Physiological characteristics of pressure immobility. Effects of morphine,naloxone and pain

This study is an attempt to detect the most important modifications of physiological parameters occurring during pressure immobility in rabbits and to compare them with those recorded during animal hypnosis. Like the latter, pressure immobility is characterized by the development of high voltage slow waves in the EEG, reduction in frequency and amount of rhythmic slow activity in the hippocampus (RSA) and depression of spinal polysynaptic reflexes. Systolic and diastolic blood pressures are not modified. Duration of two types of immobility is positively correlated within individuals. Treatment by a single dose of morphine (1 mg/kg) potentiates the duration and this effect is antagonized by naloxone (1 mg/kg). Repeated morphine injection up to tolerance reduces duration. Pressure immobility may also be produced under persistent nociceptive stimulation and is characterized by the development of high voltage slow waves in the EEG, as is typical in the absence of pain. Naloxone, (5 mg/kg) injected in a condition of persistent noxious stimulation, reduces immobility duration. In contrast to animal hypnosis, the duration of pressure immobility is neither potentiated by pain nor reduced by naloxone (1,5 or 20 mg/kg). It is suggested that the two immobilities are controlled by several mechanisms, some similar, some different.     

Topical application of morphine to the rat somatosensory cortex produces analgesia to tonic pain

Topical application of 0.01 or 0.1% morphine solution to the somatosensory SI area of the rat cerebral cortex significantly decreased the pain intensity rating in the formalin test without producing motor side effects or sensory deficits. Naloxone partially antagonizes this effect. Topical application of morphine to the striate cortex did not induce analgesia. It is suggested that the primary somatosensory SI area of the cerebral cortex plays a role in opiate pain control.     

Hippocampal rhythmic slow activity (RSA) during animal hypnosis in the rabbit

Hippocampal electrical activity has been studied in 12 unanaesthetized, unrestrained rabbits during hypnosis and spontaneous activity in the experimental cage, before and among hypnosis trials. Quantitative analyses showed that rhythmic slow activity (RSA) occurred during exploratory movements (mean frequency 7.2 Hz) and also during spontaneous immobility, but at a lower percentage and frequency (6.5 Hz). RSA was always present during the induction of hypnosis, but its frequency decreased and disappeared at the beginning of immobility, when it was replaced by a large amplitude irregular activity (LIA). During hypnosis RSA occurred in short periods, at low frequency (5.7 Hz). During the initial part of hypnosis, the lowest percentage of RSA was recorded. In the second and third part, RSA increased in number of episodes, total amount, duration of a single episode and mean frequency. RSA of higher frequency occurred at the end of hypnosis, preceding righting movements. The relationships between hippocampal RSA and animal hypnosis are discussed.     

Identity of sensory and motor systems that are critical to the immobility reflex (“animal hypnosis”)

This review presents an analysis of the sensory and motor mechanisms as they are now understood that cause the immobility reflex (IR). Of the sensory systems that conceivably could trigger and sustain the IR, as commonly induced experimentally by inversion and manual restraint, evidence has been presented to eliminate some senses (vestibular, vision, sound, many visceral sensations, olfaction, taste, temperature), while incriminating tactile and proprioceptive influences. Of the motor systems which could cause the profound immobility during IR, neurosurgical and electrophysiological evidence identifies the locus of the inhibitory neurons in the brain stem and/or spinal cord. The evidence reviewed leads to a unified working hypothesis of IR mechaisms. IR is considered to be caused by a group of neurons in the brain stem which inhibit spinal motoneurons, either directly or indirectly, when those inhibitory neurons are activated by a specific pattern of tactile and proprioceptive input. Modulation of the IR control system appears to come from the limbic system, which under fear-producing conditions, potentiates the IR in part by release of epinephrine. Inhibition of the IR control system appears to come from the neocortex, as well as the brain stem reticulum, when it is activated by nonspecific, arousing somaesthetic sensations that produce generalized activation of the neocortex and skeletal muscle.     

