. An acute dose of nicotine (0.09 mg/kg) during withdrawal from chronic nicotine treatment reversed withdrawal-associated deficits in contextual fear conditioning. The results suggest that tolerance to the effects of nicotine on contextual fear conditioning develops with chronic nicotine treatment at a physiologically relevant dose, and withdrawal from this chronic nicotine treatment is associated with impairments in contextual fear conditioning. These findings provide a model of how the effects of nicotine on learning may contribute to the development and maintenance of nicotine addiction.

Animals↗

Effects of suspension of air-conditioning on airtight-type racks.

Although isolation racks are superior to open-type racks in terms of securing breeding conditions for laboratory animals, the contingency-proofing capability of the former has yet to be determined. Therefore, from the view of risk management, we studied the environmental change in isolation racks by forcibly suspending ventilation and air-conditioning and confirming the maximal time length for complete recovery to the original condition after restarting their operations. The isolation racks were placed in a room that was equipped with an independent air-conditioning system. When the inside condition of the racks reached 22-24 degrees C and 59-64% of relative humidity, the air-conditioning and ventilation were forcibly suspended and the subsequent temperature, relative humidity, ammonium and CO2 concentrations in the racks were measured over time. We found that after suspending the air-conditioning and ventilation, it took 40-60 min for temperature, and about 10 min for relative humidity to exceed the maximum values (temperature and relative humidity) referred to in the Showa 58 Nenban Guideline Jikken Doubutsu Shisetsu no Kenchiku oyobi Setsubi (Guidelines of buildings and facilities for experimental animals in Japan; Year 1983 edition). After 17 hr 25 min of the suspension of air-conditioning and ventilation, two rats were found dead. Then, the air-conditioning and ventilation were restarted. It took about 2 hr for temperature, and 50 min for relative humidity to regain the guideline values. The ammonium concentration stayed within the guideline value with a maximum concentration of 2 ppm in the experimental period, whereas the CO2 concentration was found to exceed 9% at the time of animal death.

Air Conditioning↗

Ideational conditioning: bridging the cognitive-behaviourist divide.

Classical conditioning has traditionally been referred to as a 'dumb form of learning'; however, psychological activity can be redefined in terms of the conditioned reflex by considering other more complex forms of conditioning, i.e., configural or semantic conditioning. In this paper it is argued that conditioning may underpin both human perception and consciousness and previous experiments involving perceptual conditioning are outlined. Recent experiments which involve a new paradigm of ideational conditioning have shown that cognitions can play a part in conditioning processes. It is shown here that ideational conditioning demonstrates unequivocally that thoughts alone are adequate stimuli for acquisition of behavioural responses and that these experiments may help to bridge the cognitive-behaviourist divide.

Arousal↗

Operant conditioning of the pain experience.

Two experiments were carried out to study operant conditioning of pain report. Further, it was investigated whether pain-related psychophysiological and psychological measures (skin conductance response and magnitude matching) could also be conditioned operantly. In both experiments subjects received 12 painful electric shocks of equal intensity. In Exp. 1 healthy subjects were assigned to either a control group or an up-conditioning group. Up-conditioning occurred by verbally rewarding increases in pain report and punishing decreases. Analyses indicated that up-conditioning of both pain report and the pain-related psychophysiological measures succeeded. To rule out alternative explanations of the results (attention shift towards pain and conditioning of anxiety) the verbal punishments were adjusted in Exp. 2. A down-conditioning group was also added. The attempt to replicate the results of Exp. 1 failed and down-conditioning of the pain report could not be established. These inconsistent results are most probably due to modified punishment of responses. The consequences for the results of Exp. 1 are discussed. Based on the results of post hoc analyses, some suggestions are made for operant conditioning studies of pain.

Adolescent↗

Behaviorally conditioned modulation of natural killer cell activity: enhancement of baseline and activated natural killer cell activity.

