Selected opioids and responding for intracranial reinforcement.

Rats fixed with chronically indwelling electrodes for intracranial stimulation (ICS) of the lateral hypothalamus pressed a lever during daily sessions for a fixed intensity of ICS. Before some sessions, they were given placebo, or ethylketocyclazocine (EKC) in racemic or isomeric forms [either (+)EKC or (-)EKC]. Naloxone (NX) was also given with the agents. The racemate facilitated pressing across a narrow range of small doses (centered about 0.02 mg/kg). At no dose did (-)EKC, a potent analgesic, facilitate pressing and typically depressed it. (+)EKC, at doses of 0.04 and 0.08 mg/kg, facilited pressing. These data provide further confirmation that opioid analgesia and ability to enhance pressing are separable. When NX was given with a large dose of the racemate, paradoxically pressing for ICS was facilitated. Apparently, NX selectively blocked the effects of (-)EKC. SKF 10047 was also administered in racemic and isomeric forms. All three forms produced some facilitation of pressing at small doses (e.g., 0.75 mg/kg) and depressed pressing at large doses (e.g., 5.0 mg/kg).

alpha(2)-Adrenoceptor antagonists and propensity to take alcoholic beverages.

Rats were maintained on a daily regimen involving a 2h opportunity to take both water and a sweetened alcoholic beverage (12% ethanol, 0.25% saccharin). After 3 weeks on this regimen, rats regularly take substantial amounts of alcohol. After stabilization, injections of alpha(2)-adrenergic antagonists were administered, 15min before the opportunity to drink. Yohimbine and methoxyidazoxan dose relatedly decreased intake of alcoholic beverage and increased intake of water. In Experiment 2, a number of rats were taken off the daily regimen for 9 days, then returned to it. Across the first 12 days of the reinstated daily regimen, half the rats received placebo and half methoxyidazoxan. The group receiving placebo rapidly returned to taking large amounts of alcoholic beverage while the group receiving methoxyidazoxan did not. In Experiment 3, it was shown that a dose of methoxyidazoxan that decreased intakes of alcoholic beverage did not decrease intakes of other palatable beverages. In Experiment 4, it was shown that yohimbine persistently reduced intakes of alcoholic beverage with daily administration. These results indicate that alpha(2)-antagonists might be effective pharmaceutical adjuncts to other treatments for alcohol abuse and alcoholism.

Isradipine in combination with naltrexone as a medicine for treating cocaine abuse.

Rats were fixed with chronically indwelling electrodes for intracranial stimulation (ICS) of the medial forebrain bundle of the lateral hypothalamus. They were trained to press a bar in a Skinner box for the ICS. After stable rates of pressing for both a low and a high intensity of ICS were achieved during daily sessions, the rats were given doses of cocaine before the daily sessions. Cocaine produced its characteristic effect of enhancing rates of pressing. With continuance of daily sessions under the influence of cocaine, the rats received daily for 5 days a combination of isradipine and naltrexone. Doses of isradipine and naltrexone were smaller than a dose of either one that might modify pressing. The combination of isradipine and naltrexone blocked cocaine's enhancement of pressing for ICS. The same combination of isradipine and naltrexone did not reduce rates of pressing for ICS when cocaine was not given. These results indicate that a combination of isradipine and naltrexone is apt to be an effective pharmacological adjunct to other treatments for cocaine abuse.

Naltrindole, an opioid delta receptor antagonist, blocks cocaine-induced facilitation of responding for rewarding brain stimulation.

Recent experimental results have led to the suggestion that opioid antagonists can modulate the reinforcing properties of cocaine. In this experiment, rats were fixed with chronically indwelling bipolar electrodes for stimulation of the medial forebrain bundle (MFB) as it courses through the hypothalamus. Rats were taught to press a lever for brief trains of electrical stimulation of the MFB. Subsequently, they were allowed to press for varying intensities of stimulation daily until their response rates were stable. Cocaine (5 mg/kg, s.c.) enhanced the rate of pressing for lower intensities of brain stimulation. Naltrindole (3 mg/kg, i.p.) had no effect on response rate alone but blocked the cocaine-induced facilitation of pressing for rewarding brain stimulation. An implication that can be drawn from these data is that naltrindole, or other delta-selective opioid antagonists, might be effective as medicines for use in treating cocaine abuse.

