Yavin Shaham

Stress reinstates cocaine-seeking behavior after prolonged extinction and a drug-free period

Abstract We have shown previously, using an animal model of relapse, that acute exposure to intermittent footshock stress induces reinstatement of heroin-taking behavior in rats. Here we report that in rats trained to self-administer cocaine, exposure to acute intermittent footshock stress induces reinstatement of cocaine-taking behavior after prolonged extinction sessions and after a 4- to 6-week drug-free period; an effect comparable to that induced by a priming injection of cocaine. Animals were initially allowed to self-administer cocaine HCl (1.0 mg/kg per infusion, IV) during one 3-h session/day for 12 days. Subsequently, extinction conditions were introduced by substituting saline for cocaine so that lever-pressing resulted in IV infusions of saline rather than of drug. Extinction conditions were maintained until animals made 15 responses or less in the 3 h, after which animals were given saline infusions at the start of each daily session to establish baseline responding of ten responses or less. Subsequently, animals were tested for reinstatement of responding for saline infusions following a non-contingent injection of cocaine (2.0 mg/kg, IV) and exposure to intermittent footshock (10 min, 0.5 mA, 0.5 s on, mean off period of 40 sec). After an additional 4- to 6-week drug-free period, tests for reinstatement were repeated. Reinstatement of cocaine-taking behavior was observed in both sets of tests in response to footshock and cocaine. These results extend previous reports from this laboratory that footshock stress is an effective stimulus for reinstatement of drug-taking behavior in the rat.

Stress reinstates heroin-seeking in drug-free animals: An effect mimicking heroin, not withdrawal

Exposure to 10 min of footshock stress (1 mA; 0.5 s on, with a mean off period of 40 s) reinstated heroin-seeking behavior in heroin-experienced, drug-free rats after many sessions of extinction and up to 6 weeks after last exposure to heroin. In reinstating the behavior, the footshock mimicked the effect of a non-contingent priming infusion of heroin (50 µg/kg). By contrast, the aversive state of acute opioid withdrawal induced by injection of the opioid receptor antagonist naltrexone (5 mg/kg, SC), following an acute injection of morphine (10 mg/kg, SC), had no effect on heroin-seeking behavior. In a second experiment it was shown in drug naive animals that these parameters of footshock increased dopamine overflow in the nucleus accumbens, a terminal region of the mesolimbic dopamine system implicated in the reinforcing effects of drugs. Similarly, dopamine overflow was increased by an injection of 10 mg/kg morphine, SC, an effect that was reversed by an injection of 5 mg/kg naltrexone given 40 min after to induce the withdrawal condition. A possible interpretation of the present results is that stressors can reinstate drug-taking behavior by activating neural systems in common with those activated by heroin.

Serotonin Neurotoxicity after (±)3,4-Methylenedioxymethamphetamine (MDMA; “Ecstasy”): A Controlled Study in Humans

(±)3,4-Methylenedioxymethamphetamine (MDMA; “Ecstasy”), an increasingly popular recreational drug, is known to damage brain serotonin 5-hydroxytryptamine (5-HT) neurons in experimental animals. Whether MDMA is neurotoxic in humans has not been established. Thirty MDMA users and 28 controls were admitted to a controlled inpatient setting for measurement of biologic and behavioral indexes of central 5-HT function. Outcome measures obtained after at least 2 weeks of drug abstinence included concentrations of monoamine metabolites in cerebrospinal fluid (CSF), prolactin responses to L-tryptophan, nociceptive responses to ischemic pain, and personality characteristics in which 5-HT has been implicated (i.e., impulsivity and aggression). Subjects with a history of MDMA exposure had lower levels of CSF 5-hydroxyindoleacetic acid (the major metabolite of 5-HT) than controls (p = .002). Although they resembled controls in their prolactin response to L-tryptophan and their response to ischemic pain, MDMA users had lower scores on personality measures of impulsivity (p = . 004) and indirect hostility (p = .009). The CSF findings suggest that 5-HT neurotoxicity may be a potential complication of MDMA use. Further, differences in personality support the view that 5-HT systems are involved in modulating impulsive and aggressive personality traits. Additional studies of MDMA-exposed individuals are needed to confirm and extend the present findings. Such studies could help elucidate the role of 5-HT in normal brain function as well as in neuropsychiatric disease states.

