Variability in prescription opioid intake and reinforcement amongst 129 substrains.

Opioid abuse in the United States has reached epidemic proportions, with treatment admissions and deaths associated with prescription opioid abuse quadrupling over the past 10 years. Although genetics are theorized to contribute substantially to inter-individual variability in the development, severity and treatment outcomes of opioid abuse/addiction, little direct preclinical study has focused on the behavioral genetics of prescription opioid reinforcement and drug-taking. Herein, we employed different 129 substrains of mice currently available from The Jackson Laboratory (129S1/SvlmJ, 129X1/SvJ, 129S4/SvJaeJ and 129P3/J) as a model system of genetic variation and assayed mice for oral opioid intake and reinforcement, as well as behavioral and somatic signs of dependence. All substrains exhibited a dose-dependent increase in oral oxycodone and heroin preference and intake under limited-access procedures and all, but 129S1/SvlmJ mice, exhibited oxycodone reinforcement. Relative to the other substrains, 129P3/J mice exhibited higher heroin and oxycodone intake. While 129X1/SvJ exhibited the highest anxiety-like behavior during natural opioid withdrawal, somatic and behavior signs of precipitated withdrawal were most robust in 129P3/J mice. These results demonstrate the feasibility and relative sensitivity of our oral opioid self-administration procedures for detecting substrain differences in drug reinforcement/intake among 129 mice, of relevance to the identification of genetic variants contributing to high vs. low oxycodone reinforcement and intake.

Estrous odors and sexually conditioned neutral odors activate separate neural pathways in the male rat.

Olfactory stimuli play important roles in sexual behavior. Previous studies have demonstrated that both estrous odors and initially neutral odors paired with copulation influence the sexual behavior of male rats. The present study examines the pattern of neural activation as revealed by Fos immunoreactivity (Fos-IR) following exposure to bedding scented with either a neutral odor (almond) paired previously with copulation, estrous odors or no odor. Following exposure to estrous odors Fos-IR increased in the accessory olfactory bulb, medial amygdala, medial bed nucleus of the stria terminalis, medial preoptic area, ventromedial hypothalamus, ventral tegmental area, and both the nucleus accumbens core and shell. Conversely, following exposure to the sexually conditioned odor Fos-IR increased in the piriform cortex, basolateral amygdala, nucleus accumbens core, and the anterior portion of the lateral hypothalamic area. In addition, following exposure to almond odor Fos-IR increased in the main olfactory bulb independent of its pairing with copulation. These patterns of Fos-IR following exposure to estrous or sexually conditioned odors were not influenced by either the addition or omission of the other type of odor. These findings demonstrate that estrous and sexually conditioned odors are processed by distinct neural pathways and converge in the nucleus accumbens core, suggesting that this structure has a unique role in processing sexual stimuli of both pheromonal and olfactory natures.

Food-entrained circadian rhythms in rats are insensitive to deuterium oxide.

Rats anticipate a scheduled daily meal by entrainment of a circadian pacemaker separate from the light-entrainable circadian pacemaker located in the suprachiasmatic nuclei (SCN). The site and molecular mechanisms of the food-entrainable pacemaker are unknown. The intrinsic period (tau) of the SCN pacemaker is significantly lengthened by deuteriation. Sensitivity of food-entrained circadian rhythms to D(2)O (25% in drinking water) was evaluated in intact and SCN-ablated rats entrained to daily feeding schedules. In intact rats fed ad-libitum, D(2)O lengthened tau sufficiently to drive activity rhythms out of entrainment to the light-dark cycle. By contrast, food-entrained rhythms were surprisingly resistant to modulation by D(2)O. The mean daily onset time of food anticipatory activity in rats with complete SCN-ablations was not affected by up to 28 days of D(2)O intake. Transient delays and disruption of anticipatory activity were evident in intact and one partial SCN-ablated rat during D(2)O treatment, but these are interpretable as effects of coupling and/or masking interactions between a D(2)O-sensitive light-entrainable pacemaker, and a D(2)O-resistant food-entrained pacemaker. Differential sensitivity to D(2)O suggests diversity in the molecular mechanisms of food- and light-entrainable circadian pacemakers in mammals. D(2)O may have utility as a screening test to identify putative food-entrainable pacemakers from among those central and peripheral tissues that can express circadian oscillations of clock genes independent of the SCN.

Conditioning and sexual behavior: a review.

