enkephalin--leucine-2-alanine and Pain

Topics: Animals; Anticonvulsants; Autonomic Nervous System; Blood Pressure; Brain; Diprenorphine; Drug Tolerance; Electroconvulsive Therapy; Electroencephalography; Endorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Heart Rate; Morphine; Naloxone; Pain; Pituitary Gland; Pressoreceptors; Rats; Receptors, Opioid; Sensory Thresholds; Tritium

D-Ala-D-Leu-enkephalin (DADL) is a pentapeptide which, compared to morphine, preferentially binds to the delta receptor. We compared the analgesic and side effects of intrathecal (i.t.) DADL and i.t. morphine sulfate (MS) in 10 tolerant cancer patients with chronic pain at or below the T12 level who were receiving inadequate relief or unacceptable side effects from systemic opiates. These patients were given i.t. DADL and i.t. MS in a randomized, double-blind, cross-over study on separate days at least 1 day apart. I.t. DADL produced analgesia in all patients tested. Total pain relief was greater with DADL than MS in 6 patients, equal in 1 patient and less with DADL in 3. Side effects, most commonly drowsiness, were similar with both MS and DADL and suggest supraspinal effects by both drugs. At the doses given i.t. DADL produced effective pain relief in patients tolerant to systemic opiates although no significant difference in analgesic efficacy between MS and DADL was observed. Studies of the relative analgesic potency of i.t. DADL in man are necessary to fully assess its value in those patients tolerant to systemic or i.t. opiates.

Topics: Adult; Aged; Analgesia; Chronic Disease; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Female; Humans; Male; Middle Aged; Morphine; Pain; Sleep Stages

The negative emotions caused by persistent pain, called affective pain, are known to seriously affect human physical and mental health. The anterior cingulate cortex (ACC), especially the rostral ACC (rACC) plays a key role in the development of this affective pain. N-methyl-d-aspartate (NMDA) receptors, which are widely distributed in the ACC, are involved in the regulation of emotional behavior. It is well known that activation of opioid receptors can relieve pain, but whether it can alleviate affective pain is not clear. In the present study, conditioned place avoidance (CPA) responses induced by complete Freund's adjuvant (CFA) were used to represent the affective pain of place aversion. The behavioral measurements were synchronously combined with multichannel electrophysiological recordings of the discharge frequency of rACC pyramidal neurons to explore whether affective pain could be alleviated by the synthetic opioid [D-Ala2, D-Leu5]-Enkefalin (DADLE), an agonist of δ-opioid receptors. To further investigate this treatment as a mechanism for the relief of affective pain in CFA-treated animals, we used whole-cell patch recordings in slice preparations of the rACC region to determine the dose-dependent effects of DADLE on NMDA receptor-mediated currents. Then, western blot was used to determine levels of phosphorylated NMDA receptor subunits GluN1, GluN2 and GluN3 as affected by the δ-opioid receptor activation. The results showed that activation of δ-opioid receptors down-regulates the phosphorylation of NMDA receptor subunits, thereby inhibiting NMDA currents, decreasing the discharge frequency of rACC pyramidal neurons, and reversing the CPA response. Thus, δ-opioid receptor activation in the rACC region can alleviate affective pain.

Topics: Animals; Enkephalin, Leucine-2-Alanine; Freund's Adjuvant; Gyrus Cinguli; N-Methylaspartate; Pain; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Receptors, Opioid, delta

Topics: Animals; Colon; Enkephalin, Leucine-2-Alanine; HEK293 Cells; Humans; Inflammation; Mice; Nanoparticles; Neurons; Nociceptors; Pain; Receptors, Opioid, delta; Signal Transduction

This study assesses the effects of peripheral or intrathecal pre-treatment or post-treatment with micro, delta, kappa and nociceptin/orphanin FQ (NOP) opioid receptor agonists (morphine, U-50488 [trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl]benzeneacetamide hydrochloride], DADLE [D-Ala2-Leu5-enkephalin] and nociceptin, respectively) on formalin-induced secondary mechanical allodynia and hyperalgesia in rats. 1% Formalin injection produced acute nociceptive behaviors (flinching and licking/lifting) followed by long-term tactile secondary allodynia and hyperalgesia. Neither peripheral (into the formalin-injected paw) nor intrathecal morphine post-treatment reversed formalin-induced secondary allodynia and hyperalgesia. In contrast, morphine pre-treatment prevented the development of these pain behaviors. Intrathecal and peripheral post- but not pre-treatment with U-50488 or DADLE significantly reduced secondary allodynia and hyperalgesia. Interestingly, nociceptin reduced both pain behaviors regardless of the administration site or treatment time. Local antinociceptive effects of morphine, DADLE, U-50488 or nociceptin were blocked by naltrexone, naltrindole, 5-guanidinonaltrindole and [Nphe(1)]nociceptin(1-13)NH(2), respectively. These results suggest that the long-term nociceptive behaviors induced by formalin are differentially modulated by selective opioid receptor agonists. In addition, data suggest that peripheral and spinal delta and kappa opioid receptors are important when nociceptive behaviors are established. In contrast, micro opioid receptors are more important at the beginning of the injury when the sensory system has not changed. NOP receptors participate diminishing both the development and maintenance of nociceptive behaviors. Results suggest that a barrage of afferent input induced by formalin injection initiates a long-term differential change in peripheral and spinal processing that affect the efficacy of opioid receptor agonists.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Analgesics; Animals; Behavior, Animal; Enkephalin, Leucine-2-Alanine; Female; Formaldehyde; Hyperalgesia; Morphine; Nociceptin Receptor; Pain; Rats; Rats, Wistar; Receptors, Opioid

The present study was designed to investigate the roles of different subtypes of opioid receptors in ventrolateral orbital cortex (VLO) opioid-evoked antinociception in formalin test by using an automatic detection system for recording the nociceptive behavior (agitation) and a manual method for detecting the duration of licking the injected paw in the conscious rat. Formalin (5%, 50 microl) s.c. injected into the hindpaw produced a biphasic agitation response or lengthening duration of licking. Morphine (5 microg) microinjected unilaterally into VLO significantly inhibited the agitation response and the licking time, and these effects were blocked by pre-administration of the non-selective opioid receptor antagonist naloxone (1.0 microg) into the same site. Microinjection of endomorphin-1 (5 microg), a selective micro-receptor agonist, and [D-Ala2, D-Leu5]-enkephalin (DADLE, 10 microg), a delta-/micro-receptor agonist also inhibited the nociceptive behaviors, and both the effects were blocked by selective mu-receptor antagonist beta-funaltrexamine hydrochloride (beta-FNA; 3.75 microg), but the DADLE-evoked inhibition was not influenced by the selective delta-receptor antagonist naltrindole (5 microg). Microinjection of selective kappa-receptor agonist (+/-)-trans-U-50488 methanesulfonate salt (1.5 microg) failed to alter the nociceptive behaviors induced by formalin injection. The beta-FNA and naloxone applied into VLO and morphine into the adjacent regions ventral and dorsal to VLO had no effect on the formalin-evoked nociceptive behaviors. These results suggest that mu- but not delta- or kappa-opioid receptor is involved in the VLO opioid-evoked antinociception in formalin test rat.

