Swim Stress-induced Analgesia Research Articles

Mu and delta opioid receptor analgesia, binding density, and mRNA levels in mice selectively bred for high and low analgesia

The present study examined mu and delta opioid analgesia, receptor binding, and receptor mRNA levels in lines of mice from two selective breeding projects of relevance to opioid analgesia. Large differences were observed in the analgesic potency of [ d-Ala 2, NMPhe 4, Gly-ol]enkephalin (DAMGO), [ d-Pen 2,5]enkephalin (DPDPE), and [ d-Ala 2]deltorphin II (DELT), selective mu, delta 1, and delta 2 opioid receptor agonists, respectively, in mice selectively bred for high (HA) and low (LA) swim stress-induced analgesia (SIA). HAR and LAR mice, selectively bred for high and low levorphanol analgesia, respectively, display equally large differences in their analgesic sensitivity to DAMGO, modest differences in sensitivity to DPDPE, and no differences in sensitivity to DELT. These sizable genotypic differences in analgesic potency were accompanied by HA/LA and HAR/LAR differences in whole-brain homogenate [ 3 H ]DPDPE and/or [ 3 H ]DELT, but paradoxically not [ 3 H ]DAMGO, binding. Solution hybridization of mRNA extracts encoding mu (MOR-1) or delta (DOR-1) opioid receptors indicated some regional differences in gene expression between high and low lines. Surprisingly, differences in these in vitro markers were often in the direction of LAR>HAR. The present data indicate that selection for either SSIA or levorphanol analgesia produces differential effects on mu and delta opioid analgesia that are accompanied by alterations on in vitro assays, the significance of which remains to be determined. The data are discussed with regard to the utility of in vitro biological markers and genetic models of analgesia.

Acute morphine dependence in mice selectively-bred for high and low analgesia

Acute morphine dependence was compared in mice selectively-bred for high (HA) and low (LA) swim stress-induced analgesia and high (HAR) and low (LAR) levorphanol analgesia by counting the number of naloxone-precipitated jumps. Whereas LAR mice displayed greater acute morphine dependence than HAR mice, HA and LA mice did not differ. No genotypic differences were observed in non-dependent mice, discounting possible differences in basal naloxone sensitivity and/or opioid peptide levels. Thus, the two selection projects, while both producing lines exhibiting highly divergent sensitivity to morphine analgesia, have not had analogous effects on all opioid measures, supporting the notion of independent genetic mediation of opioid analgesia and dependence. Further, these data suggest that analgesic sensitivity may not predict sensitivity to morphine dependence.

Reduction of stress-induced analgesia following ethanol exposure in mice

In the present study, we examined the effects of ethanol treatment on the subsequent expression of opioid and nonopioid forms of swim stress-induced analgesia (SSIA). In Experiment 1, mice were injected with ethanol (2.5 g/kg, IP) or an equal volume of saline once a day for two days. Animals received no treatment on day 3. On day 4, the animals were tested for opioid (3-min swim in water maintained at 32 °C) or nonopioid (3-min swim in water maintained at 20 °C) SSIA in the hotplate test (52 °C). Mice pretreated with ethanol injections showed a decrease in nonopioid SSIA, but not in opioid SSIA. In Experiment 2, mice were given an ethanol solution (10%) or tap water to drink for 15 days. On day 16, all animals were given tap water to drink. On day 17, the animals were tested for opioid or nonopioid SSIA. Neither form of SSIA was modified in mice that drank the ethanol solution. These results show that ethanol pretreatment can modify nonopioid endogenous analgesic responses in mice. Further, the route of administration influences the effects of ethanol pretreatment on SSIA.

Alterations in swim stress-induced analgesia and hypothermia following serotonergic or NMDA antagonists in the rostral ventromedial medulla of rats

