Hanan Frenk

Pro- and anticonvulsant actions of morphine and the endogenous opioids: Involvement and interactions of multiple opiate and non-opiate systems

The proconvulsant actions of high doses of systemic morphine are probably mediated by 3 different systems. One of them produces non-convulsant electrographic seizures and can be activated separately from the others both by intracerebroventricular injections as well as microinjections into discrete subcortical areas. The enkephalins and beta-endorphin, when administered to the same loci, produce similar effects. Pharmacological evidence suggests that specific opiate receptors of the delta-subtype mediate the epileptiform effects produced by this system. The second system mediating proconvulsant effects of systemic morphine is not mediated by stereo-specific opiate receptors. It produces behavioral convulsions, and the GABA-ergic system has been implicated in its action. A third proconvulsant action of systemic morphine can be activated separately from the other two systems by administering this compound with other convulsive agents or manipulations. Specific mu-type opiate receptors are implicated in this effect. In addition to potent proconvulsant effects, systemic morphine also has anticonvulsant properties which are mediated by specific opiate mu-receptors. The conditions under which morphine acts as a proconvulsant rather than an anticonvulsant agent are, as yet, not understood.

The suppressant effects of naloxone on food and water intake in the rat

Systemic administration of naloxone to food- or water-deprived male, Sprague-Dawley rats suppresses food and water intake in a dose-dependent fashion. Administration of as little as .1 mg/kg naloxone significantly reduces water intake. Morphine also reduces water intake in a dose-related manner. Nonetheless, when morphine and naloxone are given together at various doses, a competitive interaction is seen, suggesting that the suppressant effects of these drugs are, in part at least, mediated by the same opiate receptors. High (10 mg/kg) but not low (.1 mg/kg) doses of naloxone induce conditioned taste aversion; thus, sickness may be the cause of the suppressant effects of high, but not low, doses of the drug. These findings are discussed in relation to the hypothesis that endogenous opioid substances mediate drive-reduction reward.

Continuous co-administration of dextromethorphan or MK-801 with morphine: attenuation of morphine dependence and naloxone-reversible attenuation of morphine tolerance.

&NA; N‐Methyl‐ d‐aspartate (NMDA) receptor antagonists have been repeatedly shown to attenuate the development of opiate tolerance and dependence in rodents. In the present experiments, continuous subcutaneous infusion of either MK‐801 (0.01 mg/kg/h but not 0.005 mg/kg/h) or DM (0.133, 0.67 and 1.33 mg/kg/h) reliably prolonged the antinociceptive effect of continuous subcutaneous infusion of morphine sulfate (2.0 mg/kg/h), indicating attenuation of the development of morphine tolerance. Furthermore, this prolonged antinociception was completely reversible by naloxone (10 mg/kg, i.p.). Doses of MK‐801 and DM that were equipotent in attenuating morphine tolerance (0.01 mg/kg/h and 1.33 mg/kg/h, respectively) revealed different profiles of effects, however, on locomotor activity and naloxone‐precipitated abstinence/withdrawal symptoms. With regard to locomotor activity, rats having received continuous (48 h) subcutaneous infusion of morphine sulfate and MK‐801, but not rats having received morphine sulfate and DM, displayed a reliable and striking increase in locomotor activity as compared with rats having received morphine alone. With regard to naloxone‐precipitated withdrawal symptoms, continuous (48 h) subcutaneous co‐infusion of either MK‐801 (0.01 mg/kg/h) or DM (1.33 mg/kg/h) with morphine attenuated naloxone‐precipitated hyperalgesia as compared with rats infused with morphine alone. MK‐801 (0.01 mg/kg/h) was more effective than DM (0.133, 0.67, or 1.33 mg/kg/h), however, in reducing other naloxone‐precipitated withdrawal symptoms (teeth chattering, jumping and wet dog shakes). The effects of MK‐801 on all withdrawal symptoms were confounded, however, by the appearance of flaccidity following naloxone administration to rats having received MK‐801 and morphine. These results extend previous observations by showing that the prolonged antinociception observed following co‐administration of morphine and an NMDA antagonist is completely naloxone‐reversible, supporting the notion that this antinociception reflects prolongation of an opioid receptor‐mediated effect. The different profiles of side effects associated with MK‐801 and DM, however, suggest that (1) attenuation of naloxone‐precipitated withdrawal symptoms by MK‐801 may be an artifact of toxicity, and (2) DM may prove clinically useful for the prevention of morphine tolerance, given its lack of observable side effects when administered concurrently with morphine to rodents.