Opioidergic manipulations affect intake of 3% NaCl in sodium-deficient rats.

On six weekly occasions, a 3% NaCl solution was presented along with water to rats for 2 h 1 day after being treated with furosemide, a diuretic/natriuretic drug that causes a strong hunger for 3% NaCl. On some of the days, the sodium-hungry rats were injected with morphine in doses ranging from 0.3 to 10.0 mg/kg. Morphine produced biphasic effects on intake of 3% NaCl, with doses of 0.3-3.0 mg/kg increasing intakes dose dependently and 10.0 mg/kg decreasing intakes. The 3.0-mg/kg dose nearly doubled rats' mean intake of 3% NaCl. In contrast, naltrexone, an opioid receptor antagonist, reduced intake of 3% NaCl about 25-40% across doses ranging from 0.1 to 10.0 mg/kg. At some doses of morphine and naltrexone, NaCl ingestion was affected without significant influence of water intake. Therefore, it can be inferred that endogenous opioidergic systems participate in the control of NaCl drinking by sodium-deficient rats. The range of demonstrations of opioid involvement in the control of ingestion can now be extended to the hunger for hypertonic NaCl induced by sodium depletion.

A combination of isradipine and naltrexone blocks cocaine's enhancement of a cocaine place preference.

Rats were conditioned by pairing cocaine with one side of an alley and placebo with the other. After conditioning, compared to Baseline and a placebo-control group, rats spent more time in the place of cocaine experience. Subsequently, there were further tests except now cocaine was given just before the test session in addition to one of two other kinds of injections. One of these additional injections was a placebo and the other was a combination of a small dose of isradipine (1 mg/kg) and a dose of naltrexone (3 mg/kg) (ISR + NTX). Measures of gross activity (movement from one side of the alley to the other) were taken during testing. ISR + NTX blocked cocaine's ability to sustain a place preference. ISR + NTX also blocked sensitization of cocaine's ability to enhance locomotor activity. This blockade of cocaine's usual effects indicates that ISR + NTX may have a role in treating cocaine use disorders.

PCP, THC, ethanol, and morphine and consumption of palatable solutions.

Water-deprived rats were given daily opportunities (2.0-hr sessions) to take water or a sweet solution (20% or 24% sugar-water). After stable intakes of each fluid were achieved, the effects of phencyclidine hydrochloride (PCP), delta-9-tetrahydrocannabinol (THC), ethanol (E), and morphine (M) on intakes were tested. PCP, THC, and M all enhanced intake of the sweet solution, while E produced varying effects across doses tested. With other rats, nearly the same procedure was used except that the test solution presented with water was 0.9% sodium chloride. Doses of PCP enhanced intake of the salty solution. These data, combined with the data from similar studies of the effects of opioids and benzodiazepines, indicate that a wide variety of agents that are self-administered also modify intake of ingesta.

Naltrindole, a delta-opioid antagonist, blocks MDMA's ability to enhance pressing for rewarding brain stimulation.

Twelve rats were each fixed with a chronically indwelling bipolar electrode for stimulation of the medial forebrain bundle as it courses through the hypothalamus. These rats were trained to press a bar for intracranial stimulation of 0.3-s trains of 60 Hz sine waves for 10 min daily at three intensities. One intensity was just above threshold for maintaining pressing, one intensity was a high intensity that sustained considerable pressing, but not maximum pressing, and the other was intermediate to the others. After stable rates of pressing were obtained, rats received MDMA daily. MDMA significantly increased rates of pressing. Prior to a day when rats received MDMA, they also received an injection of naltrindole, a selective delta-opioid receptor antagonist. Naltrindole blocked MDMA's enhancement of pressing for reinforcing brain stimulation.