HOPKINS, E., M. SPINELLA, Z. W. PAVLOVIC AND R. J. BODNAR. Alterations in swim stress-induced analgesia and hypothermia following serotonergic or NMDA antagonists in the rostral ventromedial medulla of rats. PHYSIOL. BEHAV. 64(3) 219–225, 1998.—Serotonergic, NMDA, or opioid antagonists in the rostral ventromedial medulla (RVM) reduce morphine analgesia elicited from the periaqueductal gray (PAG). Continuous (CCWS) and intermittent (ICWS) cold-water swims elicit respective naltrexone-insensitive and naltrexone-sensitive analgesic responses. CCWS analgesia is reduced by systemic NMDA receptor antagonism and by systemic, but not intrathecal serotonergic antagonism. ICWS analgesia is reduced by both systemic and intrathecal serotonergic antagonism, but unaffected by systemic NMDA antagonism. The present study evaluated whether serotonergic (methysergide: 5–10 μg) or competitive [AP7 (2-amino-7-phosphonoheptanoic acid): 0.01–0.1 μg] or non-competitive [MK-801 (dizocilipine maleate): 0.3–3 μg] NMDA antagonists in the RVM altered CCWS and ICWS analgesia and hypothermia as well as basal nociceptive latencies. Methysergide in the RVM significantly potentiated CCWS, but not ICWS analgesia. In contrast, AP7 in the RVM significantly potentiated ICWS analgesia. Antagonist-induced changes in either hypothermia or basal nociception failed to account for any alterations in stress-induced analgesia. These data suggest that serotonergic, but not NMDA, receptors in the RVM may mediate collateral inhibition between mesencephalic morphine analgesia and naltrexone-insensitive CCWS analgesia.

Evidence that Adaptation to Cold Water Swim-Induced Analgesia Is a Learned Response

Blustein, J. E., L. Ciccolone and P. J. Bersh: Evidence that adaptation to cold water swim-induced analgesia is a learned response PHYSIOL BEHAV 63(1) 147–150, 1998.—Controlling for novelty of the test context, the present experiment determined if adaptation to forced cold water swim stress-induced analgesia is learned through Pavlovian conditioning. Following baseline measurement of pain sensitivity, Group A swam in Context A for 3 min and, 8 h later, sat in Context B for 3 min. The conditions were reversed for Group B. All rats were given a tail-withdrawal test immediately after swimming or sitting in each context. On the first test day, conducted 24 h after the completion of the adaptation phase, all rats swam in Context A for 3 min, and tail-withdrawal latencies were immediately obtained after the swim. On the second test day, 24 h later, all rats swam in Context B, with tail-withdrawal latencies measured immediately thereafter. Both groups showed significantly less analgesia when tested in the adaptation context in which they previously swam than in the other context. These data provide strong evidence that adaptation to stress-induced analgesia is a learned response.

Metabolic correlates of selection for swim stress-induced analgesia in laboratory mice.

The upper limits of metabolic rates and the links between maximal and resting metabolic rates in vertebrates have recently received a lot of attention, mainly due to their possible relationship to the evolution of endothermy. We measured peak metabolic rates during 3 min swimming in 20 degrees C water (Vo2swim), maximal metabolic rate (Vo2max) in -2.5 degrees C Helox, and basal metabolic rate (BMR) in two lines of mice selected for high (HA) and low (LA) swim stress-induced analgesia (SSIA). We found that exercise combined with heat loss used for producing SSIA also acted as a selection agent, resulting in a 15% HA/LA line difference in Vo2swim. Core body temperature of HA mice (characterized by lower Vo2swim) was also on average 3.2 degrees C lower than that of LA mice. Furthermore, Vo2max of HA mice was lower than that of LA mice by 8% and accompanied by larger hypothermia. Thus mice with exceptionally high (or low) Vo2max tended to have exceptionally high (or low) Vo2swim, resulting in a positive correlation between Vo2swim and Vo2max. All these suggest that selection for SSIA produced genetically correlated responses in both Vo2swim and Vo2max. However, we did not observe HA/LA differences in BMR. Hence, changes in resting and maximum metabolic rates are not necessarily correlated. We hypothesize that the lack of such a correlation was partially due to the modulation of metabolic responses by SSIA.

Sex and Genotype Determine the Selective Activation of Neurochemically-Distinct Mechanisms of Swim Stress-Induced Analgesia

A growing literature documents the important influence of organismic factors such as sex and genotype on pain sensitivity and pain modulation. We recently determined that 3-min forced swims in 15°C water produce non-opioid (i.e., naloxone-insensitive) analgesia in outbred Swiss–Webster mice of both sexes; this form of stress-induced analgesia (SIA) is significantly attenuated by the N-methyl- d-aspartate (NMDA) antagonist, dizocilpine (MK-801) in males, but not females. A pilot study designed to confirm the non-opioid and (in male mice) NMDAergic nature of 15°C swim SIA in the C57BL/6J and DBA/2J inbred strains used widely in gene mapping was conducted, using the hot-plate (54°C) assay of nociception. In female mice of both strains, 15°C swim SIA was insensitive to antagonism by either naloxone (10 mg/kg, IP) or dizocilpine (0.1 mg/kg, IP). In male C57BL/6J mice, the observed SIA was naloxone-insensitive, but was attenuated by dizocilpine. This pattern of results is virtually identical to that obtained using Swiss–Webster mice in this and previous studies. However, male DBA/2J mice displayed SIA that was significantly attenuated by naloxone, but insensitive to dizocilpine antagonism. These findings support the hypothesis that genetic factors and sex, in addition to stressor parameters, can determine the selective recruitment of alternative central mechanisms of pain inhibition.

