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  • 1
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: This study was carried out to analyze the extracellular levels of Met-enkephalin-like material in the nucleus accumbens, a brain structure involved in the effects of opioids on motor activity and reward processes, using microdialysis in awake and freely moving rats, combined with a sensitive radioimmunoassay. The levels of Met-enkephalin-like material were measured after administration of a dual inhibitor of enkephalin-degrading enzymes, RB101, to evaluate its in vivo protecting effects. The basal levels of Met-enkephalin-like immunoreactivity in the nucleus accumbens were ∼1.2 pg/30 min or 2.2 fmol/30 min (37 pM). Perfusion of KCl (100 mM) produced a 17-fold increase in the level of Met-enkephalin-like material in this structure. During the 8-h perfusion, which started at 9 a.m., a spontaneous increase of the basal level of Met-enkephalin-like material in the nucleus accumbens occurred between 4 and 4:30 p.m., suggesting the existence of variation in opioid peptide secretion, at least in this structure. Intraperitoneal injection of RB101 induced a dose-dependent and long-lasting (210-min) increase in the extracellular levels of Met-enkephalin-like material. A prolonged effect was also observed in the behavioral studies in which the inhibitor increased global motor activity of rats 210 min after injection. These data represent the first direct evidence that dual inhibitors of enkephalin-degrading enzymes increase in vivo the extracellular levels of Met-enkephalin-like material in awake and freely moving rats.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Superfusion of slices from the dorsal half of the lumbar enlargement of rat spinal cord with Krebs-Henseleit medium supplemented with 30 μM bacitracin allowed the collection of substance P-like immunoreactive material (SPLI), which was released at a rate of ∼ 10 pg/4 min. Tissue depolarization by an excess of K+ (30–60 mM) or veratridine (50 μM) induced a marked increase in SPLI outflow, provided that Ca2+ was present in the superfusing fluid. K+-or veratridine-induced SPLI overflow could be modulated in opposite directions by μ and δ opioid receptor agonists. Thus, the two preferential μ agonists Tyr-d-Ala-Gly-MePhe-Gly-ol (DAGO; 10 μM) and Tyr-d-Ala-Gly-MePhe-Met(O)5-OH (FK-33824; 0.1 μM) enhanced SPLI overflow from depolarized tissues, whereas the selective δ agonists Tyr-d-Thr-Gly-Phe-Leu-Thr (deltakephalin; 3 μM) and [2-d-penicillamine, 5-d-penicillamine]enkephalin (50 μM) reduced it. The effect of DAGO was antagonized by a low concentration (1 μM) of naloxone but not by the selective δ antagonist ICI-154129 (50 μM). In contrast, the latter drug prevented the inhibitory influence of δ agonists on K+-induced SPLI release. Complementary experiments with morphine (10 μM) and [2-d-alanine, 5-d-leucine]enkephalinamide (3 μM), in combination with 1 μM naloxone or 50 μM ICI-154129 for the selective blockade of μ or δ receptors, respectively, confirmed that the stimulation of μ receptors increased, whereas the stimulation of δ receptors reduced, SPLI overflow. The results suggest that, at the spinal level, the antinociceptive action of δ but not μ agonists might involve a presynaptic inhibition of substance P-containing primary afferent fibers.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract : Because cholecystokinin (CCK) acts as a“functional” endogenous opioid antagonist, it has been proposedthat changes in central CCKergic neurotransmission might account for therelative resistance of neuropathic pain to the analgesic action of morphine.This hypothesis was addressed by measuring CCK-related parameters 2 weeksafter unilateral sciatic nerve section in rats. As expected, significantdecreases (-25-38%) in the tissue concentrations and in vitro release of bothsubstance P and calcitonin gene-related peptide were noted in the dorsalquadrant of the lumbar spinal cord on the lesioned side. In contrast, thetissue levels and in vitro release of CCK were unchanged in the same area inlesioned rats. Measurements in dorsal root ganglia at L4-L6 levels revealed nosignificant changes in proCCK mRNA after the lesion. However, sciatic nervesection was associated with a marked ipsilateral increase in both CCK-Breceptor mRNA levels in these ganglia (+70%) and the autoradiographic labelingof CCK-B receptors by [3H]pBC 264 (+160%) in the superficial layers of the lumbar dorsal horn. Up-regulation of CCK-B receptors rather than CCK synthesis and release probably contributes to increased spinal CCKergic neurotransmission in neuropathic pain.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Intrathecal infusion of the neuropeptide FF analogue, [D-Tyr1, (NMe)Phe3]neuropeptide FF (1DMe; 0.1 µm−0.1 mm) in anaesthetized rats produced a concentration-dependent decrease in the spinal outflow of dynorphin A (1–8)-like material, which persisted for at least 90 min after treatment with 10 µm−0.1 mm of the compound. Co-administration of d-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP; 1 µm) to block spinal µ-opioid receptors did not modify this effect, whereas naltrindole (10 µm) totally prevented it and nor-binaltorphimine (10 µm) reduced the post-effect. These data suggest that 1DMe triggers the release of endogenous opioids that stimulate mainly δ-opioid receptors, and secondarily κ-opioid receptors, thereby exerting a negative influence on dynorphin A (1–8)-like material outflow. Because dynorphin has pronociceptive properties, such a decrease in spinal dynorphin A (1–8)-like material release might underlie the long-lasting antinociceptive effects of intrathecally administered neuropeptide FF and analogues.
    Type of Medium: Electronic Resource
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