Effects of opioid and dopamine receptor antagonists on relapse induced by stress and re-exposure to heroin in rats

The effects of blockade of opioid and dopamine receptors on relapse to heroin-seeking induced by footshock stress and re-exposure to heroin were examined in a reinstatement procedure. Male rats were trained to self-administer heroin (100 µg/kg per infusion, IV; four 3-h sessions/day for 8–11 consecutive days). Extinction sessions were given for 5–7 days during which saline was substituted for heroin. In nine groups, the effects on relapse induced by footshock (10 min, 0.5 mA, 0.5 s on with a mean off period of 40 s), heroin priming (0.25 mg/kg), and saline priming were studied after pretreatment with either naltrexone (1 or 10 mg/kg, SC), the D1-like receptor antagonist SCH 23390 (0.05 or 0.1 mg/kg, IP), the D2-like receptor antagonist raclopride (0.25 or 0.5 mg/kg, IP), the mixed dopamine antagonist flupenthixol decanoate (3 or 6 mg/kg, IM), or IP injection of saline (control condition). Naltrexone, flupenthixol, raclopride, and the highest dose of SCH 23390 attenuated heroin-induced relapse: only the mixed DA receptor antagonist, flupenthixol, attenuated footshock-induced relapse. These results, and those from microdialysis showing that heroin elicits greater locomotor activity and DA release in the nucleus accumbens than footshock, suggest that the neurochemical events underlying stress- and heroin-induced relapse are not identical.

Exposure to mild stress enhances the reinforcing efficacy of intravenous heroin self-administration in rats

The effect of a mild footshock on intravenous heroin self-administration was examined in male rats. Animals in the stress condition were exposed to 10 min of intermittent footshock (0.5 mA; 0.5 s on, with a mean off period of 40 s) before each of four daily self-administration sessions. Animals in the control group were not exposed to footshock. Following acquisition of heroin-reinforced behavior (100 µg/kg per infusion), during which no group differences emerged, animals were placed on a progressive ratio schedule of reinforcement and were subsequently tested under a decreasing series of doses. Animals exposed to footshock before each drug session had higher rates of lever pressing for heroin and achieved higher final ratios on the progressive ratio schedule than animals in the control group at the higher doses of heroin. Thus, under the conditions of this experiment, exposture to mild intermittent stress appeared to enhance the reinforcing efficacy of heroin. The parameters of footshock used in the present study, and its relation to drug availability may characterize conditions under which stress leads to increased opioid abuse.

Immobilization stress-induced oral opioid self-administration and withdrawal in rats: role of conditioning factors and the effect of stress on “relapse” to opioid drugs

The effect of 15 min/day of immobilization (IM) stress on oral self-administration (SA) of morphine (0.5 mg/ml) or fentanyl (25 µg/ml) and withdrawal was examined in rats. In addition, the role of conditioning factors in these effects was assessed. For each drug, four groups of subjects were exposed for 50 days to IM stress prior to the drug SA period [Paired-Stress (P-S) groups], to IM stress prior to the drug SA period on half of the days and after the drug SA period on the rest of the days [Partial Paired-Stress (PP-S) groups], to IM stress several hours after the drug SA period [Unpaired-Stress (UP-S) groups], or to no IM stress [Control (C) groups]. The P-S and PP-S groups increased their drug SA during choice days in which both the opioid solution and water were available, and tended to manifest a more severe withdrawal syndrome after a naloxone challenge compared with the UP-S and C groups. Reinstatement of the opioid SA under conditions of paired-stress or no stress was further examined after 3 weeks without exposure to either stress or drugs. The paired stress animals had higher levels of drug SA and manifested a more severe withdrawal syndrome than those tested without stress. These results indicate that the learned association between exposure to stress and the drug availability may mediate, in part, the stress-induced enhancement of opioid SA and withdrawal effects.

