Opioid and nonopioid conditional analgesia: the role of spinal opioid, noradrenergic, and serotonergic systems.

The first aim of this study was to test whether opioid mediation of conditional analgesia is inversely related to the relative amount of training that rats received, a prediction derived from the severity hypothesis. Naltrexone attenuated the analgesia among subjects in the low-training group but had no effect in rats given an extended amount of training. The second aim of this study was to investigate whether noradrenergic, opiate, and serotonergic spinal systems play a role in conditional analgesia. Intrathecal administration of yohimbine, an alpha 2 antagonist, completely blocked both opioid and nonopioid forms of conditional analgesia. Spinal administration of either quaternary naltrexone or methysergide failed to have an effect. In addition to providing support for the severity hypothesis, these results indicate that both opioid and nonopioid forms of conditional analgesia are subserved by critical alpha 2-adrenoceptors at the spinal level.     

Strain differences in reversal of conditional analgesia by opioid antagonists

The ability of both naloxone and naltrexone to block conditional analgesia produced by shock associated contextual stimuli was determined using rats from three suppliers. Naltrexone (7 mg/kg) was equally effective in reversing analgesia on the formalin test in all rats tested. Naloxone (7 mg/kg) significantly affected analgesia in Long-Evans and Holtzman Sprague-Dawley rats but had no effect on Charles River (CD) Sprague-Dawley rats. Charles River rats also differed from the other groups in the amount of time spent freezing to apparatus cues. These results indicate that genetic factors and the choice of antagonists may complicate distinctions between opioid and nonopioid analgesia.     

Modulation of appetitively and aversively motivated behavior by the kappa opioid antagonist MR2266

MR2266 (MR), an opioid antagonist that binds to kappa and mu receptors, was examined for its ability to influence the aversively motivated behaviors conditioned by electric shock and the drinking induced by water deprivation or the availability of a palatable saccharin/glucose solution. The intraperitoneal (ip) and intracerebroventricular (icv) administration routes were contrasted. After both ip and icv administration, MR was able to reverse conditional analgesia as measured by the formalin test. MR enhanced the Pavlovian conditional freezing response when administered icv prior to shock exposure but reduced freezing if given ip prior to shock. A related benzomorphan-derived opioid antagonist, MR1452, also reduced freezing when given ip prior to shock. MR2266 was a potent antidipsogenic agent when administered ip but had no such effect when administered icv. It is concluded that separable opioid systems are involved in the modulation of appetitively and aversively motivated behaviors.     

Naltrexone fails to increase pain affect in response to inflammatory pain in a novel escape/avoidance paradigm

The non-specific opioid antagonist naltrexone has traditionally been used as treatment for opioid overdose, as well as in research settings as an antagonist to examine opioid and non-opioid mediated analgesia. However, the mechanisms by which this drug operates are not well understood, and its exact effects on sensory and affective pain processes remain uncertain. Various studies have demonstrated that naltrexone behaves in a paradoxical manner, leading to analgesia, no discernable changes, or an increase in pain, depending on the circumstances of the study. This imprecise spectrum of effects leads to difficulty in interpreting results in studies where naltrexone was utilized as an antagonist. Therefore, the purpose of this experiment was to further examine whether naltrexone elicits dose-dependent effects in behavioral tests designed to quantify the sensory and affective components of pain. Naltrexone was not expected to have an effect on carrageenan-induced inflammatory pain in sensory pain measures, but a dose-dependent increase was predicted in behavior related to the affective component of pain. Eighty-eight male Sprague–Dawley rats were used to test these hypotheses by measuring Mechanical Paw Withdrawal Thresholds before and after naltrexone injection and by assessing performance in the Place Escape Avoidance Paradigm test, a novel paradigm to test pain affect, in which naltrexone had not been utilized. The results demonstrated that naltrexone failed to increase place/escape avoidance behavior as anticipated, but rather produced a slight, but non-significant, decrease in escape avoidance behavior. Further research is needed to elucidate the differential effects of naltrexone on various aspects of pain-related behavior.     

The benzodiazepine inverse agonist DMCM as an unconditional stimulus for fear-induced analgesia: implications for the role of GABAA receptors in fear-related behavior.