Depth-duration profile of the immobility reflex: Theoretical implications for its triggering,sustaining, and terminating mechanisms

Immobility reflex (IR) durations of rabbits were remarkably consistent (weighted average range of 0.36–0.70 minutes/trial), under the special conditions when rabbits were “habituated” by approximately 25 preliminary trials and when termination of IR was operationally defined as the point at which any attempt, even abortive, at righting occurred. When the mean duration for each rabbit was normalized (into quartile duration points) and then rabbits were tested at these points to determine arousal threshold, each rabbit revealed a clear progressive decrease in the “depth” of IR as the duration of a given episode progressed. Both duration and depth were decreased when body hair was removed. The hippocampal EEG revealed persistent theta activity throughout a given trial; but the specific frequency the theta increased briefly just prior to induction of IR and decreased immediately afterwards. In Short-duration trials, theta frequency increased progressively until the IR terminated spontaneously. In long-duration trials frequency changes oscillated. Each of these observations in interpreted to support the theory that, during IR, reverberating neural circuits (in the brainstem reticular formation) are activated and their output inhibits spinal motor neurons.     

Effects of morphine and naloxone on the K-stimulated release of methionine-enkephalin from slices of rat corpus striatum

Methionine-enkephalin (ME) released from superfused slices of rat corpus striatum was estimated by radioimmunoassay (RIA). The basal release of2.5 ± 0.2pmol/g/min (0.15% of content per min) was increased approximately 3-fold upon exposure of tissue to 30 mM K⁺ for 5 min. This increase in release was not observed in the absence of Ca²⁺. Both morphine (10⁻⁵ M) and (−)-naloxone (10−⁵ and 10−⁶ M) significantly depressed the release of ME evoked by 30 mM K⁺ but did not alter basal release. The +-isomer of naloxone, which lacks opiate antagonist activity, did not affect basal or evoked release. A consistent depression of release was not observed when 47 mM K⁺ was used to evoke the release of ME. The issue of whether a feedback mechanism controls the release of ME from the striatum cannot be resolved until it is known whether the effect of morphine and naloxone on ME release are mediated by opiate or non-opiate mechanisms.     

Analgesia produced by low doses of the opiate antagonist naloxone in arthritic rats is reduced in morphine-tolerant animals

In a model of experimental chronic pain (adjuvant-induced arthritic rats), low doses of the opiate antagonist naloxone produced a profound analgesia. Maximum analgesia was seen with 3 micrograms/kg (i.v.). In contrast, hyperalgesia was obtained with much higher doses (1-3 mg/kg, i.v.). The hyperalgesic effects were not affected in arthritic animals rendered tolerant to morphine, but the paradoxical analgesic effects were significantly reduced. This decrease suggests that naloxone analgesia involves interaction with opiate receptors and that the operation of endorphinergic systems differs in normal animals and animals which experience persistent pain.     

Alteration of morphine withdrawal to naloxone by interferon

The opiate abstinence syndrome represents a fundamental feature of the addictive process. The present study demonstrated that: 1) recombinant leukocyte A Interferon (alpha-IFN) injection prior to chronic morphine treatment reduces addiction liability; and 2) alpha-IFN and not human gamma-IFN injection to morphine dependent rats significantly modifies the naloxone-induced abstinence syndrome in a characteristic dose response manner. Two hypotheses concerning IFN's action were discussed.     

Effects of morphine and naloxone on cerebral blood flow and metabolism in experimental subarachnoid hemorrhage

Objective - Naloxone is reported to improve the clinical condition of patients with subarachnoid hemorrhage (SAH). If this effect is vascular determined is unknown, wherefore the influence of morphine and naloxone on cerebral blood flow (CBF) and metabolic rate of oxygen (CMRO_z) after SAH was evaluated. Material and methods - Two groups of 8 rats each with SAH and 2 corresponding groups of controls were investigated. CBF was calculated by the intracarotid ¹³³Xenon method and CMRO₂ as the product of CBF and the difference between systemic arterial and cerebral venous oxygen content. Results - In controls morphine, 1 mg/kg administered intravenously, decreased CBF by 25% ( P < 0.001) without changing the CBF/CMRO₂ ratio. In animals with SAH CBF was decreased by 32% ( P < 0.001) and CBF/CMRO₂ ratio by 38% ( P < 0.01). Naloxone, 40ug/kg administered intravenously neither influenced CBF nor the CBF/CMRO₂ ratio in the 2 groups. Conclusion - The reported clinical effect of naloxone after SAH can, according to our results, not be explained by changing the relationship between CBF and metabolism.     

