J. Cros

Autoradiographic localization of mu, delta and kappa opioid receptor binding sites in rat and guinea pig spinal cord

The autoradiographic distribution of mu, delta and kappa opioid binding sites was evaluated in various segments of the rat and guinea pig spinal cord. Mu opioid receptor binding sites are highly concentrated in the superficial layers of the dorsal horn (laminae II and III) in both species, without any marked gradient along the cord. Delta binding sites are somewhat concentrated in the superficial layers of the dorsal horn. However, delta binding sites are also present and evenly distributed in other areas of the gray matter. The highest density of delta sites is found in the cervical segment with only low levels in the lumbo-sacral region of the rat and guinea pig spinal cord. Kappa opioid binding sites are highly concentrated in the superficial layers of the dorsal horn of the spinal cord. Lower levels are seen in the rest of the gray matter with some enrichment in lamina X. Moreover, the lumbo-sacral portion of the spinal cord is enriched in kappa sites as compared to the cervical and thoracic segments. These data demonstrate the differential laminar distribution of mu, delta and kappa opioid binding sites in rat and guinea pig spinal cord.

Morphine analgesia and cerebral opiate receptors: a developmental study.

1 Development of the analgesic response to morphine and ontogenesis of central opiate receptors were analyzed in rats 5 to 120 days old. 2 The analgesic effect of morphine increased until day 15, after which it decreased to reach a plateau at about day 30. With phenoperidine, on the other hand, the analgesic effect increased until day 15, remained constant between day 15 and day 30 after which it decreased slowly. 3 The ratio of the amounts of morphine in blood over those in brain increased about 3 fold between day 15 and day 30. 4 Opiate receptors were detected in the brain of newborn rats: stereospecific binding of [3H]‐naloxone at 10 and 50 nM indicated the presence of low and high affinity binding sites. 5 The number of [3H]‐naloxone binding sites increased rapidly during the second and third week after birth. Their affinity for several opiates remained constant throughout development. 6 These results indicate that the analgesic activity of opiates varies with age: until day 15, the analgesic effect of opiates increases in parallel with the number of opiate brain receptors. Then, the formation of the blood brain barrier introduces an additional step in the regulation of opiate activity.

Opioid binding sites in different levels of rat spinal cord

Opioid receptor binding sites were analyzed in various segments of rat spinal cord. Mu and delta types were labelled with [3H]-DHM or [3H]-DAGO and [3H]-DADLE or [3H]-DSLET respectively. Kappa 1 (kappa) and kappa 2 (benzomorphan) binding sites were individually detected by the overall labeling of opioid binding sites with [3H]-etorphine followed by the elimination of binding to particular sites by the use of appropriate selective unlabelled ligands. Whereas lumbo sacral region contained mainly the kappa 2 site, thoracic membranes had a proportion of approximately 20% mu, 20% kappa 1 and 60% kappa 2 and cervical region contained much less kappa 2 sites (25% mu, 20% delta, 28% kappa 1 and 32% kappa 2).

CSF baclofen levels after intrathecal administration in severe spasticity.

SummaryThe pharmacokinetic parameters in the CSF of baclofen given to 4 patients as an intrathecal bolus are reported. Considerable inter-individual variability in the parameters was observed. The elimination half-life ranged from 0.9 to 5 h and the clearance from 0.013 to 0.08 l·h−1.In order to optimize treatment, it is suggested that CSF baclofen levels be matched to changes in Hoffmans monosynaptic reflex (H reflex).

Benzomorphan binding sites in rat lumbo-sacral spinal cord

The rat lumbo-sacral spinal cord contains a homogeneous population of opiate binding sites labelled with high affinity (KD = 0.21 +/- 0.04 nM) by [3H]etorphine and lower affinity (KD = 2.2 +/- 0.4 nM) by [3H]ethylketocyclazocine. Benzomorphan drugs are potent competitors for these binding sites while morphine and enkephalin display a low affinity. These binding sites have binding properties which are distinct from the mu-, delta-, and also kappa-sites but are very similar to those of the benzomorphan sites characterized in rat brain.

Specific binding for opiate-like drugs in the placenta

Abstract The properties of human placental opiate binding sites were analysed using [3H]etorphine in terms of kinetic parameters, specificity, subcellular distribution and effect of ionic environment. In vitro these sites display a pharmacological profile similar to that of the pituitary opiate receptor site. In particular, they exhibit lower affinities for opioid peptides than do brain opiate receptors. They were detected exclusively in human placenta, in which they appeared during the first half of pregnancy.