When the benzodiazepine inverse agonist DMCM (6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylic acid methyl ester) occupies the benzodiazepine recognition site on the GABAA receptor complex, the inhibitory action of gamma-aminobutyric acid (GABA) is attenuated. DMCM acted as an unconditional stimulus for one response associated with fear or anxiety, analgesia, as indicated by a dose-dependent (0.25-1.0 mg/kg) suppression of rats' responses to a formalin injection. This was accompanied by other fearlike responses (defecation and urination). The opioid antagonist naltrexone (1.75-14 mg/kg) did not affect these behaviors. Environmental cues associated with DMCM provoked analgesia and defecation in the absence of the drug. The conditional analgesia was reversed by naltrexone (7 mg/kg). DMCM functions as an unconditional fear stimulus by eliciting fear-related behaviors and conditioning those responses to neutral stimuli. The neural circuitry underlying fear conditioning appears to involve tonically inhibitory GABAergic synapses.     

Stress-induced suppression of natural killer cell cytotoxicity in the rat: a naltrexone-insensitive paradigm

The suppression of natural killer (NK) cell cytotoxicity by footshock stress can be attenuated by opioid antagonists, implicating endogenous opioids in its mediation. A stress paradigm that induces NK suppression that is not blocked by the opioid antagonist naltrexone is reported. This stress paradigm is also shown to cause analgesia and elevated plasma corticosterone levels that are not attenuated by naltrexone. In the first experiment, a significant suppression of NK cell cytotoxicity after forced swimming was demonstrated in Fischer 344 rats treated with either saline or naltrexone, compared with nonstressed controls. Significantly higher corticosterone levels were evident in both stressed groups. In the second experiment, the same stress paradigm was shown to cause significant analgesia in the tail-flick test, whereas no differences were seen between groups pretreated with saline and naltrexone. It is concluded that opioids need not always be involved in the suppression of NK cell cytotoxicity by stress.     

Behaviorally functional opioid systems in infant rats: II. Evidence for pharmacological, physiological, and psychological mediation of pain and stress

In order to determine the behavioral characteristics of the neonatal opioid system during distressful situations, a modification of the hot-plate paw-lick test used on adults was developed for infant rats. Ten-day-old pups were analgesic to heat following morphine administration. Pretreatment with an opioid antagonist prevented the analgesia. Morphine analgesia was significantly greater in pups group isolated from the dam. Saline control pups group isolated from the dam exhibited longer latencies than their nest-housed siblings. Individual isolation for 5 min increased paw-withdrawal latency markedly. This was also naltrexone reversible. This analgesia was not seen when pups were tested directly from the nest or when grouped with other pups for the 5 min. It is suggested that the opioid system(s) for stress and pain are functional in Day 10 rats and short-term isolation from the dam is a probable natural stressor that is modulated by endogenous opioid release.     

Naltrexone-sensitive analgesia following exposure of mice to 2450-MHz radiofrequency radiation.

To determine whether exposure to radiofrequency radiation (RFR) would induce sufficient thermal stress to activate endogenous opioid mechanisms, male Swiss Webster mice were exposed to 10, 15, and 20 mW/cm2 RFR in a 2450-MHz waveguide system for 10 min at specific absorption rates (SARs) of 23.7, 34.6, and 45.5 W/kg, respectively, then tested in the abdominal constriction paradigm. Confinement in the RFR exposure chamber alone did not appreciably alter body temperature but did appear to induce a stress-associated analgesia that was not blocked by naltrexone. Exposure of confined mice to RFR raised body temperature and further increased analgesia in an SAR-dependent manner. The high SAR-induced analgesia, but not the hyperthermia, was blocked by naltrexone. These findings suggest that 1) RFR produces SAR-dependent hyperthermia and analgesia, and 2) RFR-induced analgesia is mediated by opioid mechanisms while confinement-induced analgesia involves nonopioid mechanisms.     