Differential sensitivity of mice bred for stress-induced analgesia to morphine and ACEA-1011 in the formalin test

The antinociceptive effect of morphine, an opioid receptor agonist, and ACEA-1011, a novel NMDA receptor/glycine site antagonist, was examined in the formalin test in mice selectively bred for high (HA) and low (LA) swim stress-induced analgesia (SSIA). A subcutaneous (SC) injection of formalin produced a biphasic nociceptive response in both lines. HA mice spent more time licking the injected paw than the LA mice in both phases of the formalin test. Morphine was equally potent in the early phase in both lines, but it was more potent in HA mice than in LA mice in the tonic late phase of the formalin test. Similarly, ACEA-1011 produced an equally potent antinociceptive effect in the early phase in both lines; however, the compound was more potent in LA mice than in HA mice in the tonic late phase of the formalin test. These data suggest that in HA mice antinociception in the tonic late phase of the formalin test is mediated largely by an opioid-mediated mechanism, whereas in the opioid-deficient LA line at least a nonopioid-mediated mechanism involving the NMDA receptor is also implicated.

Opioid and Nonopioid Swim Stress-Induced Analgesia: A Parametric Analysis in Mice

Environmental stress causes the activation of two types of endogenous pain inhibitory systems in animals: opioid analgesia is antagonized by opiate receptor blockers (e.g., naloxone and naltrexone), whereas analgesia produced by nonopioid systems is insensitive to such antagonism. A large literature documents that the parameters of the laboratory stressor will determine the neurochemical identity of the resultant analgesia. In rats, low severity stressors produce opioid analgesia and higher severity stressors produce nonopioid analgesia. A recent parametric analysis of swim stress-induced analgesia (SSIA) in the female Quackenbush mouse, however, observed the opposite pattern. The present study is a parametric analysis of SSIA using a range of swim temperatures (15–38°C), swim durations (45 s to 7 min), and genetic models [male Swiss-Webster mice, and mice selectively bred from this outbred strain for high (HA), low (LA), or control SSIA]. We find that in nonselected mice low severity swims (i.e., warm temperature, short duration) produce naloxone-sensitive opioid SSIA, whereas high severity swims (i.e., cold temperature, long duration) produce nonopioid SSIA. This pattern is also seen in HA mice displaying very high analgesic magnitudes, but not in LA mice displaying minimal SSIA. In the selectively bred mice, analgesia and hypothermia from forced swimming are positively correlated, but can be dissociated both genetically and neurochemically. Furthermore, swimming in body temperature (38°C) water produces analgesia without concommitant hypothermia, and the increased magnitude of 38°C SSIA displayed by HA mice over control levels is entirely opioid.

Sex differences in the expression and antagonism of swim stress-induced analgesia in deer mice vary with the breeding season

Swim stress-induced analgesia (SSIA) was examined in photoperiodically induced ‘breeding’ (reproductive) and ‘non-breeding’ (non-reproductive) adult male and female deer mice, Peromyscus maniculatus. Nociceptive responses (50°C, hot-plate) of breeding and non-breeding deer mice were determined after either a 1- or 3-min swim in 20°C water. The 1-min swim induced an immediate and relatively short-lasting naloxone (1.0 mg/kg) insensitive ‘non-opioid’-mediated SSIA that was antagonized by the N-methyl- d-aspartate (NMDA) antagonist, MK-801 (0.10 mg/kg) in all of the groups of mice except the breeding (reproductive) females. Breeding females displayed a non-opioid analgesia that was insensitive to MK-801. The 3-min swim induced a relatively more prolonged mixed opioid and ‘non-opioid’ SSIA of which the initial portion was sensitive to antagonism by MK-801 in all groups of the mice except the breeding females, while the latter portion (15 min after swim) was reduced by naloxone in all of the groups of mice. Overall, the breeding males displayed greater levels of SSIA than the breeding females, with no consistent sex differences in the non-breeding mice. Within sexes, the breeding males displayed greater levels of opioid and non-opioid SSIA than the non-breeding males, while the non-breeding females displayed greater levels of SSIA than the breeding females. These results show that both sex and reproductive status affect the expression and neurochemical mediation of non-opioid SSIA. These findings also suggest that reproductive females may have an unique or novel hormonally (estrogen) dependent mechanism associated with the expression of SSIA.