Purification and identification of multiple forms of dynorphin in human placenta.

Immunoreactive dynorphin (ir-Dyn) and opiate-like peptides (OLP) were measured in acid (HC1) extracts of human placenta by the use of an antibody to synthetic Dyn-(1-13) and of the displacement of [3H]-naloxone binding to rat brain homogenates, respectively. The placenta contained 57.6 pmoles per g of ir-Dyn and 134.4 pmoles per g of naloxone binding equivalents. After passage of the extract through cartridges of Sep Pak C18, half of the OLP was eluted with ir-Dyn at 35% acetonitrile (ACN), the rest being eluted at 60% ACN. Both fractions obtained from Sep Pak were chromatographed separately on Sephadex G-50, the OLP of the 35% ACN fraction coeluting with the ir-Dyn speak and that of the 60% ACN fraction being eluted at the same volume as synthetic beta-endorphin. Conversely, the fraction of OLP coeluting with synthetic leucine-enkephalin (Leu-Enk) in these two chromatographies was minimal. The Dyn-immunoreactive material was further purified by high pressure liquid chromatography on reverse phase micro-Bondapak C18 columns to give three distinct peaks corresponding to synthetic Dyn-(1-11), Dyn-(1-13) and Dyn-(1-12), respectively. Our results indicate that the human placenta contains several forms of ir-Dyn which account for about half of its endogenous OLP.

Effects of ibogaine on naloxone‐precipitated withdrawal in morphine‐dependent mice

Summary— In naive mice, ibogaine at a tremorigenic dose (30 mg/kg, ip), did not produce antinociception but did potentiate the antinociceptive potency of morphine in the tail‐flick test. In morphine‐dependent mice, ibogaine did not eliminate withdrawal symptoms but significantly increased the number of repetitive vertical jumps induced by naloxone, whatever the duration of the chronic morphine treatment. By comparison, repetitive jumping induced by α‐napthoxyacetic acid (α‐NOAA), a non‐convulsant drug which induced jumping without affecting other morphine‐withdrawal signs, was not significantly modified by ibogaine. These results indicate that while acute antinociceptive effects of morphine are modulated by ibogaine, this drug, shown to alleviate opiate dependence in man, does not attenuate in mice opioid withdrawal manifestations.

Kappa opiate binding sites in human placenta.

Summary [ 3 H] etorphine, an agonist of μ, δ and κ receptors and [ 3 H] ethylketocyclazocine, a κ agonist, bind to one class of binding sites with a same maximum number of binding sites on human placental membranes. The pharmacological characteristics and the sensitivity to sodium ions are strictly identical for both ligands and correspond to the pharmacological properties of the κ receptor. This is also confirmed by the absence of detectable specific binding for [ 3 H]-(DAla) 2 -(DLeu) 5 enkephalin, a δ agonist and [ 3 H]DHM, a μ agonist. Human placental membranes can therefore be considered as a tissue containing a homogenous population of κ opiate binding sites.

Kappa opioid receptors in human lumbo-sacral spinal cord.

[3H]Etorphine and [3H]ethylketocyclazocine bind with high affinity (Kd between 0.25-2.0 nM) to a single class of sites in human lumbo-sacral spinal cord. Other ligands such as [3H]morphine, [3H]dihydromorphine and [3H]D-Ala2, D-Leu5-enkephalin (DADLE) did not bind to significant number of sites under our incubation conditions. Ligand selectivity pattern strongly suggests that [3H]etorphine labels kappa opioid binding sites in the human lumbo-sacral spinal cord since benzomorphans and oripavines are much more potent than mu and delta agonists. Furthermore, [3H]etorphine and [3H]ethylketocyclazocine binding is sensitive to high concentrations of DADLE suggesting that these sites are of the kappa 2 sub-type. Finally, the visualization of these sites by receptor autoradiography demonstrates that they are mainly concentrated in lamina II and III of the dorsal horn. Moderate densities of sites are present around the central canal. Thus, it is possible that kappa opioid binding sites could be involved in the control of sensory and autonomic functions in the human lumbo-sacral spinal cord.