Topics: Animals; Enkephalin, Leucine-2-Alanine; Injections, Intraventricular; Male; Narcotic Antagonists; Narcotics; Nociceptors; Pain; Pain Measurement; Prefrontal Cortex; Rats; Rats, Sprague-Dawley; Receptors, Opioid

In the light of functional studies, it has been suggested that antibodies directed to alpha subunits of G-proteins delivered into cerebrospinal fluid (CSF) reached and blocked the function of neural transducer proteins. Current understanding indicates that IgGs do not move freely across plasma membranes. Therefore, to characterize the uptake of these antibodies by neural cells, anti-Gi2alpha IgGs were labeled with 125I, fluorescein or with gold particles. The expression of Galpha subunits was also reduced by blocking their mRNA with antisense oligodeoxynucleotides (ODN). Following intracerebroventricular (icv) injection of gold-conjugated anti-Gi2alpha IgGs, electrondense particles entered and became distributed in the cytoplasm and plasma membranes of neural cells. Scattered particles were also found in dendrites and nuclei. Unlabeled IgGs diminished cerebral signals of fluorescein-labeled anti-Galpha IgGs, indicating that this uptake can be saturated. Cerebral radiostaining promoted by in vivo anti-Gi2alpha 125I-IgGs was almost absent in Gi2alpha knocked-down mice, but not after decreasing the quantity of Gzalpha subunits. The immunosignals of CSF anti-Galpha 125I-IgGs, as well as the impairment of opioid-evoked antinociception, were increased by agonist-induced activation of G protein-coupled receptors. The impairing effect of the antibodies on opioid-evoked antinociception was prevented by agents blocking the cellular uptake of proteins, i.e., cytochalasin B, BSA, DMSO, H7, and by down regulation of protein kinase Cbeta1 (PKCbeta1). In mice treated with an ODN to PKCbeta1 mRNA, 125I-IgGs to Gi2alpha subunits remained bound to periventricular structures and did not spread to deeper areas of the CNS. These results indicate that IgGs delivered into the CSF show a saturable binding to Galpha subunits that translocate to the external side of the neural membrane before being internalized by a PKCbeta1-dependent mechanism.

Topics: Analgesics; Analgesics, Opioid; Animals; Autoantibodies; Biological Transport; Brain Chemistry; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, D-Penicillamine (2,5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Epitopes; GTP-Binding Protein alpha Subunits, Gi-Go; Immunoblotting; Immunoglobulin G; Injections, Intraventricular; Iodine Radioisotopes; Male; Mice; Mice, Inbred Strains; Microscopy, Immunoelectron; Morphine; Neurons; Nociceptors; Oligopeptides; Pain; Protein Kinase C; Signal Transduction

Peptides normally do not cross the blood-brain barrier (BBB). Previously, it has been shown that the hexapeptide enkephalin analogue dalargin with polysorbate-80-coated nanoparticles (DAL/NP) can be transported across the BBB and is able to exhibit an antinociceptive effect in mice. In the present study, the circadian time and dose dependencies of the antinociceptive effect of different dalargin preparations were investigated. The active preparation (DAL/NP, 5 mg/kg, 10 mg/kg), as well as a dalargin solution in phosphate buffered saline (DAL/SOL, 10 mg/kg) were injected intravenously to groups of 10-12 inbred DBA/2 mice at 12 different circadian times; mice were synchronized to a light-dark (LD) 12:12 regimen. The antinociceptive effect was determined 15 minutes postinjection by the hot-plate test. Experiments with DAL/NP were repeated using the tail-flick test system at two selected times (08:00 and 20:00) to test for dose dependency (2.5, 5, 7.5, 10 mg/kg). Hot-plate latencies were rhythmic under baseline and after DAL/SOL, with acrophases in the dark phase; DAL/SOL did not influence latency time. In contrast, DAL/NP significantly increased reaction time dose dependently; the maximal possible effect was rhythmic with the 10 mg/kg preparation, with a peak effect in the early light phase. Results were confirmed by the tail-flick test. The experiments demonstrate that an enkephalin analogue coated with nanoparticles can easily cross the BBB and is able to display a dose- and time-dependent antinociceptive effect.

Topics: Analgesics; Animals; Blood-Brain Barrier; Circadian Rhythm; Dose-Response Relationship, Drug; Enkephalin, Leucine-2-Alanine; Hot Temperature; Injections, Intravenous; Male; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Microspheres; Pain; Polysorbates; Reaction Time

The changes in the latency for tail withdrawal in response to noxious heating of the skin induced by microinjection of opioid or serotonergic agonists into the anterior pretectal nucleus (APtN) was studied in rats. The mu-opioid agonist DAMGO (78 and 156 picomol), but not the delta-opioid agonist DADLE (70 and 140 pmol), the kappa-opioid agonist bremazocine (0.24 and 0.48 nanomol) or the sigma-opioid agonist N-allylnormetazocine (0.54 nanomol), produced a dose-dependent antinociceptive effect. The 5-HT1 agonist 5-carboxamidotryptamine (19 and 38 nanomol) and the 5-HT1B agonist, CGS 12066B (1.12 and 2.24 nanomol), but not the non-selective 5-HT agonist m-CPP (41 to 164 nanomol), 5-HT2 agonist alpha-methylserotonin (36 and 72 nanomol) and 5-HT3 agonist 2-methylserotonin (36 and 72 nanomol), produced a dose-dependent antinociceptive effect. These results indicate that the antinociceptive effects of opioid or serotonergic agonists microinjected into the APtN depend on drug interaction with local mu or 5-HT1B receptors, respectively.

Topics: Analgesics; Animals; Benzomorphans; Brain; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Microinjections; Pain; Phenazocine; Piperazines; Quinoxalines; Rats; Rats, Wistar; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu; Serotonin; Serotonin Receptor Agonists

Intracerebroventricular (i.c.v.) administration to mice of IgGs raised against alpha subunits of Gi2 or Gx/z transducer proteins lessened the activation of low Km GTPase induced by morphine, DAMGO and DADLE in P2 membranes from mouse periaqueductal grey matter (PAG). In mice injected with anti Gi2 alpha, DADLE, DPDPE and [D-Ala2] Deltorphin II, but not beta-endorphin-(1-31), antagonized the analgesic activity of morphine. Conversely, following anti Gx/z alpha, morphine antagonized the antinociceptive potency of DADLE. It is concluded that opioids display diverse efficacy at mu-Gi2 and mu-Gx/z complexes to produce supraspinal analgesia in mice.