A combination of buprenorphine and naltrexone blocks compulsive cocaine intake in rodents without producing dependence

Buprenorphine, a synthetic opioid that acts at both μ and κ opioid receptors, can decrease cocaine use in individuals with opioid addiction. However, the potent agonist action of buprenorphine at μ opioid receptors raises its potential for creating opioid dependence in non-opioid-dependent cocaine abusers. Here, we tested the hypothesis that a combination of buprenorphine and naltrexone (a potent μ opioid antagonist with weaker δ and κ antagonist properties) could block compulsive cocaine self-administration without producing opioid dependence. The effects of buprenorphine and various doses of naltrexone on cocaine self-administration were assessed in rats that self-administered cocaine under conditions of either short access (noncompulsive cocaine seeking) or extended access (compulsive cocaine seeking). Buprenorphine alone reproducibly decreased cocaine self-administration. Although this buprenorphine-alone effect was blocked in a dose-dependent manner by naltrexone in both the short-access and the extended-access groups, the combination of the lowest dose of naltrexone with buprenorphine blocked cocaine self-administration in the extended-access group but not in the short-access group. Rats given this low dose of naltrexone with buprenorphine did not exhibit the physical opioid withdrawal syndrome seen in rats treated with buprenorphine alone, and naltrexone at this dose did not block κ agonist-induced analgesia. The results suggest that the combination of buprenorphine and naltrexone at an appropriate dosage decreases compulsive cocaine self-administration with minimal liability to produce opioid dependence and may be useful as a treatment for cocaine addiction.     

Sex Differences in Salivary Cortisol Levels Following Naltrexone Administration1

Effects of endogenous opioid peptide blockade by naltrexone on salivary Cortisol levels were examined in healthy men (n= 8) and women (n= 6). Participants received naltrexone (100 mg) during one laboratory session and a placebo pill during another session. Drug order was counterbalanced across participants. Saliva samples were collected 24 hr after each pill was administered. Among women, salivary Cortisol levels significantly increased following naltrexone administration compared with a placebo pill. Naltrexone administration did not alter salivary Cortisol levels in men. Results suggest sex differences in neuroendocrine sensitivity to opioid blockade, a finding that may hold significance with regard to the treatment of alcohol addiction with naltrexone.     

Ontogeny of an endogenous, nonopioid and hormonally mediated analgesic system

Rats of different ages (10-day, 28-day, and 3-month-old) were exposed to cold-water stress in order to activate an endogenous analgesic system. The effects of naltrexone (7 mg/kg) and dexamethasone (.4 mg/kg) were also studied to examine the role of the opioid and hormonal systems in cold-water-induced analgesia. Following cold-water exposure, nociception was measured with the tail-flick procedure for 2 hr. Results revealed that cold water produced a significant level of analgesia in the 10-day, 28-day, and 3-month-old age groups with no differences between age groups. In addition, in each age group naltrexone did not block the analgesia while dexamethasone attenuated the analgesia produced by cold water. The effects of naltrexone and dexamethasone confirm that cold-water immersion activates a nonopioid, hormonally mediated analgesic system in each age group. Thus, this experiment found that the endogenous, nonopioid, and hormonally mediated analgesic system activated by cold water is functional early in the development of the rat. The early development of this hormonally mediated analgesic system is in contrast to the slower development of endogenous analgesia systems that are mediated by the central nervous system.     

Mu opioid receptors in the ventrolateral periaqueductal gray mediate stress-induced analgesia but not immobility in rat pups

Rat pups become immobile and analgesic when exposed to an adult male rat. The aim of this study was to determine whether these reactions are under the control of endogenous opioids and to determine the role of the midbrain periaqueductal gray (PAG), which mediates stress-induced immobility and analgesia in adult animals. In Experiment 1, 14-day-old rats were injected systemically with the general opioid receptor antagonist naltrexone (1 mg/kg), which blocked male-induced analgesia to thermal stimulation but did not affect immobility. In Experiment 2, the selective mu opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP; 50 or 100 ng/200 nl) was microinjected into the ventrolateral and lateral PAG. CTOP suppressed male-induced analgesia when injected into the ventrolateral PAG. Male-induced immobility was not affected by CTOP. Male proximity therefore seems to induce analgesia in rat pups by releasing endogenous opioids that bind to mu opioid receptors in the ventrolateral PAG.     