Neonatal testosterone exposure influences neurochemistry of non-opioid swim stress-induced analgesia in adult mice

The effects of neonatal hormone manipulations on swim stress-induced analgesia (SSIA) magnitude and neurochemical quality were examined in Swiss-Webster mice of both sexes. Previous research has indicated that non-opioid SSIA mechanisms in adult Swiss-Webster mice are sexually dimorphic. Male mice exhibit non-opioid SSIA following a 3-min swim in cold (15°C) water that is antagonized by the non-competitive NMDA antagonist MK-801 (dizocilpine; 0.075 mg/kg), whereas female mice do not display NMDA-mediated analgesia in the presence of estrogen. Since male and female mice show equipotent magnitudes of SSIA, it was concluded that female mice display a neurochemically distinct, estrogen-dependent SSIA mechanism specific to their gender. In the present study, female mice exposed to testosterone during the neonatal period display NMDA-mediated analgesia even in the presence of estrogen in adulthood. Thus, expression of the female-specific, estrogen-dependent SSIA mechanism previously described may be dependent on the absence of testosterone during early ontogeny.

Oligogenic determination of morphine analgesic magnitude: a genetic analysis of selectively bred mouse lines.

Two ongoing selective breeding projects have produced mice that display divergent analgesic responses to morphine. These two projects have selected for similar phenotypes: high and low levorphanol analgesia (HAR/LAR lines; Portland, OR) and high and low swim stress-induced analgesia (HA/LA lines; Jastrzebiec, Poland). Evidence suggests genetic commonalities between mice of the two projects. Using a Mendelian breeding protocol, we have recently found that one or two genetic loci predominantly determine the high morphine analgesia exhibited by HA mice. In the present study we demonstrate that the differential morphine analgesia (5 mg/kg, i.p.) displayed by HAR and LAR mice is similarly oligogenic, predominantly determined by two unlinked loci. A complementation analysis, in which the analgesic responses to morphine of the recessive homozygotes of each project (HAR and HA) were compared to those of their hybrid offspring (HAR x HA), revealed that different genetic loci have been fixed in each project. An intriguing bimodal distribution was observed in the HAR x HA population: Some HAR x HA hybrids displayed greater morphine analgesia than either HAR or HA mice, whereas others displayed minimal analgesia. LAR x LA hybrids displayed less analgesia than either LAR or LA mice. The analgesic responses of HAR x LA and LAR x HA mice were comparable to those of their low-line parents. These findings indicate not only that different loci were responsible for producing high morphine responders in each selection project but that these distinct loci can interact synergistically to produce "superhigh" and "superlow" responders.

One or two genetic loci mediate high opiate analgesia in selectively bred mice

The analgesic responses of humans and laboratory animals are characterized by substantial individual differences. The genetic basis of this variability can be studied experimentally in rodents using a program of selective breeding. One such program selected for high (HA) and low (LA) swim stress-induced analgesia (SSIA) on the hot-plate (56°C) test in Swiss-Webster mice. These lines, which have been selectively bred for more than 25 generations, display markedly divergent opioid-mediated SSIA (3-min swims in 38°C water), morphine analgesia (10 mg/kg, i.p.), and analgesia to the κ-receptor agonist, U-50,488H (30 mg/kg, i.p.). The present study investigated the mode of inheritance of these opioid analgesias in HA and LA mice, using Mendelian genetic analyses. We report that the differential sensitivity of HA and LA mice to each of these analgesic manipulations appears to be determined oligogenically, by one or at the most two major genetic loci. The loci associated with each type of analgesia do not co-segregate, however, indicating that three distinct oligogenic effects have been identified. These findings suggest that the genetic determination of analgesic mechanisms may have simple components and as such may be amenable to further analysis using molecular genetic techniques.