Sexual behavior is directed by a sophisticated interplay between steroid hormone actions in the brain that give rise to sexual arousability and experience with sexual reward that gives rise to expectations of competent sexual activity, sexual desire, arousal, and performance. Sexual experience allows animals to form instrumental associations between internal or external stimuli and behaviors that lead to different sexual rewards. Furthermore, Pavlovian associations between internal and external stimuli allow animals to predict sexual outcomes. These two types of learning build upon instinctual mechanisms to create distinctive, and seemingly "automated," patterns of sexual response. This article reviews the literature on conditioning and sexual behavior with a particular emphasis on incentive sequences of sexual behavior that move animals from distal to proximal with regard to sexual stimuli during appetitive phases of behavior and ultimately result in copulatory interaction and mating during consummatory phases of behavior. Accordingly, the role of learning in sexual excitement, in behaviors that bring about the opportunity to mate, in courtship and solicitation displays, in sexual arousal and copulatory behaviors, in sexual partner preferences, and the short- and long-term influence of copulatory experience on sexual and reproductive function is examined. Although hormone actions set the stage for sexual activity by generating the ability of animals to become sexually excited and aroused, it is each animal's unique experience with sexual behavior and sexual reward that molds the strength of responses made toward sexual incentives.

The development of olfactory conditioned ejaculatory preferences in the male rat. I. Nature of the unconditioned stimulus.

We have demonstrated previously that repeated pairing of a neutral odor with copulation produces a subsequent conditioned ejaculatory preference (CEP) for a female bearing that odor. Here we examine the copulatory components that comprise the unconditioned stimulus (UCS). In Experiment 1, male Long-Evans rats were allowed to copulate with scented females for nine sessions in which they achieved two ejaculations, one ejaculation plus the first intromission following the postejaculatory interval (PEI), one ejaculation without a PEI, or five intromissions without ejaculation. Only the males that achieved two ejaculations or one ejaculation plus the PEI displayed significant CEP. In Experiment 2, males were allowed to remain in the presence of the scented female without access to her after different amounts of copulatory stimulation. Under these conditions, both one and two ejaculations, but not five intromissions, supported the development of CEPs. In Experiment 3, males were allowed to copulate to ejaculation with an unscented female followed by exposure without access to a scented female. This treatment also supported the development of CEP. These results indicate that ejaculation plus a PEI are necessary for the development of CEPs and that the female must be present during the PEI for this to occur. These findings indicate that events during the PEI are the critical components of the UCS for CEP development.

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.

The nature of the conditioned response mediating olfactory conditioned ejaculatory preference in the male rat.

We have developed a model to study the influence of conditioning on sexual partner preference in the rat. In this model, pairing a neutral odor (almond) with copulation to ejaculation produces a subsequent preference to ejaculate with females bearing that odor. We refer to this phenomenon as a conditioned ejaculatory preference (CEP). The present study investigates the nature of the conditioned response that mediates CEP. Given equal mount and intromission distributions, but an unequal ejaculation distribution, we hypothesized that two mechanisms could account for CEP: facilitated ejaculation or selective ejaculation. To test these hypotheses, we examined the effect of omitting the olfactory conditioned stimulus (CS) or applying it to a nonreceptive female. Males trained with the CS paired with copulation (paired-trained males) failed to display evidence of delayed ejaculation when copulating with unscented females. Conversely, paired-trained males displayed CS-elicited copulatory behavior with CS-bearing nonreceptive females. In addition, we re-analyzed the data from the copulatory preference tests of previous experiments for CEP-displaying males. Again, we failed to find evidence of facilitated ejaculation with the scented female. However, the time between the last mount or intromission and ejaculation was increased if either occurred with the scented female. Furthermore, more mounts were directed towards the scented female near the end, but not at other points, of an ejaculatory series. These findings suggest that the paired-trained males attend to the scented female near the point of ejaculation, and are consistent with the hypothesis that CEP is mediated by selective ejaculation.

Basal forebrain damage and object-recognition in rats.

Damage to the basal forebrain (BF) produces permanent learning and memory impairments in humans. Most efforts to model these deficits in rats have focused on spatial memory dysfunction; this study was the first to assess the effects of BF damage in rats on the performance of a battery of object-memory tasks commonly employed to assess brain damage-produced amnesia in primates. The performance of rats with bilateral electrolytic lesions of the medial septum and diagonal band (MS/NDB) region of the BF was assessed on three object-memory tasks: nonrecurring items delayed nonmatching-to-sample (DNMS), simple object discrimination, and eight-pair concurrent object discrimination. Lesioned rats and sham-surgery controls were tested on the DNMS task at retention delays of 4, 15, 30, 60 and 120 s both before and after surgery. After surgery, the rats with MS/NDB lesions required significantly more trials than controls to relearn the nonmatching rule; and, once they relearned the rule, they were significantly and comparably impaired at all delays. This impairment did not diminish with either the passage of time or additional practice. In contrast, there were no significant differences between the MS/NDB-lesioned and control groups in the performance of either simple or concurrent object-discrimination tasks. The delay-independent nature of the DNMS deficit and the lack of deficits on the other two object-memory tasks suggest that the effect of the lesion is not the result of an impairment in retention.

