Autoradiographic analysis of mu1, mu2, and delta opioid binding in the central nervous system of C57BL/6BY and CXBK (opioid receptor-deficient) mice

doi:10.1016/0006-8993(85)91226-0

Brain Research

Volume 360, Issues 1–2, 23 December 1985, Pages 108-116

https://doi.org/10.1016/0006-8993(85)91226-0 Get rights and content

Abstract

The recent development of in vitro autoradiography techniques has enabled investigators to determine the distribution and relative levels of multiple ligand binding sites in discrete anatomical areas. In this study we used semi-quantitative in vitro autoradiography to compare the levels of binding to central mu₁, mu₂, and delta opioids sites in two strains of mice, C57BL/6BY and CXBK. The CXBK strain is known to be deficient in whole brain opioid binding sites and to be less sensitive than the C57 strain to the analgesic and locomotor stimulatory effects of opiates and opioids. Delta sites were visualized using [³H][d-Ala²-d-Leu⁵]-enkephalin (DADL) plus a low concentration of morphine, total mu sites (mu₁ and mu₂) were visualized using [³H]dihydromorphine (DHM), and mu₂ sites were visualized using [³H]DHM plus a low concentration of DADL. Binding to mu₁ sites was determined by subtracting mu₂ binding from total mu binding. We found that the two strains did not consistently differ in the levels of delta sites; in some areas the CXBKs had lower levels but in many areas they had levels equal to or greater than those for the C57s. The CXBK strain, however, either had less or the same amount of mu binding as the C57 strain in all areas studied. The CXBK strain was especially deficient in mu₁ binding, particularly in areas involved in pain processing.

Reference (60)

AtwehS.F. et al.
Autoradiographic localization of opiate receptors in rat brain. I. Spinal cord and lower medulla
Brain Research
(1977)
AtwehS.F. et al.
Autoradiographic localization of opiate receptors in rat brain. II. The brain stem
Brain Research
(1977)
AtwehS.F. et al.
Autoradiographic localization of opiate receptors in rat brain. III. The telencephalon
Brain Research
(1977)
BaranA. et al.
Opiate receptors in mice: genetic differences
Life Sci.
(1975)
ChengR.S.S. et al.
Correlation of genetic differences in endorphin systems with analgesic effects ofd-amino acids in mice
Brain Research
(1979)
DukaT. et al.
A selective distribution pattern of different opiate receptors in certain areas of rat brain as revealed by in vitro autoradiography
Neurosci. Lett.
(1981)
DukaT. et al.
Selective localization of different types of opiate receptors in hippocampus as revealed by in vitro autoradiography
Brain Research
(1981)
EdleyS.M. et al.
Evolution of striatal opiate receptors
Brain Research
(1982)
FooteR.W. et al.
Autoradiographic localization of opiate kappa receptors in the guinea-pig brain
Eur. J. Pharmacol.
(1982)
GoodmanR.R. et al.
Autoradiography of [³H]beta-endorphin binding in brain
Brain Research
(1983)

HughesJ.

Isolation of an endogenous compound from the brain with pharmacological properties similar to morphine

Brain Research

(1975)

MoskowitzA.S. et al.

Autoradiographic distribution of mu₁ and mu₂ opioid binding in the mouse central nervous system

Brain Research

(1985)

NeedhamW.P. et al.

Liver damage from narcotics in mice

Toxicol. Appl. Pharmacol.

(1981)

PasternakG.W. et al.

An endogenous morphine-like factor in mammalian brain

Life Sci.

(1975)

ReithM.E.A. et al.

Strain differences in opiate receptors in mouse brain

Eur. J. Pharmacol.

(1981)

ZhangA.-Z. et al.

Mu- and delta-opiate receptors: correlation with high and low affinity opiate binding sites

Eur. J. Pharmacol.

(1980)

ChangK.-J. et al.

Multiple opiate receptors: different regional distribution in the brain and differential binding of opiates and opioid peptides

Mol. Pharmacol.