Method of administration determines the effect of naloxone on pain

The opiate antagonist, naloxone, produces dose-dependent biphasic changes in clinical pain. The mechanism of the analgesia produced by low dose naloxone is unknown. To study the analgesic effect of naloxone, we have used a programmable infusion pump, which eliminates placebo-induced endorphin-mediated analgesia, to administer different doses of naloxone. We report that use of machine infusion of naloxone exclusively produces analgesia. The implications of this finding to the mechanism of naloxone-induced analgesia are discussed.     

Increase in the threshold of a nociceptive test induced by naloxone in morphine-tolerant rats

The effects of various doses of naloxone (3–1000 μg/kg i.v.) on the vocalization threshold elicited by pressure on the paw were evaluated in rats chronically treated with high doses of morphine. In addition to the well known precipitation of withdrawal induced by naloxone, an unexpected dose-related increase in the vocalization threshold was observed.     

Single nerve capsaicin: Effects on pain and morphine analgesia in the formalin and foot-flick tests

Application of capsaicin to the sciatic nerve reduces responsiveness to pain in the foot-flick test which examines bried, threshold-level pain. The purpose of the present study was to determine if a similar reduction occurs in the formalin test which examines supra-threshold, deep pain that persists for several hours.The sciatic nerve on one side in the rat was exposed and soaked for 15 min in a solution of capsaicin and the saphenous nerve was ligated and cut. The operated foot was tested for sensitivity to pain in the formalin and foot-flick tests 2 days to 12 weeks later both with and without morphine. The capsaicin treatment produced a substantial reduction in sensitivity to foot-flick heat pain at all times after surgery. In the formalin test, the effects were small and tended to suggest that the rats felt more rather than less pain. The capsaicin treatment markedly reduced the sensitivity of formalin test pain to morphine. This effect appeared about one week after surgery and persisted for 12 weeks. The results suggest that capsaicin-sensitive unmyelinated afferents play a role in the threshold-level, non-damaging heat pain, but are not involved in pain resulting from tissue damage. However, these afferents appear to be important for the spinal action of morphine on this type of pain.     

Studies on pain. Effects of morphine on a spinal nociceptive flexion reflex and related pain sensation in man

The nociceptive flexion reflex and the corresponding subjective pain score elicited by sural nerve stimulation were studied in 6 healthy volunteers. A significant correlation was found between the respective recruitment curves of the reflex and of the pain score as a function of stimulus intensity. Consequently, the reflex (Tr) and the pain (Tp) thresholds were found to be almost identical (mean: 10.6 and 10.3 mA, respectively). Similarly, the threshold of the maximal reflex response (Tmr) was very close to that of intolerable pain (Tip): 37.1 and 38.8 mA, respectively. These four parameters were studied before and after intravenous administration of morphine chlorhydrate (0.05, 0.1, 0.2 and 0.3 mg/kg) and subsequent administration of naloxone hydrochloride (0.02 mg/kg; i.v.). While 0.05 mg/kg morphine remained without any effect, higher doses produced an increase in the four thresholds (Tr, Tp, Tmr, Tip). Furthermore, a very significant linear relationship was found between the importance of the increase and the dose of morphine. Morphine also depressed in a dose-dependent fashion, the nociceptive reflexes elicited by a constant stimulation intensity (1.2-1.3 Tr). All these effects were immediately reversed by subsequent naloxone. During all the pharmacological situations, variations in Tr and Tp as well as in Tmr and Tip were found to be very significantly linearly related, indicating a close relationship between the effects of morphine on the nociceptive reflex and on the related pain sensation. These results suggest that, in our model involving a brief 'epicritic' nociceptive stimulus, the mechanisms of morphine-induced analgesia in man can be explained by a depressive effect on the nociceptive transmission directly at a spinal level.     