Topics: Amino Acid Sequence; Analgesia; Analgesics, Opioid; Animals; Cell Membrane; Cerebral Ventricles; Endorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, D-Penicillamine (2,5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; GTP Phosphohydrolases; GTP-Binding Proteins; Immunoglobulin G; Injections, Intraventricular; Kinetics; Male; Mice; Molecular Sequence Data; Morphine; Naloxone; Pain; Peptide Fragments; Receptors, Opioid, mu; Signal Transduction

A polyclonal antiserum directed against the first 16 amino acids of the N-terminal sequence of the murine delta opioid receptor was raised in rabbits. The intracerebroventricular (i.c.v.) injection to mice of the anti delta receptor IgGs impaired the antinociception produced by DPDPE, [D-Ala2]-Deltorphin II, DADLE and beta-endorphin-(1-31) when studied 24 h later in the tail-flick test. Antinociception produced by morphine and DAMGO was fully expressed in mice undergoing this treatment. The selective delta antagonist ICI 174864 (0.8 nmols/mouse, i.c.v.) significantly reduced the antinociceptive activity of opioids to the extent observed after giving the antibodies. ICI 174864 did not decrease further the antinociception that remained after the anti delta receptor serum. The specific binding displayed by 3 nM [3H]-DPDPE was reduced in membranes pre-incubated with the antiserum, whereas no change could be detected for 0.6 nM [3H]-DAMGO labelling mu receptors. This experimental approach revealed the delta component of opioid-evoked supraspinal antinociception in mice.

Topics: Amino Acid Sequence; Analysis of Variance; Animals; beta-Endorphin; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, D-Penicillamine (2,5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; GTP-Binding Proteins; Immunoglobulin G; Immunohistochemistry; Male; Mice; Mice, Inbred Strains; Molecular Sequence Data; Morphine; Narcotic Antagonists; Oligopeptides; Pain; Pain Measurement; Protein Binding; Reaction Time; Receptors, Opioid, delta

The purpose of this study was to determine what effects leucine-enkephalin and D-Ala2-D-Leu5-enkephalin have on both the background and naturally evoked activity of thalamic nucleus submedius neurons responsive to mechanical cutaneous stimulation. Thirty-five neurons in the nucleus submedius were fully characterized during single-unit extracellular recordings as nociceptive, low-threshold mechanoreceptive (LTM) or unresponsive. Micropressure was used to apply the opioids. Eighteen neurons were inhibited; 13 of these were nociceptive and one was LTM. Six units were activated; two of these were nociceptive and three were LTM. The remaining 11 units were unaffected. Opioid responses were tested for antagonism by naloxone in 12 neurons; eight of these responses were antagonized by naloxone. Statistical analyses indicated that the effects of enkephalins on nociceptive neurons were selective for neuronal modality. The opioids also altered the response of some nociceptive neurons to receptive field stimulation. The presence of nociceptive neurons in the nucleus submedius that are selectively inhibited by opioids provides additional support for the involvement of submedius neurons in nociception. The results of this study suggest that this involvement is more than merely transmission of nociceptive input, since the opioids may be selectively modulating the type of information that is transmitted to the cortex.

Topics: Animals; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Evoked Potentials; Membrane Potentials; Micromanipulation; Naloxone; Neurons; Pain; Pressure; Rats; Rats, Sprague-Dawley; Stress, Mechanical; Thalamic Nuclei

The antinociception of opiates is mediated through the activation of opioid receptors in several mid brain and brain stem areas. This paper reports that the forebrain area termed area tempestas (AT), first identified as a convulsant trigger area, is also a component of the endogenous pain suppression system. Unilateral AT application of DAMGO, morphine and U-50,488H in rats at doses in the nanogram range produced marked and dose-dependent increases in the latency to respond to nociceptive stimuli. A lower effect is found after application of DPDPE and DADLE. Antinociception is more evident in the hot plate than in the tail flick test. In the former test, the effect was restricted to the paws contralateral to the hemisphere of injection. Unilateral AT application of naltrexone (4 ng) reduced in the contralateral paws the antinociceptive effect that the bilateral AT application of morphine (20 ng/hemisphere) had induced in both body sides. Unilateral application of naltrexone, (20 ng) ICI 154, 129 (20 ng) and Win 44,441-3 (8 ng) antagonized the antinociceptive effect elicited by the systemic injection of morphine (2.5 mg/kg s), DPDPE (20 mg/kg s) and U-50,488H (20 mg/kg s), respectively. In the hot plate test, the antagonism was found in the paws ipsilateral and contralateral to the hemisphere of injection of the antagonists.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Animals; Anticonvulsants; Azocines; Bicuculline; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, D-Penicillamine (2,5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Escape Reaction; Foot; Hot Temperature; Male; Morphine; Naltrexone; Narcotic Antagonists; Olfactory Pathways; Pain; Phenazocine; Pressure; Pyrrolidines; Rats; Rats, Sprague-Dawley; Reaction Time; Receptors, Opioid; Seizures; Tail

Two novel oxime derivatives of naltrexone, 6-[2-phenylethyl]-oximino naltrexone (NPC 831) and 6-[3-phenylpropyl]-oximino naltrexone (NPC 836) were potent agonists at opioid receptors. Both compounds inhibited binding to all three opioid receptor subtypes with nanomolar affinities. In vivo, NPC 831 and NPC 836 were equipotent to morphine and more potent than the kappa-selective agonist U-50,488H to produce analgesia. ED50 values of 4.02 mg/kg for NPC 831 and 2.24 mg/kg for NPC 836 were generated for inhibition of the tail-flick response in the rat, and ED50 values of 0.05 mg/kg for NPC 831 and 0.02 mg/kg for NPC 836 were calculated for inhibition of the writhing response in the mouse. Bombesin-induced scratching was used to evaluate NPC 831 and NPC 836 for kappa-agonist properties, and the A50, defined as the percent antagonism of the bombesin-induced response, was 1.86 mg/kg for NPC 831 and 0.08 mg/kg for NPC 836, compared to an A50 of 1.54 mg/kg for U-50,488H. These data suggest that NPC 831 and NPC 836 possess potent mu- and kappa-agonist properties in vivo, with NPC 836 being approximately twice as potent as NPC 831 to produce analgesia and 20 times as potent as NPC 831 to inhibit the scratching response produced by bombesin.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Acetates; Analgesics; Animals; Binding, Competitive; Bombesin; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Guinea Pigs; Ligands; Male; Mice; Naltrexone; Oximes; Pain; Pyrrolidines; Rats; Rats, Sprague-Dawley; Reaction Time; Receptors, Opioid; Receptors, Opioid, kappa

Recently, subtypes of the opioid delta receptor have been identified. It is not known, however, if a subtype of opioid delta receptor can be associated with the known modulatory action of delta agonists on mu-mediated antinociception. Thus, the present study has used the delta subtype-selective antagonists, [D-Ala2,Leu5,Cys6]enkephalin (DALCE) (delta 1 antagonist) and naltrindole-5'-isothiocyanate (5'-NTII) (delta 2 antagonist) in an effort to determine whether the positive and negative modulation of morphine antinociception produced by opioid delta agonists was the result of activity at specific subtypes of opioid delta receptors. Intracerebroventricular morphine produced a dose-related antinociceptive effect which was not antagonized by coadministration of the delta antagonist, ICI 174,864, or by pretreatment 24 hr before testing with the DALCE or 5'-NTII. Coadministration with morphine of a nonantinociceptive dose of DPDPE or [D-Ala2,Glu4]deltorphin resulted in a leftward displacement of the morphine dose-effect curve (i.e., positive modulation), whereas coadministration of a nonantinociceptive dose of [Met5]enkephalin with morphine resulted in a rightward displacement of the morphine dose-effect curve (i.e., negative modulation). Both the positive and the negative modulatory actions were antagonized when the experiment was conducted in the presence of the delta antagonist, ICI 174,864, or when the mice were pretreated with the delta 2 antagonist, 5'-NTII. In contrast, pretreatment with the delta 1 antagonist, DALCE, failed to affect either the positive or the negative modulatory actions of these delta agonists on morphine antinociception. The data suggest the involvement of an opioid delta 2 receptor in the modulation of morphine antinociception.