Effect of stress on oral morphine and fentanyl self-administration in rats

The effect of immobilization stress (15 min/day) or no stress on oral morphine (0.25-0.5 mg/ml) or fentanyl (5-20 micrograms/ml) self-administration was examined in rats. Animals had access to a morphine or fentanyl solution for 4 days, followed by a single-choice day of access to the opioid solution and a separate water bottle. This 5-day cycle was repeated five times for 7 h/day in home cages. Morphine consumption and preference were assessed for an additional 30 days (i.e., six more cycles) in a subgroup of subjects. Plasma corticosterone levels in the stress groups indicated that the stress manipulation was effective. Over the course of the experiment, animals in the stress groups significantly increased their preference for the opioid solutions during choice days compared to nonstress controls. Morphine preference after 55 days was twice as high in the stress group (70% morphine/30% water) in comparison to controls (34% morphine/66% water). These results indicate that stress increases oral opioid self-administration in rats. Future directions and the implications of this work are discussed.

Ovarian hormones do not affect the initiation and maintenance of intravenous self-administration of heroin in the female rat

The effect of ovarian hormones on the initiation and maintenance of heroin intravenous self-administration was studied in ovariectomized female rats. In Experiment 1 (initiation), the behavior of groups of females (n = 8) ovariectomized (OVX), injected with 10 µg estradiol benzoate (EB) every 3 days (OVX+EB), and intact, was compared with that of intact males (n = 7) exposed to an ascending series of doses of heroin (6.125–50 µg/kg per infusion) on a fixed ratio—FR-1—reinforcement schedule. Twelve 3-h sessions per dose were given, 2 per day in the dark and 2 in the light period of a 12:12-h light:dark cycle. No differences in rate of responding between groups were observed at any of the heroin doses during acquisition or during tests given after acquisition on a descending series of doses. In Experiment 2 (maintenance), 7 OVX female rats were trained to self-administer heroin on an FR-1 reinforcement schedule. They were then switched to a progressive ratio schedule, 4 sessions per day, 4 h each, and a regimen of EB and progesterone (P) injections was initiated during which all animals were tested over a 5-day period (cycle) at each of four doses of heroin (50, 25, 12.5, and 0.0 µg/kg per infusion). Neither EB, given on Day 3, nor P, given on Day 5 of each cycle, affected the highest ratio achieved (breakpoint) or number of heroin infusions taken. These results suggest that the sensitivity of female and male rats to the reinforcing effects of heroin is not different, and that, in females, the reinforcing effects are not affected by circulating ovarian hormones.

Temporal factors in the effect of restraint stress on morphine-induced behavioral sensitization in the rat

The role of associative factors in the effect of 15 min/day of restraint stress on morphine-induced behavioral sensitization was examined. Male rats were initially given seven systemic (10 µg/kg, IP) or intraventral tegmental area (VTA, 5 mg/side) injections of morphine, and were exposed to restraint, either just prior to drug injection (Paired-Stress) or 24 h after injection (Unpaired-Stress), or to no restraint (Control). In subsequent tests for behavioral sensitization to low doses of morphine (0.75 or 3.0mg/kg, IP), animals in the Paired-Stress condition were more active than animals in the Unpaired-Stress or Control conditions. These results indicate that temporal and possibly associative factors may contribute to stress-induced changes in sensitization to the behavioral activating effects of opioids.

Effects of catecholamine depletion on alertness and mood in rested and sleep deprived normal volunteers

Alpha-methyl-para-tyrosine (AMPT), a tyrosine hydroxylase inhibitor, was used to evaluate the physiologic role of central nervous system catecholamines at modulating alertness and mood. Forty healthy males were randomized to one of four conditions: AMPT in a rested condition; AMPT plus 40.5 hours of total sleep deprivation; placebo plus sleep deprivation; or placebo in a rested condition. Repeated measures of alertness and mood revealed that treatment with AMPT or sleep deprivation increased sleepiness, and combined treatment produced greater sleepiness than either treatment alone. In contrast, although combined treatment with AMPT and sleep deprivation led to large increases in negative mood, neither treatment alone produced consistent mood changes. These findings are consistent with the view that sleep deprivation is associated with decreased functional catecholamine neurotransmission. Furthermore, mood effects following sleep deprivation plus AMPT suggest that catecholamines may be involved in mood changes during sleep deprivation.