(1979)

Della BellaD. et al.

Opiate receptors: different ligand affinity in various brain regions

FrenkH. et al.

Different brain areas mediate the analgesic and epileptic properties of enkephalins

Science

(1978)

GoldsteinA. et al.

Dynorphin-(1–13), an extraordinarily potent opioid peptide

GoodmanR.R. et al.

Kappa opiate receptors localized by autoradiography to deep layers of cerebral cortex: relation to sedative effects

GoodmanR.R. et al.

Differentiation of delta and mu opiate receptor localization by light microscopic autoradiography

HerkenhamM. et al.

In vitro autoradiography of opiate receptors in rat brain suggests loci of ‘opiatergic’ pathways

HerzA. et al.

Is there some indication from behavioral effects of endorphin for their involvement in psychiatric diseases?

HuangJ.T.

Study on morphine induced opacity in mouse lens: different age and strain

Res. Commun. Sub. Abuse

(1981)

HughesJ. et al.

Identification of two related pentapeptides from the brain with potent opiate agonist activity

Nature (London)

(1975)

JaffeJ.H.

Narcotic analgesics

KuharM.J. et al.

Regional distribution of opiate receptor binding in monkey and human brain

Nature (London)

(1973)

LeslieF.M. et al.

Opioid binding properties of brain and peripheral tissues: evidence for heterogeneity in opioid ligand binding sites

J. Pharmacol. Exp. Ther.

(1980)

LewisM.E. et al.

Opiate receptor gradients in monkey cerebral cortex: correspondence with sensory processing hierarchies

Science

(1981)

Cited by (125)