Topics: Animals; Drug Interactions; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Injections, Intraventricular; Male; Mice; Mice, Inbred ICR; Morphine; Narcotic Antagonists; Pain; Receptors, Opioid, delta; Receptors, Opioid, mu

1. It has recently been suggested that opioid antagonists may be divided into those possessing negative intrinsic activity (e.g. naloxone) and those with neutral intrinsic activity (e.g. MR2266). 2. MR2266 was chronically administered to rats by subcutaneous infusion at a dose of 0.3 mg kg-1 h-1 for 1 week. 3. This dose reduced ingestive behaviour and blocked the antinociceptive effects of a kappa-agonist, indicating occupation of opioid receptors in vivo. 4. No supersensitivity could be detected to the antinociceptive actions of mu or kappa agonists, either one or two days after cessation of treatment. 5. No up-regulation of mu, delta or kappa binding sites was observed. 6. Since naloxone induces both supersensitivity and receptor up-regulation under equivalent conditions, the results suggest that negative intrinsic activity may be required for these phenomena to occur.

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Analgesics; Animals; Benzomorphans; Binding, Competitive; Drinking; Eating; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Narcotic Antagonists; Pain; Pain Measurement; Pyrrolidines; Rats; Rats, Inbred Strains; Receptors, Opioid; Sensory Thresholds; Up-Regulation

Previously, we demonstrated the enhanced affinity of opioid receptors for naloxone in striatal slices from morphine-dependent mice. In our present study, binding characteristics of the mu opioid receptor agonists, [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAMGO) and dihydromorphine, the delta opioid receptor agonist, [D-Ala2, D-Leu5]enkephalin (DADLE), and the opioid antagonist, naloxone, were examined in striatal slices from morphine-tolerant and -dependent mice. Striatal slices from mice that were implanted with a morphine pellet for 3,7 and 21 days displayed significant decreases in Kd values (5.1, 4.6 and 5.5 nM, respectively) of [3H]DAMGO when compared to those in slices from control animals that were not implanted or implanted with placebo pellets (9.6 and 9.3 nM, respectively). Also, a significant increase in the binding affinity of naloxone, but not that of dihydromorphine, was observed in striatal slices of mice that were implanted with a morphine pellet for 3 days. Significant increases in the Bmax of delta binding sites in striatal slices of mice that were implanted with a morphine pellet for 3, 7 and 21 days (20.7, 18.1 and 17.7 pmol/mg tissue, respectively) were observed when compared to that in slices from control mice that were implanted with placebo pellets (11.4 pmol/mg tissue). The enhancement in the binding affinity of DAMGO and naloxone and the increased density of DADLE binding sites paralleled the development of morphine tolerance and dependence and [D-Pen2,D-Pen5]enkephalin cross-tolerance in whole animals. An antinociceptive potentiation between morphine and DAMGO was observed in morphine-tolerant and -dependent mice whereas in naive animals the effects of the two drugs were additive.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Analgesics; Animals; Binding Sites; Corpus Striatum; Drug Interactions; Drug Tolerance; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Mice; Morphine; Morphine Dependence; Naloxone; Pain; Receptors, Opioid

The substituted glucopyranose ring structure 2-hydroxypropyl-beta-cyclodextrin (CDEX) increases the solubility of molecules by inclusion of the agent in the lipophilic interior of the ring. This property is of particular use for the administration of molecules by the intracerebral (ICV) or intrathecal (IT) routes. In concentrations up to 40% w/v (isotonic), this agent (10 microliters) effect upon nociceptive or motor function after IT injection or on EEG and general behavior after ICV injection in rats. Using 20% CDEX, there is no change in the ED50 as compared to saline on the hot plate (HP) after IT injection of morphine, D-Ala2-D-Leu5 enkephalin or Tyr-Aib-Gly-gPhe-mAib-NH2, (Aib: alpha-aminoisobutyric acid) although there is an increase in their respective durations of effect. Cyclic peptide opioids: Tyr-c[D-A2bu-Gly-D-beta Nal(1)-D-Leu] (A2bu: alpha, gamma-diaminobutyric acid; beta-Nal(1): beta-naphthylalanine(1)) or Tyr-c[DA2bu-Gly-beta Nal(1)-D-Leu] are insoluble in saline but are readily dissolved in CDEX, and display a naloxone-sensitive antinociception following spinal administration. In other studies, saline insoluble capsaicin is administered in 25% dimethylsulfoxide (DMSO) or 20% CDEX (15 microliters; 5 mg/ml) which result in a significant reduction in the spinal levels of substance P and calcitonin gene related peptide and an increase in the HP latency. DMSO alone, but not CDEX alone, reduces the levels of the two peptides. These data emphasize the utility of complexation with CDEX for intracerebral drug delivery and compatibility with brain and spinal tissue.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; alpha-Cyclodextrins; Analgesia; Animals; beta-Cyclodextrins; Calcitonin Gene-Related Peptide; Capsaicin; Cerebral Ventricles; Cyclodextrins; Dimethyl Sulfoxide; Electroencephalography; Enkephalin, Leucine-2-Alanine; gamma-Cyclodextrins; Injections, Intraventricular; Injections, Spinal; Male; Morphine; Motor Activity; Naloxone; Narcotics; Pain; Pharmaceutical Vehicles; Rats; Rats, Inbred Strains; Solubility; Substance P; Time Factors

Some D-amino acid-substituted derivatives of bovine beta-casomorphin1-5 (CM) were found to share a particular affinity to central opioid receptors. As far as the antinociceptive potency of CM derivatives and their ability to produce tolerance is concerned, possible interactions of two selected CMs (Tyr-Pro-Phe-D-Pro-Gly, D-Pro4 and Tyr-Pro-D-Phe-Pro-Gly, D-Phe3) with morphine and, moreover, the doses necessary to induce tolerance were studied. Antinociception was tested by using the hot-plate test in experiments with mice. It was found that both CMs injected in doses up to 1 mumol/kg were not able to produce any sign of antinociception. It is all the more so interesting since such non-antinociceptive doses of D-Phe3 decreased analgesia induced by 15 mumol/kg morphine significantly. This could be regarded as a consequence of delta-receptor occupation by D-Phe3 and a resulting shift in the mu-receptor-mediated signal transduction. Finally, we found that repeated applications (once daily over a period of 20 days) induced a detectable tolerance to morphine. This suggests that opioid analgesia and tolerance must not be mediated by the same mechanisms and, moreover, that the mechanisms responsible for the development of tolerance are able to react more sensitive to an opioid.