Cold water stress analgesia in rats: differential effects of naltrexone

Cold water swim (CWS, 2 degrees C, 3.5 min) decreases the responsiveness to nociceptive stimuli in rats. The influence of various parameters of the CWS condition on stress-induced analgesia were evaluated by means of naltrexone effects. Naltrexone dose-dependently (but significantly only at high doses--21 mg/kg) partially antagonized 3.5 minute continuous CWS analgesia. Its effect was proportional to the duration of CWS. Naltrexone (14 mg/kg) significantly antagonized intermittent CWS-analgesia (18 10-sec exposures, 3/min) and enhanced the analgesia induced by 60 consecutive exposures (1 sec each, 12/min). These results demonstrate that naltrexone differentially affects CWS-analgesia, depending on specific parametric conditions of the stressor. In addition to activation of a non-specific naltrexone-insensitive analgesia-inducing system (not reduced by the drug in all the conditions studied) there appear to be three naltrexone sensitive systems: (1) a non-opioid analgesia-inducing system which mediates continuous CWS-analgesia; (2) an opioid analgesia-inducing system, involved in intermittent CWS-analgesia; and (3) a naltrexone-sensitive system which opposes the analgesic effect of 60 consecutive exposures. Thus, a highly specific relationship exists between certain parameters of the cold water stressor and the nature of the mechanisms which subserve the induced analgesia.     

Amniotic fluid ingestion enhances opioid-mediated but not nonopioid-mediated analgesia

Ingestion of amniotic fluid or placenta by rats has been shown to enhance several types of opioid-mediated analgesia: that induced by morphine, footshock, vaginal/cervical stimulation, and late pregnancy. This enhancement has also been blocked by administration of opioid antagonists. The present study was designed to examine further the specificity of the enhancement effect for opioid-mediated analgesia by testing for enhancement following administration of aspirin, a nonopioid analgesic. The formalin test was used as the pain threshold assay. Amniotic fluid or beef bouillon was administered by orogastric tube to rats that were treated either with morphine sulfate or saline, or pretreated with naltrexone, then treated with aspirin or vehicle. Both morphine and aspirin treatments produced analgesia. Amniotic fluid significantly enhanced the analgesia produced by morphine, but did not enhance the analgesia produced by aspirin, further suggesting that the enhancing effect of amniotic fluid ingestion is specific for opioid-mediated analgesia, such as that existing at the start of parturition.     

Conditional analgesia, defensive freezing, and benzodiazepines

When rats are placed in a situation that has come to be associated with footshock through the process of Pavlovian conditioning, they react with the species-specific defensive response of freezing and a reduction in sensitivity to painful stimulation. In the present experiments, the effects of three benzodiazepines on both of these responses were examined. Pain sensitivity was measured with the formalin test. Concurrent observations of formalin-induced recuperative behavior and freezing were recorded while the animals were in the presence of shock-associated contextual stimuli. It was found that midazolam (Experiments 1 and 2), chlordiazepoxide (Experiment 3), and diazepam (Experiment 4) were capable of significantly attenuating the conditional analgesia. Midazolam and diazepam also reduced the freezing response. The finding that these anxiolytic agents attenuate both conditional responses suggests that the freezing and analgesia are mediated by a common fearlike process.     

Effect of cold acclimation and repeated swimming on opioid and nonopioid swim stress-induced analgesia in selectively bred mice

Swiss-Webster mice selectively bred for high swim stress-induced analgesia (SSIA) were exposed to continuous ambient cold (5 degrees C) for 6 weeks or to daily 3-min swims for 14 consecutive days either in 20 or 32 degrees C water. Thereafter, mice subjected to the particular procedure were injected intraperitoneally with 10 mg/kg of naltrexone HCl and were tested for modification of the opioid and nonopioid component of SSIA. SSIA was reduced following swims at either water temperature and was antagonized by naltrexone to greater extent than in nonswimming mice. Thus, the nonopioid (i.e. naltrexone-resistant) portion of the overall SSIA was significantly reduced, whereas the opioid (naltrexone-sensitive) portion became relatively augmented. In contrast, SSIA differed neither in magnitude nor in sensitivity to naltrexone between cold-acclimated and unacclimated mice. Swim hypothermia as well as the nonopioid component of SSIA were decreased after repeated swimming at 32 and 20 degrees C, but remained unchanged after cold acclimation. This argues for the essential role of an extrathermal, probably emotional in nature, factor not only in the elicitation of nonopioid SSIA, but also in the modulation of thermoregulatory processes during swimming. We suggest that the emergency component of swim stress, together with initial moderate hypothermic challenge, first produces the opioid form of SSIA, and subsequently, as the swim continues, also affects the thermoregulatory processes maintaining thermal homeostasis. This causes further increase in swim hypothermia and raises its stressing property to induce the nonopioid form of SSIA.     