Isradipine blocks cocaine's ability to facilitate pressing for intracranial stimulation.

Using rats pressing for rewarding electrical intracranial stimulation of the medial forebrain bundle, it was found that a single administration of isradipine blocked the rate-enhancing effects of cocaine (5.0 mg/kg) at doses of 3.0 and 10.0 mg/kg. Also, when isradipine (3.0 mg/kg) was administered alone (without cocaine) for 5 consecutive days, pressing for intracranial stimulation was not reduced relative to placebo levels. In another experiment, isradipine (3.0 mg/kg) persistently blocked the rate-enhancing effects of cocaine (5.0 mg/kg) across 5 consecutive days. These results support the continued investigation of isradipine as a useful adjunct to other treatments for cocaine addiction.

MDMA produces a conditioned place preference and elicits ejaculation in male rats: a modulatory role for the endogenous opioids.

Methylenedioxymethamphetamine (MDMA) can produce a conditioned place preference (CPP) among rats. The ability of MDMA to produce a CPP was assessed while some rats were under the influence of naltrexone, 56 mg/kg, given 4 h before conditioning. Naltrexone attenuated MDMA's ability to produce a CPP without completely blocking MDMA's effects. Having noticed previously the production of seminal plugs by rats receiving MDMA, the presence of seminal plugs was recorded across the 8 days of conditioning. Roughly half of the rats receiving 6.3 mg/kg of MDMA left plugs during the conditioning period, while over two-thirds of those receiving a combination of MDMA and naltrexone left plugs. A second study, assessing further doses of MDMA, tabulated the drug's effects on the production of seminal plugs across 3 h. Besides eliciting ejaculation, MDMA also led to increased urination and defecation and a loss of body weight. These results support suggestions that the endogenous opioids modulate the reinforcing properties of stimulant drugs and affect male sexuality.

Methylenedioxymethamphetamine's capacity to establish place preferences and modify intake of an alcoholic beverage.

Doses of 0.2, 2.0, 6.3 and 20.0 mg/kg 3,4-methylenedioxymethamphetamine (MDMA), a putative neurotoxin at serotonergic neurons and a recreational drug, were assessed using Sprague-Dawley rats in the conditioned place preference (CPP) test. Also, the drug's effects on intake of a sweetened ethanol solution (ES) was assessed. The CP testing involved multiple administrations of MDMA with frequent periodic testing (weekly for 4 weeks) of MDMA's effects. Doses of 2.0 and 6.3 mg/kg produced positive CPPs with every test. MDMA also affected rats' gain in body weight across the 4 weeks of dosing. The 2.0 mg/kg reliably incremented gain in body weight, while the 20.0 mg/kg dose reliably attenuated it. In the drinking experiment, water-deprived rats (22 h/day) were given daily opportunities to drink either tap water or a sweetened ES. When stable intakes were achieved, MDMA's effects were assessed across repeated daily administrations (12 days) and subsequently (16 days). MDMA, dose-relatedly, decreased intake of both ES and water with the highest dose leading to marked loss in body weight. Intakes of fluids were not modified markedly subsequent to dosing. In summary, MDMA is an agent that produces a positive CPP (providing further evidence for MDMA's abuse liability), produces changes in weight gain and nonselectively reduces fluid intake among fluid-deprived rats.

Measuring morphine's capacity to establish a place preference.