Morphine analgesia and tolerance in mice selectively bred for divergent swim stress-induced analgesia

Morphine-induced analgesia and tolerance were examined in Swiss Webster mice selectively bred for high and low swim stress-induced analgesia. Morphine produced a dose-dependent analgesia in both lines; it was 4-fold more potent in the high analgesia line than in the low analgesia line. Despite the differences in morphine-induced analgesia, the degree of tolerance was the same in both lines. Together, these data suggest that selective breeding of mice for high and low swim stress-induced analgesia produced a striking difference in morphine-induced analgesia without affecting the degree of tolerance. Thus, while there is a common genetic determination in swim stress-induced and morphine-induced analgesia, the development of tolerance to morphine possibly relies on a different genetic background.

μ-Opiate receptor binding is up-regulated in mice selectively bred for high stress-induced analgesia

Pain perception and sensitivity to opiate analgesics strongly depend on genotype. Mice selectively bred for high (HA) and low (LA) swim stress-induced analgesia display markedly divergent morphine analgesia, a difference that appears to be determined by one or at the most two major genes. In an attempt to provide candidate genes mediating the supranormal analgesia displayed by HA mice, we performed μ-opiate receptor binding on 27th generation HA, LA, and control (C) mice using [ 3H]naloxone. HA mice were found to have significantly higher whole-brain receptor density ( B max) than LA mice in whole brain homogenates; no significant difference in affinity ( K d) was observed. Quantitative autoradiography confirmed the line difference in whole-brain receptor binding. In the medial thalamus, a brain area implicated in ascending pathways of pain inhibition, HA mice were found to display significantly higher [ 3H]naloxone binding than C mice (a 64% increase) and LA mice (a 128% increase). No significant line differences were observed in any other brain locus. Thalamic μ receptors may therefore play an important role in a central ‘volume control’ mechanism of pain inhibition, and underlie individual differences in the responses of mice to opiate analgesic drugs.

Sex differences in the antagonism of swim stress-induced analgesia: effects of gonadectomy and estrogen replacement

Sex differences in the neurochemical mediation of swim stress-induced analgesia (SSIA) were examined in Swiss-Webster mice. Intact and gonadectomized adult mice of both sexes were tested for their analgesic response (hot-plate test) to 3 min of forced swimming in 15°C and 20°C water. SSIA resulting from 15°C swim was previously shown to be naloxone-insensitive (i.e., non-opioid) whereas SSIA resulting from 20°C swim produced an analgesia that was partially reversible by naloxone (i.e., mixed opioid/non-opioid). The non-opioid components of these SSIA paradigms were attenuated by the N- methyl- d-aspartate (NMDA) receptor antagonist, dizocilpine (MK-801). We now report that in males, but not females, dizocilpine (0.075 mg/kg i.p.) and naloxone (10 mg/kg i.p.) antagonized the non-opioid and opioid components of SSIA, respectively. After ovariectomy, females displayed a pattern of antagonism similar to males such that dizocilpine attenuated non-opioid SSIA, although naloxone remained ineffective in antagonizing 20°C SSIA. Thus, SSIA in intact females was neither opioid- nor NMDA-mediated, yet it was of similar magnitude to the SSIA displayed by intact males. In separate experiments, estrogen replacement (estrogen benzoate; 5.0 μg/day, i.p.) administered to ovariectomized mice over a 6–8 day period reinstated the dizocilpine-insensitivity of 15°C SSIA characteristic of intact females. However, a similar estrogen regimen administered to both intact and castrated males did not compromise the sensitivity to dizocilpine previously noted in male mice. These findings suggest the existence of a novel female-specific estrogen-dependent mechanism of SSIA and highlight the need to consider gender as a factor in basic and clinical research in this area.

Levorphanol and swim stress-induced analgesia in selectively bred mice: evidence for genetic commonalities

Two independent selective breeding programs have developed divergent lines of mice expressing either high and low swim stress-induced analgesia (HA/LA lines; Jastrzebiec, Poland) or high and low levorphanol analgesia (HAR/LAR lines; Portland, OR). In the present study, mice from both programs were tested for both levorphanol analgesia (2 mg/kg) and an opioid-mediated swim stress-induced analgesia (3 min swimming in 32°C water) in the hot-plate test. Mice selected for high and low levorphanol analgesia displayed high and low swim stress-induced analgesia, respectively; mice selected for high and low swim stress-induced analgesia displayed high and low levorphanol analgesia, respectively. This pattern of correlated responses suggests a high degree of common genetic determination in opiate and swim stress-induced analgesia. These findings also suggest that individual differences in analgesic responsiveness to opiate drugs result from genetically determined individual differences in endogenous pain inhibitory mechanisms.