Noncontingent drug exposure facilitates the development of contingent tolerance to the anticonvulsant effects of ethanol and diazepam in kindled rats.

Tolerance to anticonvulsant drug effects on kindled convulsions can result from drug exposure alone, but convulsive activity during drug exposure has a substantial facilitatory effect on tolerance development. Tolerance produced by drug exposure in the absence of a criterion response (in this case convulsions) has been termed pharmacologic tolerance (10); tolerance produced by drug exposure with concomitant performance of the criterion response has been termed contingent tolerance (1). The present study examines whether noncontingent drug exposure facilitates the development of contingent tolerance to the anticonvulsant effects of ethanol and diazepam. Amygdala-kindled, Long-Evans rats were treated with either ethanol (5.0 g/kg once daily for 21 days) or diazepam (5.0 mg/kg three times daily for 10 days) in the absence of convulsive stimulation to produce pharmacologic tolerance--control rats received treatments of vehicle. Then, all of the rats were rendered contingently tolerant by a series of "bidaily" (once every 2 days) injections (ethanol 2.0 g/kg or diazepam 2.0 mg/kg), each 1 h prior to a kindled convulsion. The rats that had received noncontingent exposure to ethanol or diazepam developed contingent tolerance significantly faster than the control rats. These results suggest that the mechanisms underlying pharmacologic and contingent tolerance to anticonvulsant drug effects are additive.

Persistence of the interictal emotionality produced by long-term amygdala kindling in rats.

Long-term amygdala kindling in rats results in large and reliable increases in emotional behaviour that model the interictal emotionality often observed in temporal lobe epileptics [Kalynchuk L. E. et al. (1997) Biol. Psychiat. 41, 438-451; Pinel J. P. J. et al. (1977) Science 197, 1088-1089]. These experiments investigated the persistence of these kindling-induced increases in emotional behaviour after the cessation of the kindling stimulations. In Experiment 1, rats received 99 amygdala or sham stimulations. Then, they were tested on three tests of emotionality (i.e. activity in an unfamiliar open field, resistance to capture from the open field, and activity in an elevated-plus maze) either one day, one week, or one month after the final stimulation. The rats tested one day after the last stimulation displayed substantial decreases in open-field activity, increases in resistance to capture and increases in open-arm activity on the elevated-plus maze; these effects decreased, but not to control levels, in the rats tested one month after the final stimulation. In Experiment 2, rats received 99 amygdala or sham stimulations, and their resistance to capture was assessed one day later. Then, after a 60-day stimulation-free period, the rats received another zero, one, 10, or 30 amygdala stimulations and their resistance to capture was reassessed one day later. The high levels of resistance to capture observed in the rats tested one day after the 99 stimulations declined significantly during the 60-day stimulation-free period, but it remained significantly above control levels. However, the administration of 30 additional stimulations reinstated asymptotic levels of resistance to capture. These results provide the first systematic evidence that kindling-induced increases in emotional behaviour persist at significant levels for at least two months following the termination of kindling stimulations. Thus, they suggest that the neural changes underlying the genesis of interictal emotionality may be closely related to those mediating epileptogenesis itself.

Contingent tolerance, compensatory responses, and physical dependence in diazepam-treated amygdala-kindled rats.

Three groups of amygdala-kindled rats received 10 bidaily treatment trials: On each trial, the drug-before group received a diazepam (2.5 mg/kg i.p.) injection 1 hr before a convulsive stimulation, the drug-after group received a diazepam injection 1 hr after a stimulation, and the vehicle control group received a vehicle injection either 1 hr before or 1 hr after a stimulation. After treatment, only the drug-before group displayed significantly longer forelimb clonus under the influence of diazepam (that is, they displayed contingent tolerance to diazepam's anticonvulsant effect) and significantly longer forelimb clonus while drug free. Following a 14-day retention period, the rats in the drug-before group retained significant levels of contingent tolerance but did not display significant increases when tested drug free. These data suggest that compensatory responses do not play a causal role in the expression of contingent tolerance.

Changes in emotional behavior produced by long-term amygdala kindling in rats.

The effects of long-term amygdala kindling on emotional behavior were investigated. In Experiment 1, rats received 99 basolateral amygdala, central amygdala, or sham stimulations. The rats in both kindled groups displayed more resistance to capture from an open field and more open-arm activity on an elevated plus maze than did the sham control rats. In Experiment 2, rats received either 20, 60, or 100 amygdala stimulations or sham stimulations. Compared to the sham controls, the kindled rats explored less during the first 30s in a novel open field, avoided the central area of the open field, resisted being captured from the open field, and engaged in more open-arm activity on the elevated plus maze. The magnitude of these effects was greatest in the 100-stim rats and least in the 20-stim rats. Together, these results suggest that long-term amygdala kindling in rats is a useful model for studying the emotionality associated with temporal lobe epilepsy.