A series of experiments are described providing an assessment of the procedures of conditioned place preference (CPP) testing involving an automated system having 12 separate chambers. Experiment 1 provides data to demonstrate (a) that in these chambers no initial preferences for one side over the other exists among rats, (b) that this neutrality of sides is not affected by session lengths between 15 and 60 min, and (c) that the optimal session length for tests in these chambers is on the order of 30 min. Experiment 2 demonstrates the stability of control groups' scores across a number of conditioning and testing sessions. Experiments 3 and 4 provide data to demonstrate (a) that a positive CPP can be established in our chambers using injections of morphine, (b) that a regimen of dosing with unequal numbers of days of putative and alternate conditioning is a reliable and conservative test of the opioid's ability to establish a CPP, and (c) that although the activity of rats decreases across a session, the general activity of rats before and after conditioning procedures is the same. Experiment 5 replicates the procedures employed by Scoles and Siegel (25) and demonstrates that the tendency for rats to explore novel environments is strong, and care must be taken to provide an opportunity for rats to pair different experiences with each side of the chamber in order for a CPP to emerge.

Toward understanding ethanol's capacity to be reinforcing: a conditioned place preference following injections of ethanol.

Rats that had previously consumed a 6% ethanol (ETOH) solution daily for 26 days and rats without such a history served as subjects in a test for the ability of ETOH to establish a conditioned place preference. The time of putative conditioning was from 4 to 8 min after injections of ETOH, 1 g/kg. The combination of programming the period of putative conditioning to be shortly after injections and using rats habituated to drinking ETOH allowed a conditioned place preference to emerge after only a few conditioning trials. Such a result potentially reveals features of the way ETOH achieves its reinforcing capability and sets the stage for understanding the mechanism of that reinforcement.

Isradipine combined with naltrexone persistently reduces the reward-relevant effects of cocaine and alcohol.

Previous studies have revealed that the combination of small doses of isradipine and naltrexone (ISR&NTX) blocks the ability of cocaine to enhance pressing for rewarding, lateral hypothalamic brain stimulation. Further, such combinations also reduce rats' intakes of alcoholic beverages. Here, we asked whether ISR&NTX would lose its ability to reduce the reinforcing effects of cocaine and alcohol when given daily. Specifically, after almost 2 months of daily injections, ISR&NTX blocked the expression of a cocaine-induced conditioned place preference (CPP). By themselves, ISR and NTX were not effective at blocking cocaine's effects. Subsequent to the CPP procedures, the rats continued to receive daily injections for another 3 weeks. During this time, they were given access to water and an alcoholic beverage for 2 h a day. As expected, placebo controls gradually increased their daily intakes until they were taking about 2 g/kg of ethanol daily. ISR, NTX, and ISR&NTX blocked the typical pattern of intakes. At the end of the 3-week period, the rats had received 80 consecutive daily injections. The data suggest that the salient effects of ISR&NTX do not wane. The data support the idea that ISR&NTX would be a useful pharmacotherapy for poly drug abuse.

Responding for rewarding brain stimulation: cocaine and isradipine plus naltrexone.

Rats, fixed with chronically indwelling electrodes for electrical intracranial stimulation (ICS) of the lateral hypothalamus, were taught to press a bar for ICS. Once pressing rates became stable, during daily 20-min sessions, rats were given cocaine (5 or 20 mg/kg) before the sessions. When given daily, cocaine consistently enhanced rates of pressing. When a combination of small doses of isradipine (e.g., 1 mg/kg) and naltrexone (3 mg/kg) were given before cocaine administration. the combination blocked cocaine's enhancement of pressing for ICS. The combination, however, neither reduced rates of pressing below those observed under placebos (i.e., baseline conditions) nor reduced rates when no cocaine was given. Naltrexone and isradipine (in the dose used in the combination) by themselves did not block cocaine's effects. This profile of effects indicates that a combination of isradipine and naltrexone is apt to be useful in treating cocaine use disorders.

Amlodipine, a calcium channel inhibitor, and cocaine and ethanol's reinforcing effects.