Topics: Amino Acid Sequence; Animals; Drug Interactions; Endorphins; Enkephalin, Leucine-2-Alanine; Male; Mice; Mice, Inbred ICR; Molecular Sequence Data; Morphine; Pain; Phenylalanine; Proline; Stereoisomerism; Structure-Activity Relationship

Cholera toxin, an agent that impairs the function of Gs transducer proteins, was injected (0.5 microgram/mouse, icv) and the antinociceptive activity of opioids and clonidine was studied 24h later in the tail-flick test. In these animals, an enhancement of the analgesic potency of morphine, beta-endorphin and clonidine could be observed. Cholera toxin did not modify the antinociception evoked by the enkephalin derivatives DAGO and DADLE. Pertussis toxin that catalyses the ADP ribosylation of alpha subunits of Gi/Go regulatory proteins was given icv (0.5 microgram/mouse). This treatment reduced the analgesic effect of opioids and clonidine. However, while the analgesia elicited by DAGO, DADLE and clonidine was greatly decreased, the effect of morphine and beta-endorphin was reduced to a moderate extent. It is concluded that Gi/Go regulatory proteins functionally coupled to opioid and alpha 2 receptors are implicated in the efficacy displayed by opioids and clonidine to produce supraspinal analgesia. Moreover, these two receptors are susceptible to regulation by a process that might involve a Gs protein.

Topics: Analgesia; Animals; beta-Endorphin; Cerebral Ventricles; Cholera Toxin; Clonidine; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; GTP-Binding Proteins; Injections, Intraventricular; Male; Mice; Morphine; Narcotics; Pain; Pertussis Toxin; Receptors, Adrenergic, alpha; Virulence Factors, Bordetella

alpha N-acetyl human beta-endorphin-(1-31) injected icv to mice antagonized the analgesic activity of beta-endorphin-(1-31) and morphine whereas the analgesia evoked by DADLE and DAGO was enhanced by this treatment. The modulatory activity of alpha N-acetyl beta-endorphin-(1-31) was exhibited at remarkable low doses (fmols) reaching a maximum that persisted even though the dose was increased 100,000 times. The regulatory effect of a single dose of the acetylated neuropeptide lasted for 24h. The activity of alpha N-acetyl human beta-endorphin-(1-31) was partially retained by the shorter peptide alpha N-acetyl human beta-endorphin-(1-27) and to a lesser extent by beta-endorphin-(1-27), beta-endorphin-(1-31) lacked this regulatory activity on opioid analgesia. Acetylated beta-endorphin-(1-31) displayed a biphasic curve when competing with 5 pM [125I]-Tyr27 human beta-endorphin-(1-31) specific binding, the first step (20 to 30% of the binding) was abolished with an apparent IC50 of 0.35 nM, and the rest with an IC50 of 200 nM. It is suggested that alpha N-acetyl beta-endorphin-(1-31) changed the efficiency of the opioid analgesics by acting upon a specific substrate that is functionally coupled to the opioid receptor, presumably the guanine nucleotide binding regulatory proteins Gi/Go.

Topics: Acetylation; Animals; beta-Endorphin; Dose-Response Relationship, Drug; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Mice; Pain; Structure-Activity Relationship; Tail; Time Factors

Mice treated chronically with opioid antagonists have increased receptor density in brain and are supersensitive to the pharmacodynamic action of morphine. In the present study mice were implanted subcutaneously with naltrexone or placebo pellets for 8 days. During implantation mice received daily injections of morphine (100 or 250 mg/kg) or saline. Morphine analgesia was completely blocked in mice that were implanted with naltrexone at the low dose of morphine; while some analgesic action was observed at the higher dose. Mice implanted with placebo were analgesic following the daily morphine treatment. At the end of 8 days the pellets were removed and 24 h later some mice were tested for morphine analgesia while others were examined in binding studies. Naltrexone treatment increased [3H]naloxone, 3H[D-Ala2-D-Leu5]enkephalin (DADLE) and 3H[D-Ala2,NMePhe4,Gly-ol5]enkephalin (DAGO) binding compared to controls and increased the analgesic potency of morphine. Daily treatment with morphine did not alter brain opioid binding or naltrexone-induced receptor upregulation. Mice injected daily with morphine were significantly less sensitive to morphine (tolerant) than their respective saline control group for both the placebo and the naltrexone-treated groups. However, naltrexone-treated mice were more sensitive to morphine than placebo controls regardless of whether they were injected daily with morphine or not. These results indicate that if naltrexone-induced opioid receptor upregulation occurs in the presence of repeated agonist administration, the new binding sites mediate tolerance via desensitization to morphine.

Topics: Analgesia; Animals; Brain; Drug Tolerance; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine-2-Alanine; Enkephalins; Kinetics; Male; Mice; Morphine; Naloxone; Naltrexone; Pain; Receptors, Opioid; Reference Values; Up-Regulation

The antinociceptive effect of dalargin was studied by its influence on the amplitude of somatosensory induced potentials (IP) and by changes in the threshold pain sensitivity to heat stimulation. It was shown that it was necessary to taken into account the patients' individual sensitivity to the drug. The influence of dalargin on IP in the pre-operative period was compared with the quality of its analgetic defence in surgical interventions. The defence quality was determined according to the reduction in the consumption of the narcotic analgesic phentanyl during anesthesia. The studied parameters were found to be directly related to the correlation coefficient 0.75 (p less than 0.05). The findings allow for a reliable prediction of the antinociceptive effect of dalargin in its application in anaesthesiological practice.

Topics: Adult; Analgesics; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Evoked Potentials, Somatosensory; Humans; Middle Aged; Pain; Preanesthetic Medication; Sensory Thresholds

Several penicillins have been found to have pro-antinociceptive properties and also to be enkephalinase (neutral endopeptidase-24.11) inhibitors, carfecillin being the most potent. Carfecillin i.c.v. (but not i.p.) had significant antinociceptive activity in the mouse tail immersion test and completely suppressed abdominal constrictions (acetic acid) in mice (IC50 = 23 micrograms/animal). In combination with (D-Ala2-D-leu5)-enkephalin (DADL) i.c.v. in the abdominal constriction test the complete protection observed was reversed by the opioid receptor antagonist naltrexone. Carfecillin was a competitive inhibitor of enkephalinase from mouse brain striata (IC50 = 207 + 57 nM, cf thiorphan 10.6 +/- 1.9 nM) but did not inhibit other known enkephalin- degrading enzymes. Carfecillin provides a new lead structure for the development of more potent enkephalinase inhibitors.