Natural killer (NK) cells comprise a sub-population of lymphocytes functionally defined by their ability to spontaneously lyse select tumor and virally infected cells. NK cell lytic function is sensitive to modulation by cytokines and neuroendocrine mediators. Behavioral conditioning trains an animal to cognitively associate a novel environmental cue (i.e. odor) with a physiologically active agent (i.e. Polylnosinic:PolyCytidilic acid, Poly I:C). Poly I:C induces interferon production resulting in activation of NK cells and enhanced NK cell lytic activity. Subsequent to behavioral conditioning, independent presentation of the odor should elicit similar responses (enhanced NK cell activity). We have shown that animals pre-exposed to the conditioning apparatus or manual restraint prior to behavioral conditioning demonstrate enhanced baseline NK cell activity or no effects respectively. To assess the influence of NK cell activation in conjunction with behavioral conditioning, we have re-presented the odor to animals with activated or baseline NK activity. Lymphocytes were then incubated for five days with IL-2 and cellular cytotoxic activity re-assessed. Behavioral conditioning significantly enhanced baseline and activated NK cell activity in animals previously exposed to the conditioning apparatus. No differences in lytic activity versus NK sensitive targets were observed following IL-2 activation. In contrast, animals manually restrained prior to conditioning showed no differences in baseline or in vivo activated NK activity, but previously non-activated lymphocytes stimulated with IL-2 demonstrated significantly higher lytic activity in conditioned animals re-presented with the odor. These results demonstrate central nervous system (CNS) modulation of NK cell activity and the presence of an interplay between cytokine activation and responsiveness to CNS immunoregulatory signals.

Animals↗

Effect of changing the frequency of conditioning tetanus on depressor responses evoked by stimulation of the aortic nerve.

The effect of changes in frequency of the conditioning tetanus on the magnitude of the testing depressor response was studied in rabbits anaesthetized with urethane. Conditioning and testing stimulations were applied to the same aortic nerve. The duration of the conditioning tetani was set at 3 and 60 sec and the interval between stimulations amounted to 40 and 120 sec. At the testing interval of 40 sec the increase in frequency both of short and long conditioning tetani reduces the magnitude of the testing response which attains a minimum at frequency of about 30 cycles/sec. Conditioning stimulations of higher frequency are gradually less effective and cause the testing response to increase. Similar depression is observed at the testing interval of 120 sec but only following long-lasting conditioning tetanus. Short conditioning trains at the testing interval of 120 sec facilitate the testing response. The frequency of the conditioning stimulation which produces the greatest reduction of the depressor response indicates the range of control exerted by the conditioning tetanus over the testing blood pressure effect. The size of this control is determined by the lowest level of depression and the highest value of facilitation of the testing response.

Animals↗

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.

Animals↗

Competition between ethanol-induced reward and aversion in place conditioning.

Previous place conditioning studies in mice have shown that injection of ethanol immediately before a conditioned stimulus (CS+) produces conditioned preference, whereas injection of ethanol immediately after CS+ produces conditioned aversion. In the present experiments, we examined the learning that occurs when ethanol is injected in "ambiguous" procedures that provide the opportunity for both types of conditioning. When ethanol was given midway through the CS (Experiments 1 and 2) or both before and after the CS (Experiment 3), the direction of place conditioning was the same as when mice were exposed only to whichever contingency occurred first (a primacy effect). That is, injection of ethanol in the middle of the CS conditioned aversion, whereas injection both before and after the CS conditioned preference. Because these results support the idea that ethanol elicits both aversive and rewarding effects, they are most consistent with conditioning theories that conceptualize unconditioned stimuli (USs) as events that can activate multiple representational components.

Animals↗

Lesions of the dorsal hippocampal formation interfere with background but not foreground contextual fear conditioning.

The effects of hippocampal lesions on the conditioning of fear responses (freezing responses) to contextual stimuli (static, continuously present stimuli) were examined in three conditioning paradigms: forward pairing of a phasic tone conditioned stimulus (CS) with a footshock unconditioned stimulus (US), unpaired presentations of the CS and US, or presentations of the US alone. All three procedures resulted in the acquisition of conditioned freezing to contextual stimuli. Lesions of the dorsal hippocampus prevented the acquisition of contextual conditioning in the Paired procedure, as reported previously, but not in the Unpaired or US Alone procedures. In the Paired procedure, static contextual cues occur in the background, with the phasic tone CS being the primary stimulus that enters into the association with the US. However, in the other two procedures, where there is no phasic CS, the primary associations with the US involve static contextual stimuli, which are therefore in the foreground. We refer to these types of contextual conditioning as background and foreground contextual conditioning, respectively, and argue that the hippocampus is only involved in background contextual conditioning. These results have implications for understanding both fear conditioning and hippocampal function.