Neuroendocrine changes following habituation of animal hypnosis in male rabbits

Neuroendocrine changes have been studied in male rabbits during daily repeated inductions of animal hypnosis. All the animals attained the habituation but after different times, showing a different individual sensitivity to the treatment. Once habituation was reached, further treatment did not elicit any recovery. Corticosterone plasma levels gradually increased during treatment, reached a peak at habituation and did not change afterwards. Because corticosterone was not modified after habituation was established, despite further manipulation of the animals, it can be assumed that the increase of corticosterone is due to hypnosis. ACTH did not follow the same pattern of variation as corticosterone: an increase was found at habituation, but this was not statistically significant. Estradiol decreased and the testosterone/estradiol (T/E) ratio increased on the final day with respect to Day 1, whereas testosterone plasma levels were not modified by the treatment. However, there was a general increase of testosterone metabolites (dihydrotestosterone, estradiol) studied in vitro in the anterior hypothalamus of rabbits sacrificed at the end of the treatment. Thus the different responsiveness associated with repeated induction of animal hypnosis changes basal levels of ACTH, steroid hormones and central testosterone metabolism.     

Behavioural responses to novelty in two inbred mouse strains after intrahippocampal naloxone and morphine

Male C57BL/6 and DBA/2 mice were injected intrahippocampally with either naloxone (0.5 μg) or morphine (1.0 μg), or saline vehicle alone and, after 15 min, some 12 behavioural components carried out in a novel environment were recorded for 20 min. Naloxone reduced exploratory rearing responses, wall-leaning and object-sniffing in strain C57BL/6 and augmented these behaviours in strain DBA/2, while morphine depressed the scores in both. In conjunction with previously obtained evidence that the mouse hippocampus contains a genotype-dependent cholinergic mechanism which regulates responses to novelty, these findings support the hypothesis that hippocampal opioid peptides modulate the cholinergic control of exploration in mice, possibly indirectly through GABAergic pathways. In contrast, locomotor activity, defaecation and tail elevation remained practically unaffected by the two drugs, and grooming showed another kind of genotype-treatment interaction, that is to say, after morphine.     

The effects of naloxone, morphine and methionine enkephalinamide on Ia afferent terminations in the cat spinal cord

Naloxone, morphine, Met5-enkephalinamide (MENKA) and procaine were administered microelectrophoretically near extracellularly stimulated extensor muscle group Ia afferent fibres and terminations in the lumbar spinal cord of cats anaesthetized with pentobarbitone sodium. Observations were made of effects on the electrical threshold, on the depolarizing action of GABA or piperidine-4-sulphonate (P4S), and on bicuculline-sensitive primary afferent depolarization (PAD) generated by tetanic stimulation of flexor muscle low threshold afferents. All 4 agents reversibly elevated the threshold of Ia fibres in the dorsal column and Ia terminations in the ventral horn. The depolarizations of terminations by GABA or P4S were also reduced, an effect, which for all except MENKA, probably accounted for a concomitant reduction in PAD. In the absence of a consistent effect on either threshold or depolarization by GABAmimetics, MENKA reversibly diminished PAD, an action blocked by naloxone. Intravenously administered naloxone, in doses known to enhance spinal monosynaptic excitation in the cat, had no effect on GABAergic PAD and little or no effect on Ia termination threshold. The results are discussed in relation to a naloxone-sensitive effect of MENKA which reduces transmitter release from GABAergic axo-axonic synapses on Ia terminals, but which does not account for the enhancement of spinal reflexes by naloxone.     

Spinal pain suppression mechanisms may differ for phasic and tonic pain

The dorsalateral funiculus (DLF) spinal pathway has previously been identified as a major pathway involved in descending modulation of pain. While bilateral lesions of the DLF attenuated systemic morphine analgesia as measured by the tail-flick test, they failed to attenuate analgesia as measured by the formalin test. These results suggest that phasic and tonic pain, modeled by the tail-flick and formalin tests, respectively, may utilize different pain suppression mechanisms.     