Receptors: Opioid receptors
2021, Encyclopedia of Biological Chemistry: Third Edition
Opioid receptor, a member of the superfamily of membrane proteins that transduces signals via the heterotrimeric G proteins, has myriad of roles upon activation by exogenous alkaloid and endogenous peptide ligands. The most important of these functions is the modulation of pain perception. The massive side effects associated with opioid receptor activation, especially those that can lead to addiction to the exogenous alkaloid ligands, mandate the investigation into the mechanism of opioid receptor action. In this article, we examine some of the opioid receptor signaling complexity.
5.35 - Forebrain Opiates
2020, The Senses: A Comprehensive Reference: Volume 1-7, Second Edition
This chapter reviews the current literature on supraspinal endogenous opioid systems and their functions in animal models and humans. The various opioid and opioid-like peptide families and receptors present a multiplicity of effects, with prominent, but not exclusive, effects on pain regulation. They are distributed in cortical and subcortical regions involved in sensory processing, as well as in higher order, integrative functions. Existing and emerging data in animal models and humans demonstrate the involvement of these neurotransmitters in sensory, emotional and cognitive processing, and the overlap between these functions.
Stress and Opioid Systems
2017, Hormones, Brain and Behavior: Third Edition
The benefits of magnetic resonance imaging methods to extend the knowledge of the anatomical organisation of the periaqueductal gray in mammals
2016, Journal of Chemical Neuroanatomy
Citation Excerpt :
Volume differences were observed between μ receptor-knockout and wild-type mice, especially in the vlPAG, which is bigger in the first ones. This latter result is consistent with the localisation of δ – opioid receptors in the mice vlPAG (Moskowitz and Goodman, 1985). Still in the aim to use in vivo methods, magnetic resonance spectroscopy could be used to describe the metabolites contain of brain structures (Chaillou et al., 2012).
The periaqueductal gray (PAG) is a mesencephalic brain structure involved in the expression of numerous behaviours such as maternal, sexual and emotional. Histological approaches showed the PAG is composed by subdivisions with specific cell organisation, neurochemical composition and connections with the rest of the brain. The comparison of studies performed in rodents and cats as the most often examined species, suggests that PAG organisation differs between mammals. However, we should also consider the plurality of the methods used in these studies that makes difficult the comparison of the PAG organisation between species. Therefore, to study the PAG in all mammals including human, the most relevant in vivo imaging method seems to be the magnetic resonance imaging (MRI). The purpose of this review was to summarize the knowledge of the anatomical organisation of the PAG in mammals and highlights the benefits of MRI methods to extend this knowledge. Results obtained by MRI so far support the conclusions of ex vivo studies, especially to describe the subdivisions and the connections of the PAG. In these latter, diffusion-weighted MRI and functional connectivity seem the most appropriate methods. In conclusion firstly, the MRI seems to be the best judicious method to compare species and improve the comprehension of the role of the PAG. Secondly, MRI is an in vivo method aimed to manage repeated measures in the same cohort of subjects allowing to study the impact of aging and the development on the anatomical organisation of the PAG.
Involvement of spinal release of α-neo-endorphin on the antinociceptive effect of TAPA
2013, Peptides
Citation Excerpt :
We observed that the spinal antinociception of TAPA was significantly suppressed by i.t.-pretreatment with naloxonazine or i.t.-co-administration of d-Pro2-endomorphin-2, but not i.t.-co-administration of d-Pro2-Tyr-W-MIF-1 or d-Pro2-endomorphin-1, suggesting that the spinal antinociception of TAPA is selectively mediated through the activation of μ1-opioid receptors. This result was confirmed in CXBK mice, whose μ1-opioid receptors are naturally reduced [11,12]. In CXBK mice, the spinal antinociception of TAPA was significantly suppressed and the dose–response curve of TAPA for antinociception was markedly shifted to the right compared to C57BL/6ByJ mice.
The antinociceptive effect of i.t.-administered Tyr-d-Arg-Phe-β-Ala (TAPA), an N-terminal tetrapeptide analog of dermorphin, was characterized in ddY mice. In the mouse tail-flick test, TAPA administered i.t. produced a potent antinociception. The antinociception induced by TAPA was significantly attenuated by i.t. pretreatment with the κ-opioid receptor antagonist nor-binaltorphimine, as well as by the μ-opioid receptor antagonist β-funaltrexamine and the μ₁-opioid receptor antagonist naloxonazine. TAPA-induced antinociception was also significantly suppressed by co-administration of the μ₁-opioid receptor antagonist Tyr-d-Pro-Phe-Phe-NH₂ (d-Pro²-endomorphin-2) but not by co-administration of the μ₂-opioid receptor antagonists Tyr-d-Pro-Trp-Phe-NH₂ (d-Pro²-endomorphin-1) and Tyr-d-Pro-Trp-Gly-NH₂ (d-Pro²-Tyr-W-MIF-1). In CXBK mice whose μ₁-opioid receptors were naturally reduced, the antinociceptive effect of TAPA was markedly suppressed compared to the parental strain C57BL/6ByJ mice. Moreover, the antinociception induced by TAPA was significantly attenuated by i.t. pretreatment with antiserum against the endogenous κ-opioid peptide α-neo-endorphin but not antisera against other endogenous opioid peptides. In prodynorphin-deficient mice, the antinociceptive effect of TAPA was significantly reduced compared to wild-type mice. These results suggest that the spinal antinociception induced by TAPA is mediated in part through the release of α-neo-endorphin in the spinal cord via activation of spinal μ₁-opioid receptors.
Ethanol drinking-in-the-dark facilitates behavioral sensitization to ethanol in C57BL/6J, BALB/cByJ, but not in mu-opioid receptor deficient CXBK mice
2012, Pharmacology Biochemistry and Behavior
Citation Excerpt :
The CXBK line is a member of the recombinant inbred CXB set originally produced (Bailey, 1971) from C57BL/6By and BALB/cBy progenitors. Baran et al. (1975) initially showed that CXBK mice had lower levels of brain mu-opioid receptor agonist binding, a result later found by other authors (Moskowitz and Goodman, 1985). Confirming these data, we found that both BALB and B6 mice showed higher levels of mu-opioid receptor protein in the striatum, cerebellum, hypothalamus and ventral midbrain than those obtained with CXBK brains.
Neuroplasticity associated with drug-induced behavioral sensitization has been associated with excessive drug pursuit and consumption characteristic of addiction. Repeated intraperitoneal (ip) injections of ethanol (EtOH) can induce psychomotor sensitization in mice. In terms of its clinical relevance, however, it is important to determine whether this phenomenon can also be produced by voluntary EtOH consumption.
The present investigation used a drinking-in-the-dark (DID) methodology to induce high levels of EtOH drinking in mice; EtOH replaces water for 2 or 4 h, starting 3 h after the beginning of the dark cycle. Animals followed a 3-week DID protocol prior to an evaluation of EtOH-induced locomotor activity (acute and repeated EtOH). For the first week, animals had access to 20% EtOH. On weeks 2 and 3, different concentrations of EtOH (10, 20 or 30%) were used. Three different inbred strains of mice were used: C57BL/6J (B6), BALB/cByJ (BALB), and CXBK. The CXBK mouse line was used because of its reduced expression and functioning of brain mu-opioid receptors, which have been suggested to participate in the development of EtOH-induced sensitization. B6 and BALB mice were used as controls.
B6 and CXBK mice presented comparable levels of EtOH drinking (approx. 3 g/kg in 2 h), that were higher than those showed by BALB. All animals, regardless of genotype, adjusted volume of EtOH intake to obtain stable g/kg of EtOH across concentrations. Previous EtOH DID produced (B6) or potentiated (BALB) sensitization to EtOH; this effect was not seen in CXBK. Western blot analysis showed a reduced number of mu-opioid receptors in several brain regions of CXBK as compared to that of B6 and BALB mice.
In summary, here we show that the DID methodology can be used to trigger EtOH-induced neuroplasticity supporting psychomotor sensitization, a process that might require participation of mu-opioid receptors.