Antagonism of the non-opioid component of ethanol-induced analgesia by the NMDA receptor antagonist MK-801

Recent evidence from our laboratory suggests that the N-methyl- d-aspartate (NMDA) receptor antagonist MK-801 (dizocilpine) selectively antagonizes non-opioid (i.e. naloxone-insensitive) mechanisms of stress-induced analgesia in mice. For example, we have recently demonstrated that a low dose of MK-801 (0.075 mg/kg, i.p.) antagonizes the non-opioid component of a mixed opioid/non-opioid swim stress-induced analgesia (SSIA) resulting from forced swimming for 3 min in 20°C water. Since ethanol-induced analgesia (EIA) has been found to be only partially attenuated by naloxone, we hypothesized that MK-801 would similarly block the non-opioid component of EIA. The effects of MK-801 and of the opioid receptor antagonist naloxone (10 mg/kg, i.p.) on analgesia produced by ethanol (2.5 g/kg in 20% vol/vol, i.p.) were studied in control mice and in mice selectively bred for high (HA) or low (LA) SSIA. HA mice showed significantly more, and LA mice significantly less, EIA than controls. Naloxone and MK-801 significantly attenuated EIA in control and HA mice, and in these lines the combined administration of both antagonists blocked EIA completely. In LA mice, which displayed very little EIA, naloxone but not MK-801 reversed EIA completely. These findings provide additional evidence for the role of the NMDA receptor in non-opioid mechanisms of analgesia. The finding that mice selectively bred for high and low SSIA also display high and low EIA suggests common mediation of the effects of stress and ethanol on antinociceptive processes.

NMDA receptor antagonist MK-801 blocks non-opioid stress-induced analgesia in the formalin test

The analgesic effect of a 3-min swim stress was assessed using the formalin test. Male Swiss mice were injected i.p. with naloxone (0.1 or 1.0 mg/kg), MK-801 (0.075 mg/kg) or saline 15 min prior to swimming in water maintained at 20°C or 32°C. The mice were then injected with 20 μ1 of 5% formalin into the plantar surface of 1 hind paw and pain behaviour (time spent licking the injected paw) was continuously monitored during the subsequent 10 min. Swim stress produced a significant reduction in pain behaviour at both 20°C and 32°C. MK-801 completely blocked the analgesia produced by both the 20°C and 32°C swim. At a dose of 0.1 mg/kg, naloxone partially antagonized the analgesia produced by the 32°C swim but did not affect the analgesia produced by the 20°C swim. Naloxone at a dose of 1.0 mg/kg had no effect on swim stress-induced analgesia. Neither MK-801 nor 0.1 mg/kg naloxone altered baseline pain behaviour, although 1.0 mg/kg naloxone did significantly reduce it. It is unlikely that the effect of MK-801 on swim stress-induced analgesia is due to an interaction with an opioid mechanism, as MK-801 had no effect on morphine analgesia. These results suggest that the analgesia produced by the 20°C swim stress in the formalin test is non-opioid in nature and mediated via the NMDA receptor, whereas the 32°C swim stress-induced analgesia has both an opioid and non-opioid component.

N-methyl- d-aspartic acid (NMDA) receptor antagonist MK-801 blocks non-opioid stress-induced analgesia. II. Comparison across three swim-stress paradigms in selectively bred mice

The effects of the specific N-methyl- d-aspartic acid (NMDA) receptor antagonist MK-801 (dizocilpine, 0.075 mg/kg, i.p.) on swim-stress-induced analgesia (SSIA) were studied in control (C) mice and in mice selectively bred for high (HA) or low (LA) SSIA. In three consecutive experiments, animals were subjected to forced swimming at water temperatures of 20°C, 32°C and 15°C and the resulting analgesia (hot-plate test) was found to be mixed opioid/non-opioid, opioid and non-opioid, respectively, as a function of the degree of antagonism by naloxone (10 mg/kg, i.p.). The major finding of this study is that MK-801 attenuated 15°C SSIA, against which naloxone was ineffective, but had no effect on 32°C SSIA, which naloxone blocked completely. A combination of naloxone and MK-801 significantly attenuated 20°C SSIA in C and HA mice and in HA mice this attenuation was significantly larger than that produced by either drug alone. Morphine analgesia (10 mg/kg, i.p.) was unaffected by MK-801. It is concluded that low doses of MK-801 selectively block non-opioid mechanisms of SSIA.