Evaluation of the analgesic effects of capsaicin using a new rat model for tonic pain

An animal model for tonic pain has been produced by means of injecting monosodium urate crystals into a knee joint of rat hind paws in order to evaluate the analgesic effects of various drugs and analgesic methods and to elucidate the physiology of tonic pain. This model allows for stable and long-term behavioural changes due to tonic pain followed by complete recovery without tissue damages. It is advantageous in allowing for objective and quantitative evaluation of the effects of analgesics and should prove useful in research on pain and the development of pain therapy techniques. Using this model, the analgesic effects of capsaicin was evaluated when administered to the neonatal rat or locally to peripheral nerves. In both cases, significant analgesic effects were obtained.     

Dose effects of morphine on the spontaneous unit activity recorded from the thalamus,hypothalamus, septum,hippocampus, reticular formation,central gray,and caudate nucleus

Spontaneous activity was recorded from 652 units in 8 subcortical structures of unanesthetized rats. Recordings were obtained in central gray, mesencephalic reticular formation, parafasciculus thalami, caudate nucleus, anterior and ventro- medial hypothalamus, lateral septum, and dorsal hippocampus. Eighty recordings were obtained from untreated animals and 80 from saline-injected controls, none of which showed any significant changes of unit activity during the 4- 5-hr observation period. The effect of morphine, given in 5 incremental doses from 0.5 to 30.0 mg/kg ip, was followed in 492 units. Morphine enhanced or depressed spontaneous discharge rates, or caused biphasic effects, ie enhancement alternating with depression and vice versa. Naloxone induced increase in firing after either effect of morphine, or reduced spontaneous activity after morphineinduced increases. However, when morphine reduced neuronal discharges, naloxone never caused further depression. In 86 units, morphine at any dosage failed to alter neuronal activity, but in 54 of these units naloxone nevertheless induced alterations in firing rates. The pattern of responses to morphine differed between all 8 brain regions examined and was characteristic for each individual structure. This is the first systematic study describing the dose-response characteristics of morphine in 8 brain sites recorded simultaneously. Furthermore, it utilized freely behaving animals without the interference of anesthetics, which are themselves known to interact with opiates. The variety of response patterns seen supports the neuro pharmacological evidence for multiple opiate receptors or multiple sites of opiate action.     

Influence of morphine and cyclazocine on the cortical epileptic foci in rabbits

The effects of morphine, cyclazocine and naloxone on penicillin- and strychnine-induced epileptic foci were studied in rabbits. The intracortical injection of penicillin (75, 150 and 300 units) elicited isolated spikes followed by repeated ictal events. The application of strychnine (0.062 and 0.125%) over the cortical surface of one side induced appearance of ipsilateral spiking spreading to the contralateral cortex.

Administration of morphine (0.25–0.75 mg/kg i.v.) or cyclazocine (0.05–3.0 mg/kg i.v.) inhibited the occurrence or the duration of the EEG and motor manifestations induced by penicillin (75 and 150 units) and strychnine (0.062 and 0.125%), while it did not influence the effect of 300 units of penicillin. High doses of morphine (up to 10 mg/kg i.v.) failed to affect the epileptic responses to penicillin and strychnine and at the same time significantly reduced the pO₂ in arterial blood.

Naloxone per se potentiated the effects of the lower doses of penicillin and strychnine. Only at very high doses (20 mg/kg i.v.) displayed a weak antagonism towards the anticonvulsant effect of the two opiates. A full antagonism is only observed towards the effect of cyclazocine (2 mg/kg i.v.) administered after penicillin.

Present data provide additional evidence of the heterogeneity of regulations by opioids of convulsive phenomena. One can hypothesize that the anticonvulsant effect of the two opiate agonists is mediated by naloxone-insensitive opiate receptors, while the proconvulsant-convulsant effect of naloxone might be related to an inhibition of GABA and glycine-mediated transmission.