Cholecystokinin antagonists selectively potentiate analgesia induced by endogenous opiates

We have recently observed that exogenous sulfated cholecystokinin octapeptide (CCK) can antagonize various forms of opiate analgesia and that the CCK receptor blocker proglumide potentiates morphine analgesia. These observations, plus the similarity in the distribution of CCK and opiate systems, suggest that endogenous CCK may act as a physiological opiate antagonist. We have extended these initial studies by examining the effect of CCK antagonists on opiate analgesia produced by release of endogenous opiates (front paw footshock induced analgesia) and by intrathecal administration of D-Ala-methionine enkephalinamide, a stable analogue of an endogenous opiate. Additionally, the specificity of proglumides effect was examined by testing the effect of this drug on various forms of non-opiate analgesia. This series of experiments demonstrate that CCK antagonists can markedly potentiate analgesia induced by endogenous opiates and provide strong support for the hypothesis that endogenous CCK systems can oppose the analgesic effects of opiates. Potentiation of analgesia by CCK receptor blockers appears to be selective for opiate systems since proglumide typically attenuated or had no effect on various forms of non-opiate analgesia. These data suggest that CCK blockers may be clinically useful for enhancing the analgesic effects of procedures such as acupuncture, which may be mediated by release of endogenous opiates.

Is substance P a primary afferent neurotransmitter for nociceptive input? I. Analysis of pain-related behaviors resulting from intrathecal administration of substance P and 6 excitatory compounds

Intrathecal (i.t.) injection of substance P (SP), capsaicin, kainic acid, picrotoxin, strychnine, morphine, and L-glutamic acid in rats induced rhythmic scratching movements with the hindlimbs, biting, and, with some of these compounds, vocalization and myoclonic twitches. Although biting was directed to the dermatome corresponding to the injection site, scratching was aimed at anterior dermatomes. Presumably painful chemical stimulation produced by cutaneous and subcutaneous application of capsaicin or acetic acid never elicited scratching. Vocalization was never elicited by SP. When vocalization occurred following i.t. picrotoxin and morphine, it was correlated with myoclonic twitches rather than with scratching and/or biting. These findings indicate that scratching (a) is not pain-related and, (b) when elicited by the i.t. administration of the compounds listed above, does not result from activation of nociceptive primary afferent synapses.

Development of thermal nociception in rats

&NA; In the present study, we show that hypersensitivity to noxious thermal stimulation can be seen clearly in developing rats. Rats, at postnatal days 3, 6, 9, 12, 15, 21 and 90 were tested for reflex responsiveness to noxious heat, using tail withdrawal from hot water as the assay. Thermal nociceptive thresholds are considerably lowered, relative to adults, up to postnatal day 12. Thresholds were 39, 37.5, 40.8, 43.3, 46.5, 45.2 and 47.2°C for the respective age groups. Enhanced sensitivity to suprathreshold noxious stimuli is seen in neonates up to postnatal day 15 (but not on day 9). Starting on day 21, sensitivity to noxious stimuli decreases with increasing age, as can be seen by the decrease in the slope of the temperature‐response curve (system gain). Spinal transections at postnatal days 13, 17, 20, 60, or 100 did not produce a change in nociceptive thresholds in any of the age groups. In contrast, sensitivity to noxious stimulation (system gain) was enhanced by spinalization in rats 20 days of age or older. Based on these results we suggest that threshold elevation with increasing age most probably reflects changes in local spinal properties, while changes in responsiveness to suprathreshold noxious stimuli involves maturation of both spinal and descending supraspinal structures.

Effects of the combined oral administration of NSAIDs and dextromethorphan on behavioral symptoms indicative of arthritic pain in rats.

&NA; The effects of combined single oral treatments with non‐steroidal anti‐inflammatory drugs (NSAIDs) and the non‐competitive N‐methyl‐ d‐aspartate (NMDA) receptor antagonist dextromethorphan (DM) on arthritic pain were examined in a rat model of adjuvant‐induced arthritis. Although 12.5–100 mg/kg doses of DM alone produced no reliable effects, treatments with ibuprofen (IB, 50 and 100 mg/kg but not 12.5 or 25 mg/kg) produced mild analgesia in arthritic rats as determined using the Randall‐Sellito test. IB showed a dose‐response relationship which appeared to plateau at doses of 50 and 100 mg/kg. Adding 50 mg/kg DM to each IB dose resulted in significantly greater analgesic activity than IB alone at doses of 25, 50 and 100 mg/kg. A similar interaction between 50 mg/kg DM and 50 mg/kg IB occurred with respect to spontaneous pain behavior. Adding 25 mg/kg DM to 25 mg/kg IB likewise increased analgesia as measured by both the Randall‐Sellito and spontaneous pain behavior tests (both P < 0.05). Five more NSAIDs were evaluated using the Randall‐Sellito test, which included naproxen (NP), piroxicam (PIR), etodolac (ET), diclofenac (DC), and ketorolac (KE). For all six NSAIDS, the addition of 50 mg/kg DM reliably increased their analgesic potency, as indicated by reliable increases in previously effective NSAID doses (all six NSAIDs) as well as previously ineffective NSAID doses (IB, NP, DC, and PIR). These data demonstrate that DM greatly potentiates the analgesic activity of IB, DC, NP, PIR, ET, and KT and increases the peak effect over the NSAIDs alone. Similar to DMs previously demonstrated enhancement of opioid analgesia in acute pain, the combination of DM and an NSAID may represent a novel analgesic approach to improved management of arthritic pain.