Topics: Analgesics; Animals; Brain; Enkephalin, Leucine-2-Alanine; Isoenzymes; Male; Mice; Mice, Inbred Strains; Naltrexone; Neprilysin; Pain; Penicillins; Thiorphan; Time Factors

By employing a constant-rate (1 microliter/hr) constant-dose intrathecal infusion pump in rats, groups received a 7-day infusion of one of three concentrations (in nmol/hr) of each of the following receptor preferring ligands: morphine (mu: 2, 6, 20), sufentanil (mu: 0.06, 0.2, 0.6), DAMGO (mu: 0.1, 0.3, 1.0), DADLE (delta: 2, 6, 20) or ST-91 (alpha-2: 3, 10, 30). On day 7 the magnitude of tolerance was assessed by establishing i.t. dose-response curves for the effect of the chronic drug given as a bolus. Each infused animal was used for one bolus injection. It was observed that for all drugs, a parallel rightward shift in the dose-response curve was produced with the degree of shift proportional to the log of the infusion dose. Of particular significance was the finding that drugs differed significantly in the magnitude of the shift produced by the chronic infusion of equieffective concentrations. Thus, at the intermediate infusion dose, the degree of the shift in the bolus dose-response curve (i.e., magnitude of tolerance) for morphine, DADLE, ST-91, DAMGO and sufentanil was 55, 46, 29, 7 and 2, respectively. These data are interpreted to suggest that agents with high potency and a significant receptor reserve will produce a smaller shift in the dose-response curve (e.g., sufentanil and DAMGO) than the similar infusion of an equieffective dose of another agent with less potency and a smaller population of spare receptors (morphine).

Topics: Analgesics; Animals; Clonidine; Drug Tolerance; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Fentanyl; Infusions, Parenteral; Male; Morphine; Pain; Rats; Rats, Inbred Strains; Reference Values; Sufentanil

It has been demonstrated in experiments on unrestrained and unanesthetized curarized cats that periaqueductal gray matter stimulation produce sympathetic-activating action, raise arterial pressure and heart rate, but at the same time is not effective enough to suppress the nociceptive shifts of haemodynamic reactions. Opioid mechanisms of spinal cord plays an essential role in sympathetic-activating action of periaqueductal gray matter. It is suggested that the influence of antinociceptive areas of the brain stem on sympathetic haemodynamic regulation is one of the causes of resistance of nociceptive haemodynamic reactions to narcotic analgetics.

Topics: Animals; Blood Pressure; Cats; Electric Stimulation; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Kidney; Morphine; p-Chloroamphetamine; Pain; Periaqueductal Gray; Receptors, Opioid; Serotonin; Sympathetic Nervous System

Chronic treatment with opioid antagonists increases the potency of opioid agonists and produces an increase in brain opioid binding sites. In the present study, 8 day treatment with naltrexone blocked morphine and DADLE analgesia for the entire treatment period and increased mu 1, mu 2 and delta opioid receptor binding sites in mouse brain. mu 1 and mu 2 binding were increased by 81 and 67%, respectively, while delta binding was increased by 31%. Consistent with these binding changes, the potency of ICV morphine to produce analgesia was increased by over 3-fold, while the potency of ICV DADLE was increased by only 1.7. These findings indicate that relative increases in opioid receptor subtypes agree with pharmacodynamic studies on potency changes of opioid agonists.

Topics: Animals; Brain; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Male; Mice; Morphine; Naltrexone; Pain; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu

Bilateral intranigral microinjection of morphine produced dose-related and naloxone-reversible antinociceptive effects on the tail-flick and hot-plate tests. Intranigral injection of enkephalin had antinociceptive effects on both tests, and dynorphin had an antinociceptive effect on the hot-plate test. This is the first report of evidence that nigral opiate receptors may mediate antinociception.

Topics: Animals; Dynorphins; Endorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Microinjections; Morphine; Naloxone; Pain; Pain Measurement; Rats; Rats, Inbred Strains; Reaction Time; Substantia Nigra

Large baseline differences were found in open field activity and rearing behavior between 2 psychogenetically selected rat lines, as well as differences in the effects of peripherally administered morphine (2,4 and 10 mg/kg) on those behaviors and on body temperature. The 2 lines also showed different capacities of tritiated DAGO binding to preparations of striatal membranes.

Topics: Animals; Body Temperature; Cell Membrane; Corpus Striatum; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Morphine; Motor Activity; Pain; Rats; Rats, Mutant Strains; Receptors, Opioid; Species Specificity

Topics: Animals; Dynorphins; Endorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Female; Injections, Spinal; Morphine; Pain; Rats; Rats, Inbred Strains; Reflex; Sensory Thresholds; Urination

Dynorphin and [D-ala2-D-leu]enkephalin (DADLE) were administered directly into the cerebrolateral ventricles of rats and effects on various indices of sensorimotor function and retention of a passive avoidance task were measured. Dynorphin markedly suppressed exploratory motor activity and decreased responsiveness to an acoustic stimulus. Although increases in latency to respond to a noxious thermal stimulus were seen in rats after dynorphin, these changes were always associated with alterations in motor capacity. Injection of dynorphin immediately after a passive avoidance training trial had no significant effect on retention 1 week later. The physiological effects of DADLE were clearly different than those of dynorphin. DADLE produced a biphasic decrease followed by an increase in motor activity and an increased acoustic startle reactivity. DADLE had no effect on reactivity to a noxious thermal stimulus. Posttrial administration of DADLE significantly impaired retention of a step-through passive avoidance task 1 week after training. These data indicate different neurobiological roles for kappa and delta opiate receptors in the central nervous system.

Topics: Acoustic Stimulation; Animals; Avoidance Learning; Cerebral Ventricles; Dynorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Hot Temperature; Injections, Intraventricular; Male; Motor Activity; Pain; Rats; Rats, Inbred F344; Reflex, Startle

Stress (e.g. restraint) potentiates analgesia and alters changes in body temperature induced by morphine administered either systemically or intracerebroventricularly (i.c.v.) in rats. In order to extend the generality of this phenomenon to opioid peptides, we determined whether the analgesic and thermic effects of i.c.v. D-Ala2-D-Leu5-enkephalin (DADLE) or D-Ala2-N-MePhe4-Gly5(ol)-enkephalin (DAGO), agonists selective for delta- and mu-opioid receptors, respectively, were affected by restraint stress. Analgesia was measured in the tail-flick test and core body temperature by rectal probe. The unstressed rats exhibited a dose-dependent increase in tail-flick latencies after administration of either DAGO or DADLE. Restrained rats treated with DAGO or DADLE had a greater analgesic response to each dose of peptide than did unstressed rats; both the magnitude and duration of the drug effect were increased. The unstressed group of rats responded to all doses of DAGO and DADLE with an increase of core temperature. In contrast, restrained rats showed a decrease of core temperature following injection with either DAGO or DADLE. Thus, restraint stress can significantly modify the effects of DAGO and DADLE on analgesia and body temperature in a manner that is qualitatively and quantitatively similar to that observed previously for morphine administered by the i.c.v. route.

Topics: Animals; Body Temperature; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Injections, Intraventricular; Male; Morphine; Pain; Rats; Rats, Inbred Strains; Receptors, Opioid; Restraint, Physical; Stress, Physiological

The behavioural disturbances produced by acute exposure to ethanol have been related to changes in function of the opioid systems in the CNS. However, evidence in the literature is conflicting. The present report concerns the possible role of the enkephalinergic system in the mediation of acute ethanol effects. We used rats to study the ability of a selective opioid delta receptor antagonist (ICI 154129) to prevent the effect of ethanol on pain sensitivity, body temperature, sensorimotor performance and level of consciousness. Furthermore, in vitro receptor binding was measured to investigate whether or not ethanol, within a non-lethal concentration range, would change the binding parameters of the delta receptor ligand [3H][D-ala2, D-Leu5]enkephalin. ICI 154129 did not significantly influence the effects of ethanol in the behavioural experiments. Ethanol did not significantly change the binding parameters whether saturation or competition was measured in the receptor binding experiments. Thus, there was no evidence that the enkephalinergic system mediated the acute ethanol effects.

