κ3 opiate receptor binding in the mouse and rat

doi:10.1016/0922-4106(92)90077-9

European Journal of Pharmacology: Molecular Pharmacology

Volume 226, Issue 1, 12 May 1992, Pages 15-20

https://doi.org/10.1016/0922-4106(92)90077-9 Get rights and content

Abstract

Previous work examining the binding of [³H]naloxone benzoylhydrazone (NalBzoH) in calf brain has identified a novel binding site, κ₃. In mice and rats NalBzoH elicits an analgesic response which can clearly be differentiated from classical μ, δ or κ₁ mechanisms and which is pharmacologically consistent with a κ₃ receptor mechanism of action. In the current studies we demonstrate the presence of κ₃ sites in both mouse and rat brains. The selectivity of the κ₃ sites for opioids clearly discriminated it from traditional μ, κ₁, κ₂ or δ receptors. In the rat, the density of κ₃ sites increased 2.5-fold from age 2 days to 21 days, after which it remained relatively stable. Among a number of brain regions in the rat, the density of κ₃ sites varied dramatically. Highest levels were present in the hypothalamus, thalamus, striatum and midbrain with very low levels in the cerebellum. Intermediate levels were present in cortex, brain stem and spinal cord. Together, these studies support the presence of κ₃ receptors in both mouse and rat brain which are very similar to those previously reported in calf brain.

References (37)

C. Burkhardt et al.
Metkephamid (TyrDAlaGlyPheN(Me)MetONH₂, a potent opioid peptide; receptor binding and analgesic properties
Peptides
(1982)
J.A. Clark et al.
U50,488, a κ-selective agent with poor affinity for mu₁ opiate binding sites
Neuropharmacology
(1988)
J.A. Clark et al.
[D-Pen², D-Pen⁵]Enkephalin (DPDPE): a delta-selective enkephalin with low affinity for μ₁ opiate binding sites
Eur. J. Pharmacol.
(1986)
V. Dauge et al.
Use of μ and δ opioid peptides of various selectivity gives further evidence of specific involvement of μ opioid receptors in supraspinal analgesia (tail flick test)
Eur. J. Pharmacol.
(1987)
D.W. Harris et al.
High affinity binding of [³H]ethylketocyclazocine to rat brain homogenate
Eur. J. Pharmacol.
(1980)
S. Iyengar et al.
Effects of kappa opiate agonists on neurochemical and neuroendocrine indices: evidence for kappa receptor subtypes
Life Sci.
(1986)
O.H. Lowry et al.
Protein measurement with the Folin phenol reagent
J. Biol. Chem.
(1951)
M.C. Luke et al.
Irreversible opiate agonists and antagonists. V. Novel hydrazone derivatives of naltrexone
Life Sci.
(1988)
B. Nock et al.
[³H]U-69593 labels a subtype of kappa opiate receptor with characteristics different from that labeled by [³H]ethylketocyclazocine
Life Sci.
(1988)
A. Pfeiffer et al.
Demonstration and distribution of an opiate binding site in rat brain with high affinity for ethylketocyclazocine and SKF10,047
Biochem. Biophys. Res. Commun.
(1981)

G.A. Weiland et al.

Mini review. Quantitative analysis of drug receptor interactions: (i) determination of kinetic and equilibrium properties

Life Sci.

(1981)

Y. Audiger et al.

Characterization of [³H]etorphine binding in guinea pig striatum after blockade of mu and delta sites

Life Sci.

(1982)

C. Chavkin et al.

Dynorphin is a specific endogenous ligand of the κ opioid receptor

Science

(1982)

Y.C. Cheng et al.

Relationship between the inhibition constant (K₁) and the concentration of inhibitor which causes 50 percent inhibition (IC₅₀) of an enzymatic reaction

Biochem. Pharmacol.

(1973)

T.C. Chou

Relationships between inhibition constants and fractional inhibition in enzyme-catalyzed reactions with different number of reactants, different reaction mechanisms and different types and mechanisms of inhibition

Mol. Pharmacol.

(1974)

J.A. Clark et al.

Opiate binding in calf thalamic membranes: a selective mu₁ binding assay

Mol. Pharmacol.

(1988)

J.A. Clark et al.

Kappa opiate receptor multiplicity: evidence for two U50,488-sensitive κ₁ subtypes and a novel κ₃ subtype

J. Pharmacol. Exp. Ther.

(1989)

H.A. Feldman

Mathematical theory of complex ligand binding systems at equilibrium: methods for parameter fitting

Ann. Biochem.