Assessment of opiate modulation of pain and nociceptive responding in young adults with a parental history of hypertension

This double blind, placebo-controlled study examined the effects of an opiate antagonist, naltrexone, on nociceptive flexion reflex (NFR) thresholds and subjective pain in individuals with and without a parental history of hypertension. Using a repeated measures design, NFR threshold was repeatedly assessed on two testing days after administration of either placebo or naltrexone. Immediately after NFR threshold was determined, participants rated the level of pain experienced during the preceding NFR assessment, and at the end of each session participants’ electrocutaneous pain threshold was assessed. Two primary findings were obtained. First, individuals with a parental history of hypertension exhibited attenuated pain sensitivity. Second, endogenous opioid blockade was associated with increased pain ratings in women but with increased pain threshold in men. In sum, the present study did not support a direct involvement of the endogenous opioid system in the attenuated pain sensitivity observed in individuals at increased risk for hypertension.     

Exposure to novelty induces naltrexone-reversible analgesia in rats

The exposure of rats for 2 min to an open field, to a small box, or to inhibitory avoidance training in the small box was followed by a mild analgesia measured by the tail-flick method. The analgesia was observed as soon as 10 s after the exposure and lasted between 10 and 30 min. It was not observed in animals previously made familiar with the test situation, and it was reversed by the administration of naltrexone (0.1 mg/kg). The data suggest that novelty per se is a sufficient stimulus to activate an opioid-mediated analgesic stimulus.     

Opioid receptors regulate retrieval of infant fear memories: effects of naloxone on infantile amnesia

The authors examined the role of the endogenous opioid system in infantile amnesia for contextual fear conditioning. Rats that were 18 days of age received an aversive footshock in a novel context. Rats displayed conditioned fear when tested 1 min after training but not 24 hr after training. Systemic injection of the opioid receptor antagonist naloxone prior to test, but not immediately after training, alleviated infantile amnesia. Naloxone also alleviated infantile amnesia when injected prior to test 7 days after training. These effects of naloxone were due to actions on central rather than peripheral opioid receptors and were not due to any tendency of the drug to produce fear or freezing. These results show that central opioid receptors regulate retrieval of fear memories in infant rats. ((c) 2006 APA, all rights reserved).     

Acute and chronic fentanyl administration causes hyperalgesia independently of opioid receptor activity in mice

Although μ-receptor opioids are clinically important analgesics, they can also paradoxically cause hyperalgesia independently of opioid receptor activity, presumably via the action of neuroexcitatory glucoronide metabolites. However, it is unknown whether the commonly used μ-receptor opioid analgesic fentanyl, which is not subject to glucuronidation, can also induce hyperalgesia independently of opioid receptor activity. Thus, here we examined whether fentanyl increases nociception on the tail-withdrawal test in CD-1 mice concurrently treated with the opioid receptor antagonist naltrexone or in opioid receptor triple knock-out mice lacking μ, δ, and κ opioid receptors. For both groups, an acute fentanyl bolus dose (0.25 mg/kg, s.c.) and continuous fentanyl infusion (cumulative daily dose: 10 mg/kg) did not cause analgesia at any time. Instead, fentanyl significantly decreased withdrawal latencies relative to pre-drug values for the next 15–60 min and for six days, respectively. MK-801 blocked and reversed hyperalgesia caused by the acute injection and continuous infusion of fentanyl, respectively, in naltrexone-treated CD-1 mice, indicating the contribution of NMDA receptors to fentanyl hyperalgesia. These data show that the synthetic opioid fentanyl causes hyperalgesia independently of prior or concurrent opioid receptor activity or analgesia. Since the biotransformation of fentanyl does not yield any known pronociceptive metabolites, these data challenge assumptions regarding the role of neuroexcitatory metabolites in opioid-induced hyperalgesia.