View all citing articles on Scopus

View full text

Autoradiographic analysis of mu1, mu2, and delta opioid binding in the central nervous system of C57BL/6BY and CXBK (opioid receptor-deficient) mice

Abstract

Brain Research

Brain Research

Brain Research

Life Sci.

Brain Research

Neurosci. Lett.

Brain Research

Brain Research

Eur. J. Pharmacol.

Brain Research

Brain Research

Brain Research

Toxicol. Appl. Pharmacol.

Life Sci.

Eur. J. Pharmacol.

Eur. J. Pharmacol.

Multiple opiate receptors: different regional distribution in the brain and differential binding of opiates and opioid peptides

Mol. Pharmacol.

Opiate receptors: different ligand affinity in various brain regions

Different brain areas mediate the analgesic and epileptic properties of enkephalins

Science

Dynorphin-(1–13), an extraordinarily potent opioid peptide

Kappa opiate receptors localized by autoradiography to deep layers of cerebral cortex: relation to sedative effects

Differentiation of delta and mu opiate receptor localization by light microscopic autoradiography

In vitro autoradiography of opiate receptors in rat brain suggests loci of ‘opiatergic’ pathways

Is there some indication from behavioral effects of endorphin for their involvement in psychiatric diseases?

Study on morphine induced opacity in mouse lens: different age and strain

Res. Commun. Sub. Abuse

Identification of two related pentapeptides from the brain with potent opiate agonist activity

Nature (London)

Narcotic analgesics

Regional distribution of opiate receptor binding in monkey and human brain

Nature (London)

Opioid binding properties of brain and peripheral tissues: evidence for heterogeneity in opioid ligand binding sites

J. Pharmacol. Exp. Ther.

Opiate receptor gradients in monkey cerebral cortex: correspondence with sensory processing hierarchies

Science

Autoradiographic analysis of mu₁, mu₂, and delta opioid binding in the central nervous system of C57BL/6BY and CXBK (opioid receptor-deficient) mice