(1912)

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Increasing evidence suggests that the cerebellum could play a role in the higher cognitive processes involved in addiction as the cerebellum contains anatomical and functional pathways to circuitry controlling motivation and saliency. In addition, the cerebellum exhibits a widespread presence of receptors, including opioid receptors which are known to play a prominent role in synaptic and circuit mechanisms of plasticity associated with drug use and development of addiction to opioids and other drugs of abuse. Further, the presence of perineural nets (PNNs) in the cerebellum which contain proteins known to alter synaptic plasticity could contribute to addiction. The role the cerebellum plays in processes of addiction is likely complex, and could depend on the particular drug of abuse, the pattern of use, and the stage of the user within the addiction cycle. In this review, we discuss functional and structural modifications shown to be produced in the cerebellum by opioids that exhibit dependency-inducing properties which provide support for the conclusion that the cerebellum plays a role in addiction.
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The supraspinal and spinal antinociceptive effects of several κ-opioid receptor agonists were examined in diabetic and non-diabetic mice using the tail-flick assay. The antinociception induced by intrathecal (i.t.), but not intracerebroventricular (i.c.v.), CI-977, a highly selective κ₁-opioid receptor agonist, in diabetic mice was less than that in non-diabetic mice. The antinociceptive effects of ICI-199,441 and R-84760, high potency κ₁-opioid receptor agonists, given i.c.v., but not i.t., were attenuated in diabetic mice compared to those in nondiabetic mice. On the other hand, the antinociceptive effects of the new κ-opioid receptor agonist TRK-820, which has high affinity for κ₂- and/or κ₃-opioid receptors, injected both i.c.v. and i.t. in diabetic mice were markedly less than those in non-diabetic mice. These results indicate that the antinociceptive effects of those κ-opioid receptor agonists in diabetic mice are altered in a region-specific manner in the central nervous system (CNS). The dysfunction of κ-opioid receptor subtypes in diabetic mice may underlie this CNS region-specific variation in the effects of these κ-opioid receptor agonists.
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Citation Excerpt :
The early assays to assess binding to δ-opioid receptors were carried out with poorly selective ligands but these experiments suggested binding to δ-receptors was relatively weak (Price et al., 1988). Competition studies versus [3H]NalBzoH at 1 nM using δ-receptor ligands, showed a lack of effect of the peptide δ-agonist DPDPE but some affinity of the non-peptide δ-antagonist, naltrindole (Cheng et al., 1992). Although naltrindole has greater binding selectivity for the δ-receptor than DPDPE the contribution of δ-receptor activity is thus still unclear.
Naloxone benzoylhydrazone (NalBzoH) is a ligand used to study opioid receptors. It has been suggested to act at a novel kappa₃ receptor but also appears to bind to classical opioid receptors, and possibly the ORL1 receptor. We have used opioid receptor triple knockout mice, deficient in genes coding for the μ, δ and κ-receptor, to characterise the relative contributions of opioid and ORL1 activity to the binding of this ligand, by carrying out receptor autoradiography with [³H]NalBzoH. As competing ligands we have used diprenorphine and nociceptin at 1 μM, alone or in combination, to determine the contribution of opioid and ORL1 receptor binding. At 4 nM [³H]NalBzoH showed labelling in wild-type brains indicative of broad spectrum classical opioid receptor binding. In the triple knockout brains all labelling was completely absent, suggesting that at this concentration there is no binding to ORL1 sites. However at 50 nM [³H]NalBzoH showed labelling in triple knockout brains with a distribution pattern indicative of ORL1 labelling. Quantitative analysis showed that nociceptin displaced typically 30% of the residual labelling in knockout brains whilst diprenorphine had relatively little effect. The data show that at 50 nM NalBzoH no binding was detected other than to classical opioid receptors or to ORL1 in an approximate ratio of 2:1.
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Regular paperκ3 opiate receptor binding in the mouse and rat

Abstract

Peptides

Neuropharmacology

Eur. J. Pharmacol.

Eur. J. Pharmacol.

Eur. J. Pharmacol.

Life Sci.

J. Biol. Chem.

Life Sci.

Life Sci.

Biochem. Biophys. Res. Commun.

Life Sci.

Characterization of [3H]etorphine binding in guinea pig striatum after blockade of mu and delta sites

Life Sci.

Dynorphin is a specific endogenous ligand of the κ opioid receptor

Science

Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 percent inhibition (IC50) of an enzymatic reaction

Biochem. Pharmacol.

Relationships between inhibition constants and fractional inhibition in enzyme-catalyzed reactions with different number of reactants, different reaction mechanisms and different types and mechanisms of inhibition

Mol. Pharmacol.

Opiate binding in calf thalamic membranes: a selective mu1 binding assay

Mol. Pharmacol.

Kappa opiate receptor multiplicity: evidence for two U50,488-sensitive κ1 subtypes and a novel κ3 subtype

J. Pharmacol. Exp. Ther.

Mathematical theory of complex ligand binding systems at equilibrium: methods for parameter fitting

Ann. Biochem.

Regular paper
κ₃ opiate receptor binding in the mouse and rat

Characterization of [³H]etorphine binding in guinea pig striatum after blockade of mu and delta sites

Relationship between the inhibition constant (K₁) and the concentration of inhibitor which causes 50 percent inhibition (IC₅₀) of an enzymatic reaction

Opiate binding in calf thalamic membranes: a selective mu₁ binding assay

Kappa opiate receptor multiplicity: evidence for two U50,488-sensitive κ₁ subtypes and a novel κ₃ subtype