Topics: Animals; Behavior, Animal; Body Temperature; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Ethanol; In Vitro Techniques; Narcotic Antagonists; Pain; Psychomotor Performance; Rats; Rats, Inbred Strains; Reaction Time; Receptors, Opioid

Bilateral microinjections of DADL (D-Ala2-D-Leu5-enkephalin) and morphine were carried out in rats in a systematic fashion at histologically identified medial and lateral thalamic sites. DADL produced a dose-dependent (1.5-15.0 nmol), naloxone-reversible (1 mg/kg, i.p.) increase in the hot-plate (HP), tail-flick (TF) and catalepsy (CAT) response latencies with a predominance of activity occurring at lateral as opposed to medial thalamic sites. These effects were seen within 5 min of microinjection. At a significant number of sites, DADL precipitated convulsive seizure activity. Equimolar doses of morphine had a negligible effect on nociceptive indices and were not productive of seizures even at sites where DADL was found to be active. To further examine seizure activity, rats were prepared with bilateral frontal cortical electrodes and microinjected also at medial and lateral thalamic sites with equimolar doses of DADL and morphine (15 nmol). DADL was found to produce electrographically defined seizures unaccompanied by convulsive motor behavior (cataleptic seizures), as well as convulsive seizures. All animals in this group exhibiting analgesia and catalepsy had electrographic evidence of a seizure with markedly abnormal EEG tracings showing postictal spiking and changes in baseline frequency and amplitude. These seizures appeared to be naloxone-reversible. Morphine on the other hand was not productive of seizures, but did produce changes in electroencephalographic activity including spindle bursting, high-voltage slow-frequency activity as well as spiking. As noted, these changes were not associated with any effects on nociceptive measures.

Topics: Animals; Electroencephalography; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Male; Morphine; Pain; Rats; Rats, Inbred Strains; Receptors, Opioid; Receptors, Opioid, delta; Seizures; Sensory Thresholds; Thalamus

The analgesic activity of the prototypic opioid peptides for the mu (D-Ala2-Me-Phen4-Gly-ol5-enkephalin [DAGO]) kappa (Dynorphin 1-13), delta (D-Ala2-D-Leu5-enkephalin [DADLE]), or epsilon (beta-endorphin) receptor was assessed in a rat tooth pulp stimulation procedure. All opioid peptides tested and the opioid alkaloid U50, 488H (kappa receptor agonist) significantly elevated response thresholds. The rank order of potency based on the Minimum Effective Dose values was beta-endorphin greater than DAGO = dynorphin A (1-13) amide greater than DADLE greater than dynorphin A (1-13) greater than U50,488H. Based on absolute magnitude, the rank order of dose response slopes was DAGO greater than U50,488H greater than dynorphin A (1-13) amide greater than beta-endorphin greater than DADLE. Dynorphin A (1-13) produced the shallowest dose response slope and the magnitude of response threshold was the lowest for all compounds tested. Finally, the general conclusion that mu agonists are effective against noxious stimuli derived from thermal, chemical, and mechanical is extended by our data to include electrical sources derived from tooth pulp stimulation; kappa agonists are effective against noxious stimuli derived from chemical, mechanical, and electrical sources (tooth pulp stimulation) and delta agonists are effective analgesics against thermal, chemical and electrical stimuli (tooth pulp stimulation).

Topics: 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer; Analgesics; Animals; beta-Endorphin; Dental Pulp; Dynorphins; Electric Stimulation; Endorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Male; Pain; Peptide Fragments; Pyrrolidines; Rats; Rats, Inbred Strains

The analgesic effect of morphine in the rat tail jerk assay was enhanced by the serotonin uptake inhibitor, fluoxetine. Tail jerk latency was not affected by fluoxetine alone. Morphine's affinity for opioid receptors labeled in vitro with 3H-naloxone or 3H-D-Ala2-D-Leu5-enkephalin was not altered by fluoxetine, which has no affinity for these sites at concentrations as high as 1000 nM. In rats trained to discriminate morphine from saline, fluoxetine at doses of 5 or 10 mg/kg were recognized as saline. Increasing the fluoxetine dose to 20 mg/kg did not result in generalization to either saline or morphine. The dose response curve for morphine generalization was not significantly altered by fluoxetine doses of 5 or 10 mg/kg. Those rats treated with the combination of morphine and 20 mg/kg of fluoxetine did not exhibit saline or morphine appropriate responding. Fluoxetine potentiates the analgesic properties of morphine without enhancing its affinity for opioid receptors or its discriminative stimulus properties.

Topics: Animals; Binding, Competitive; Brain; Discrimination, Psychological; Drug Synergism; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Female; Fluoxetine; Male; Morphine; Naloxone; Pain; Propylamines; Rats; Rats, Inbred Strains; Receptors, Opioid; Serotonin

Implantation of rats with prolonged-release naltrexone pellets increased both morphine's analgesic potency in the tailflick assay and radiolabeled opioid binding in the brain. The increases in both radiolabeled opioid binding and morphine potency were time-dependent. Implantation for 24 hours did not increase binding, whereas increases of approximately 45% were seen following 8 days of implantation. Similarly, morphine's analgesic potency, measured as ED50 values, was increased by 50% following 8 days of exposure to naltrexone while a 24 hour exposure had no significant effect.

Topics: Animals; Binding, Competitive; Brain; Drug Implants; Drug Interactions; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Male; Morphine; Naloxone; Naltrexone; Pain; Rats; Rats, Inbred Strains; Reaction Time; Receptors, Opioid; Time Factors

D-Ala2-D-Leu5-enkephalin (DADL) along with dimers formed from tetra(DTE: des-Leu5-enkephalin)- or pentapeptide (DPE: Leu5-enkephalin) coupled by methylene bridges of various lengths (n) have been shown in in vitro systems to possess varying degrees of mu/delta-receptor selectivity. In the present studies we have systematically compared the intrathecal effect of these agents and morphine on the cutaneous stimuli (hot plate (HP) and tail flick (TF) and visceral-chemical (writhing) tests in the rat. The following observations were made. (1) Dimers with high delta-receptor selectivity were active in the TF(DPE2 greater than or equal to DADL greater than or equal to DTE2 greater than or equal to morphine greater than DPE12 much greater than DTE12 = 0) and HP(DPE2 greater than or equal to DTE2 greater than or equal to DADL greater than or equal to morphine greater than DPE12 greater than or equal to DTE12 greater than 0. To examine cross-tolerance, the intrathecal ED50 for morphine, DPE and DADL were determined in rats rendered tolerant by subcutaneous morphine pellets. The TF ED50 tolerant/TF ED50 naive was 18.4, 5.4 and 1.3, respectively. The ratio of activity on the HP was 14.0, 4.7 and 2.2 (2) On the visceral-chemical test, only morphine was active. The dimers or DADL in doses which totally blocked the TF or HP are at higher doses which were just below those producing motor dysfunction had no effect on the writhing response. (3) At high intrathecal doses (40 X TF ED50), morphine produced a motor rigidity which blocked the placing and stepping reflex.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acetates; Acetic Acid; Analgesics; Animals; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Hot Temperature; Male; Morphine; Pain; Rats; Rats, Inbred Strains; Receptors, Opioid; Skin; Spinal Cord