The development of stimulation-produced analgesia (SPA) in the rat

The present study studied the development of stimulation produced analgesia (SPA) from the periaqueductal gray (PAG) in rats. A monopolar stimulating electrode was lowered into the dorsal or ventral PAG of animals aged 7, 14, 21, or 90-120 days. Constant current cathodal pulses (100 Hz, 100 microseconds) were delivered, starting 10 s before analgesia was tested by the tail-flick (TF) test and continuing throughout each TF trial or until cut-off (7 s). Current intensity was increased stepwise (3-200 microA). It was found that SPA can be elicited starting at 21 days, but not earlier. However, supraspinal modulation of nociception is still immature at 3 weeks after birth. First, stimulation intensities needed to produce SPA are higher in 21-day-old pups than in adult animals. Second, in 21-day-old pups, but not in adults effective current intensities in the dorsal PAG are higher than in the ventral PAG. Third, naltrexone decreases SPA from the ventral PAG in 21-day-old pups, but not in adult animals. These findings indicate that supraspinal modulation of nociception develops only 3 weeks after birth, with the ventral PAG maturing prior to the dorsal PAG, and that the contribution of endogenous opioids to SPA does not remain constant throughout the ontogeny of rats.

Different opioid systems may participate in post-electro-convulsive shock (ECS) analgesia and catalepsy

Administration of electroconvulsive shock (ECS) to rats results in post-ictal analgesia and catalepsy both of which can be partially reversed by the opiate antagonists, naloxone and naltrexone. Tolerance to both phenomena develops following daily ECS administration for 10 days. However, qualitatively different patterns of tolerance development of analgesia and catalepsy are seen. Naloxone treatment prior to ECS provides partial protection against the development of tolerance to ECS-induced catalepsy but does not present the tolerance to post-ECS analgesia. In contrast, the long-lasting opiate antagonist, naltrexone, blocked the development of tolerance to ECS analgesia. Furthermore, the same animals that showed tolerance to the analgesic effects of repeated ECS failed to show analgesia following the administration of 10 mg/kg of morphine while naltrexone, but not naloxone, treatment prior to ECS blocked the development of cross-tolerance to morphine analgesia. A dose-response investigation of morphines action (5, 10, 15 and 20 mg/kg) in additional animals receiving 10 daily administrations of ECS reveals that a greater tolerance to morphines motor inhibitory effect than to its analgesic effect results from repeated ECS administration. Finally, animals receiving daily injections of either a low (10 mg/kg) or a high (100 mg/kg) dose of morphine for 10 days showed markedly attenuated post-ECS analgesia and catalepsy. However, whereas similar effects of opiate antagonists and the chronic administration of both doses of morphine were observed with post-ECS catalepsy, analgesia was least affected by naloxone (50% of control) and most affected by the chronic high dose of morphine (14% of control). Furthermore, a similar degree of tolerance to post-ECS analgesia was seen following either repeated ECS in drug-naive animals or the chronic administration of the high dose of morphine. Thus, the partial naloxone blockade of ECS analgesia and the more substantial attenuation of post-ECS analgesia seen in morphine-tolerant animals provide different estimates of opioid involvement in these phenomena. It is proposed that these results may demonstrate the involvement of different opioid systems in analgesia and catalepsy and it is suggested that more than one opioid system may also be involved in post-ECS analgesia.

Calcitonin gene-related peptide enhances substance P-induced behaviors via metabolic inhibition: in vivo evidence for a new mechanism of neuromodulation

The present study examined the effects of intrathecal (i.t.) injection of calcitonin gene-related peptide (CGRP) on caudally directed biting and scratching induced by i.t. substance P (SP), bombesin (BBS), strychnine (STR), and kainic acid (KA). CGRP alone (5.25, 10.5 and 21 nmol) had no effect on these behaviors, but CGRP pretreatment produced a dose-related enhancement of behaviors induced by SP or BBS, but not by KA or STR. 2-Amino-5-phosphonovaleric acid (APV, 25 nmol), a selective N-methyl-D-aspartate (NMDA) receptor antagonist, did not block the CGRP potentiation of SP and BBS induced behaviors. CGRP, however, failed to enhance scratching and biting induced by a SP analogue [pGlu5-Mephe8-MeGly9]SP(5-11) (Dime-C7) that is resistant to enzymatic degradation by SP endopeptidase. These findings demonstrate that CGRP potentiates SP induced behavioral responses via inhibition of neuropeptide degradation and that this mechanism may serve as a physiological mechanism of SP modulation.