Tolerance to the anticonvulsant and ataxic effects of pentobarbital: effect of an ascending-dose regimen.

We assessed the effect of an ascending-dose regimen on the development of tolerance to the anticonvulsant and ataxic effects of pentobarbital in four groups of amygdala-kindled rats. Each rat received 20 bidaily (one every 48 h) trials in which an intraperitoneal (IP) pentobarbital or vehicle injection was delivered 1 h before a convulsive amygdala stimulation. On each trial, the rats in the three pentobarbital groups received either a high dose (50 mg/kg), a low dose (10mg/kg), or ascending doses of pentobarbital that began at 10 mg/kg and increased to as high as 26 mg/kg by 1 mg/kg increments as tolerance developed to its anticonvulsant effect; the rats in the vehicle group received saline. The rats in the ascending-dose condition displayed significantly more tolerance to the anticonvulsant effect of pentobarbital than did the other rats; in contrast, the high-dose rats displayed more tolerance to the ataxic effect of pentobarbital than did the other rats. These findings extend previous reports of the facilitatory effect of ascending-dose regimens on the development of tolerance to the anticonvulsant effect of benzodiazepines, and show that the facilitatory effect of ascending-dose regimens does not extend to all drug effects.

Contingent drug tolerance: differential tolerance to the anticonvulsant, hypothermic, and ataxic effects of ethanol.

The kindled-convulsion model of epilepsy was used to study contingent tolerance to ethanol's (1.5 g/kg; IP) anticonvulsant, hypothermic, and ataxic effects in adult male rats. In the present experiments, three groups of amygdala-kindled rats received a series of bidaily (one every 48 h) convulsive stimulations: one group received ethanol 1 h before each stimulation; one group received ethanol 1 h after each stimulation; and another group served as the saline control. Tolerance to ethanol's anticonvulsant effect (Experiments 1 and 2) was greatest in those rats that received ethanol before each convulsive stimulation; whereas, tolerance to ethanol's hypothermic (Experiments 1 and 2) and ataxic (Experiments 2) effects developed in both groups that received ethanol. These results were predicted on the basis of the drug-effect theory of drug tolerance: the theory that functional drug tolerance is an adaptation to the disruptive effects of drugs on concurrent patterns of neural activity, not to drug exposure per se.

Dissipation of contingent tolerance to the anticonvulsant effect of diazepam: effect of the criterion response.

The effect of convulsive stimulations on the dissipation of tolerance to the anticonvulsant effect of diazepam was investigated using the kindled-convulsion model. Amygdala-kindled rats were rendered tolerant to diazepam's anticonvulsant effect by 25 "bidaily" (one/48 h) diazepam injections (2.5 mg/kg), each followed 1 h later by a convulsive stimulation. They were then divided into nine groups for the tolerance-dissipation phase of the experiment. Of the nine groups, three received bidaily control handling for one trial, three trials, or seven trials; three received bidaily saline injections, each 1 h before a convulsive stimulation, for one, three, or seven trials; and three received bidaily diazepam injections, each 1 h after a convulsive stimulation, for one, three, or seven trials. Finally, each rat received a tolerance-retention test (i.e., a diazepam injection followed 1 h later by a convulsive stimulation) 48 h after its last tolerance-dissipation trial. The tolerance dissipated gradually but completely over the 4-, 8-, and 16-day test intervals in the rats that received a convulsive stimulation before each injection during the tolerance-dissipation phase, whether they were injected with saline or diazepam; in contrast, tolerance did not dissipate in the rats that received saline injections but no stimulations. Remarkably, the discontinuance of the bidaily diazepam injections, even for 16 days, was not sufficient to dissipate the tolerance that had developed to diazepam's anticonvulsant effect; nor was the continuation of the bidaily diazepam injections sufficient to keep tolerance from dissipating.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of an ascending dose regimen on the development of tolerance to the anticonvulsant effect of diazepam.

The effect of an ascending dose regimen on the development of tolerance to diazepam's anticonvulsant effect was assessed. During the 22 trials of the tolerance development phase, amygdala-kindled rats received either a series of dosage injections ranging from high (10 mg/kg), to low (1.0 mg/kg), and ascending (1.0 mg/kg and increased by 0.2-mg/kg increments to 3.0 mg/kg) or saline injections. Diazepam was administered by ip injection once every 48 hr, and each injection was followed 1 hr later by a convulsive stimulation. The ascending dose rats displayed significantly more tolerance to the anticonvulsant effect of diazepam than did the high dose, low dose, or saline rats. By contrast, both the ascending and high dose rats displayed a significant withdrawal effect (i.e., increased duration of convulsions) after the cessation of diazepam injections. Results demonstrate that administration of ascending dosages can facilitate the development of tolerance to anticonvulsant drug effects and that tolerance and withdrawal are not necessarily inextricably related.