The effects of amlodipine (from 0.1 to 3.0 mg/kg) on rats' pressing for rewarding brain stimulation, with and without cocaine administration, were assessed. None of the doses reliably modified the effects of cocaine. Also, amlodipine was given to two groups of rats taking alcohol: one group that was regularly taking a sweetened alcoholic beverage and the other taking an unsweetened alcoholic beverage. The only discernible effects of amlodipine on alcohol intake were associated with the highest dose and only with rats taking the sweetened beverage. The effects of this high dose could easily be attributable to behavioral toxicity elicited by the dose. In contrast, and confirming previous work, isradipine, another calcium channel inhibitor, produced reliable reductions on both cocaine's and alcohol's reinforcing effects. Despite the similarity of isradipine and amlodipine, isradipine apparently has some unique features with respect to cocaine and alcohol.

Daily water deprivation potentiates ethanol intake beyond a period of water intake.

Rats were given a daily opportunity to drink sweetened ethanol solution or water after 22 hr of deprivation of water. Their intakes of the ethanol solution increased gradually across three weeks and then remained relatively stable. These procedures examined the effects of having an opportunity to drink water for 30 min, before the opportunity to drink ethanol solution or more water. Even though 30 min of opportunity to drink water lessened intake of further water, this experience did not lessen rats' intake of the subsequently presented ethanol solution. The complex events of the daily cycle of deprivation and limited opportunities to ingest, which potentiate intake of flavored alcoholic beverage, are not modified substantially by merely providing water.

Persistence and specificity of small doses of morphine on intake of alcoholic beverages.

Subsequent to water deprivation, male rats were given daily, 1.5-hr opportunities to take either water or a sweetened ethanol solution (ES). Each day, 15 min before the session, rats received a subcutaneous injection of either morphine (1.0 mg/kg) or saline. Across daily sessions, rats given saline gradually increased their intake of ES, until they were eventually taking about 2.0 to 3.0 g of ethanol/kg a session. Rats receiving morphine took greater amounts of ES from nearly the first opportunities. Additional tests assessed the effects of small doses of morphine on intakes of some sucrose solutions, and sweetened solutions containing methanol or propanol. The data support the conclusion that small doses of morphine persistently increase intake of ES across many days (up to 100) of testing, but that the effect is not unique to ES. Even though morphine's effects are not specific to ethanol, the fact that morphine persistently increases intake of ES is of interest with respect to theories of alcoholism.

Morphine increases intake of beer among rats.

Rats were maintained on a daily regimen of 22 h of water deprivation followed by a 2-h opportunity to take water and sweetened alcoholic beverage containing 12% ethanol. After 30 days, the alcoholic beverage was changed to beer containing either 3% or 6% ethanol. After 20 daily sessions with beer, they received, before the next session, an injection of saline. On the next day, they received a 1.0 mg/kg injection of morphine before the session. Morphine reliably increased rats' mean intake of both kinds of beer. Subsequently, the concentration of ethanol in each groups' beer was changed. The 3% group's beer was switched to 6%, and the 6% group's to 3%. Both groups altered their mean intake of beer in an apparent attempt to maintain intakes of nearly the same amount of ethanol, but presentation of 6% beer resulted in greater intakes of ethanol.

Constant infusions of morphine and intakes of sweetened ethanol solution among rats.

Previous studies have indicated that injections of small doses of morphine increase rats' intake of solutions containing ethanol when rats have a choice of either water or a solution containing ethanol. In this experiment, rats which were implanted with osmotic pumps that delivered constant infusions of morphine (0.6 mg/kg/hr across 24 days) had elevated daily intakes of ethanol, as compared to controls, from the second day of opportunity to take the alcoholic beverage until the pumps were removed. In addition, half of the rats with pumps infusing morphine also received injections of morphine (1.0 mg/kg) just before the 1.5-hr opportunity to take alcoholic beverage or water every day for 8 days. Across this 8-day period, these rats took a mean of 5.18 g of pure ethanol/kg of body weight (g/kg) during the 1.5-hr opportunity to take the alcoholic beverage. This was reliably more than the mean of 4.02 g/kg that their counterparts (having morphine pumps and receiving injections of saline) took across the same period. These data support the hypothesis that a surfeit of opioidergic ligand may potentiate drinking of alcoholic beverages.