Animals↗

The role of the primate amygdala in conditioned reinforcement.

Conditioned reinforcement refers to the capacity of a conditioned stimulus to support instrumental behavior by acquiring affective properties of the primary reinforcer with which it is associated. Conditioned reinforcers maintain behavior over protracted periods of time in the absence of, and potentially in conflict with, primary reinforcers and as such may play a fundamental role in complex social behavior. A relatively large body of evidence supports the view that the amygdala (and in particular the basolateral area) contributes to conditioned reinforcement by maintaining a representation of the affective value of conditioned stimuli. However, a recent study in primates (Malkova et al., 1997), using a second-order visual discrimination task, suggests that the amygdala is not critical for the conditioned reinforcement process. In the present study, excitotoxic lesions of the amygdala in a new world primate, the common marmoset, resulted in a progressive impairment in responding under a second-order schedule of food reinforcement. In addition, the responding of amygdala-lesioned animals was insensitive to the omission of the conditioned reinforcer, unlike that of control animals, for which responding was markedly reduced. In contrast, lesioned animals were unimpaired when responding on a progression of fixed-ratio schedules of primary reinforcement. These data confirm that the amygdala is critical for the conditioned reinforcement process in primates, and taken together with other recent work in monkeys, these results suggest that the contribution of the amygdala is to provide the affective value of specific reinforcers as accessed by associated conditioned stimuli.

Acoustic Stimulation↗

Standard delay eyeblink classical conditioning is independent of awareness.

P. F. Lovibond and D. R. Shanks (2002) suggested that all forms of classical conditioning depend on awareness of the stimulus contingencies. This article considers the available data for eyeblink classical conditioning, including data from 2 studies (R. E. Clark, J. R. Manns, & L. R. Squire, 2001; J. R. Manns, R. E. Clark, & L. R. Squire, 2001) that were completed too recently to have been considered in their review. In addition, in response to questions raised by P. F. Lovibond and D. R. Shanks, 2 new analyses of data are presented from studies published previously. The available data from humans and experimental animals provide strong evidence that delay eyeblink classical conditioning (but not trace eyeblink classical conditioning) can be acquired and retained independently of the forebrain and independently of awareness. This conclusion applies to standard conditioning paradigms; for example, to single-cue delay conditioning when a tone is used as the conditioned stimulus (CS) and to differential delay conditioning when the positive and negative conditioned stimuli (CS+ and CS-) are a tone and white noise.

Aged↗

Methamphetamine-induced conditioned place preference is facilitated by estradiol pretreatment in female mice.

Ovarian hormones were well documented to modulate the dopamine release in the central dopaminergic systems. The dopamine-releasing effects in the nucleus accumbens, a major target of the mesolimbicortical dopaminergic system, were closely associated with the reinforcing effects of two psychomotor stimulants, cocaine and methamphetamine. This study aimed to examine the sex differences in the cocaine- and methamphetamine-reinforcing behavior, conditioned place preference. In addition, the modulating effects of estradiol and progesterone on methamphetamine-induced conditioned place preference were investigated in both sexes of adult gonadectomized mice. There was no sex difference in the sensitivity to the cocaine (5 mg/kg)-induced conditioned place preference. However, female mice exhibited a more potent methamphetamine (1 mg/kg)-induced conditioned place preference than did male mice. Moreover, pretreatment with estradiol for two consecutive days before the beginning of the conditioning and throughout the four daily conditionings (0.47 microg/day for totally six days) effectively facilitated methamphetamine-induced conditioned place preference in gonadectomized female mice, but not in gonadectomized male mice. Progesterone, under a similar treatment regimen (0.47 microg/day for six consecutive days), did not alter the methamphetamine-induced conditioned place preference in either sex of gonadectomized mice. Taken together, we conclude that the facilitating effects of estradiol on methamphetamine-induced conditioned place preference could be sex-dependent with an eminent sensitivity associated with the adult female mice.

Animals↗