We have developed a quantitative computerized subtraction technique to demonstrate in rat brain the regional distribution of mu1 sites, a common very-high-affinity binding site for both morphine and the enkephalins. Low concentrations of [D-Ala2, D-Leu5]enkephalin selectively inhibit the mu1 binding of [3H]dihydromorphine, leaving mu2 sites, while low morphine concentrations eliminate the mu1 binding of [3H][D-Ala2, D-Leu5]enkephalin, leaving delta sites. Thus, quantitative differences between images of sections incubated in the presence and absence of these low concentrations of unlabeled opioid represent mu1 binding sites. The regional distributions of mu1 sites labeled with [3H]dihydromorphine were quite similar to those determined by using [3H][D-Ala2, D-Leu5]enkephalin. High levels of mu1 binding were observed in the periaqueductal gray, medial thalamus, and median raphe, consistent with the previously described role of mu1 sites in analgesia. Other regions with high levels of mu1 binding include the nucleus accumbens, the clusters and subcallosal streak of the striatum, hypothalamus, medial habenula, and the medial septum/diagonal band region. The proportion of total specific binding corresponding to mu1 sites varied among the regions, ranging from 14% to 75% for [3H][D-Ala2, D-Leu5]enkephalin and 20% to 52% for [3H]dihydromorphine.

Topics: Animals; Autoradiography; Binding Sites; Brain Chemistry; Dihydromorphine; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Pain; Periaqueductal Gray; Raphe Nuclei; Rats; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, mu; Subtraction Technique; Thalamus

The antinociceptive properties of morphine (mu), ethylketazocine (kappa), nalorphine (kappa), [D-Ala2, D-Leu5]enkephalin (delta) and [D-Ala2, Met5]enkephalinamide (mu, delta) were assessed using the radiant heat tail-flick and acetic acid-induced writhing assays in mice. The apparent pA2 values for the interaction of naloxone with morphine were the same regardless of the nociceptive stimulus employed or the route of administration of morphine. The apparent pA2 values for the interactions of naloxone with ethylketazocine and nalorphine in the writhing test differed significantly from that for the interaction of morphine and naloxone. Nalorphine did not produce a consistent antinociceptive effect on the tail-flick test. The apparent pA2 values for the interaction of ethylketazocine (s.c. or i.c.v.) with naloxone were similar to those for morphine-naloxone interactions on the tail-flick assay. The apparent pA2 values for the interactions of naloxone with [D-Ala2, D-Leu5] enkephalin differed from those for morphine-naloxone interactions on the writhing test. The highly selective mu antagonist beta-funaltrexamine antagonized the agonist actions of morphine and [D-Ala2, D-Leu5]enkephalin, and, in a previous study, beta-funaltrexamine antagonized the antinociceptive actions of [D-Ala2, Met5]enkephalinamide, but not those of nalorphine. It was concluded that agonist interaction with mu or kappa receptors can result in antinociceptive effects in the acetic acid-induced writhing test, and that an agonist interaction with mu, but not kappa, receptors results in antinociceptive action on the radiant heat tail-flick test, and furthermore, that a possible combination of mu and delta receptor interaction can result in antinociceptive activity in both tests.

Topics: Acetates; Acetic Acid; Analgesics, Opioid; Animals; Dose-Response Relationship, Drug; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Female; Hot Temperature; Male; Mice; Naloxone; Naltrexone; Pain; Receptors, Opioid; Receptors, Opioid, delta; Receptors, Opioid, kappa; Receptors, Opioid, mu

In rats implanted with chronic catheters in the spinal subarachnoid space, intrathecal injections of SKF 10047 and dynorphin did not produce any elevation of the nociceptive threshold as defined by hot-plate and tail-flick tests. In contrast, intrathecal ethylketocyclazocine (EKC) and (D-Ala2,D-Leu5)-enkephalin (DADL) administration resulted in a dose-dependent antinociceptive effect which was reversible with intraperitoneal naloxone. Calculation of the Schild dose-ratio plots for the data derived from systemically administered naloxone reveals a slope of--1 and a calculated pA2 value of 6.8 for EKC and 6.2 for DADL. Also, animals made tolerant to systemic morphine showed a diminished analgesic response to intrathecal morphine and EKC when compared to naive animals. There was, however, no significant change in the dose response curve of intrathecal DADL. Thus, these experiments suggest that in addition to mu receptors a separate subpopulation of delta but not kappa or sigma receptors are involved with spinally mediated analgesia.

Topics: Animals; Cyclazocine; Dose-Response Relationship, Drug; Dynorphins; Endorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Ethylketocyclazocine; Injections, Spinal; Male; Naloxone; Pain; Phenazocine; Rats; Rats, Inbred Strains; Receptors, Opioid; Spinal Cord

The systemic administration of subanalgesic doses of [D-Ala2, D-Leu5]enkephalin significantly potentiated the analgesia elicited in rats or mice by intraventricular injection of substance P. On the contrary, systemic administration of low doses of [D-Ala2,Met5]enkephalinamide antagonized the substance P-induced analgesia. The results support the notion of different physiological functions for the enkephalins and suggest an integrated role for enkephalins and substance P in the control of pain at supraspinal levels.

Topics: Animals; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalin, Methionine; Injections, Intraventricular; Male; Mice; Mice, Inbred ICR; Pain; Rats; Rats, Inbred Strains; Substance P

Leu[Enkephalin (400.0 micrograms/kg) and the enkephalin analog [DAla,DLeu]enkephalin (0.4, 4.0, and 40.0 micrograms/kg) were intraperitoneally administered to rats 5 min before they were trained on aversively motivated tasks. The peptides impaired acquisition of a one-way active avoidance response, facilitated acquisition of an inhibitory avoidance response, and had no effect on acquisition of a swim-escape response. The data indicate that the enkephalin effects are not mediated through actions on locomotor activity, pain perception or reactivity, or light sensitivity. Rather, we suggest that [Leu]enkephalin and its analog strengthen the tendency of rats to suppress behavior in the presence of cues previously associated with aversive stimulation. This effect may be due to an enkephalin-induced increase in fear or arousal. The enkephalin effect on behavior was obtained at low doses and was observed within a few minutes after administration. These findings are consistent with the interpretation that enkephalin effects may be initiated at a peripheral site.

Topics: Animals; Avoidance Learning; Behavior, Animal; Electroshock; Endorphins; Enkephalin, Leucine; Enkephalin, Leucine-2-Alanine; Enkephalins; Escape Reaction; Fear; Light; Male; Pain; Perception; Rats; Sensory Thresholds; Visual Perception; Water