Knock-In Mice with NOP-eGFP Receptors Identify Receptor Cellular and Regional Localization
Akihiko Ozawa,Gloria Brunori,Daniela Mercatelli,Jinhua Wu,Andrea Cippitelli,Bende Zou,Xinmin Simon Xie,Melissa Williams,Nurulain T. Zaveri,Sarah A. Low,Grégory Scherrer,Brigitte L. Kieffer,Lawrence Toll +12 more
TLDR
The NOP-eGFP receptor was present in brain of homozygous knock-in animals in concentrations somewhat higher than in wild-type mice and was functional when tested for stimulation of GTPγS binding in vitro and in patch-clamp electrophysiology in dorsal root ganglia (DRG) neurons and hippocampal slices as discussed by the authors.Abstract:
The nociceptin/orphanin FQ (NOP) receptor, the fourth member of the opioid receptor family, is involved in many processes common to the opioid receptors including pain and drug abuse. To better characterize receptor location and trafficking, knock-in mice were created by inserting the gene encoding enhanced green fluorescent protein (eGFP) into the NOP receptor gene ( Oprl1 ) and producing mice expressing a functional NOP-eGFP C-terminal fusion in place of the native NOP receptor. The NOP-eGFP receptor was present in brain of homozygous knock-in animals in concentrations somewhat higher than in wild-type mice and was functional when tested for stimulation of [ 35 S]GTPγS binding in vitro and in patch-clamp electrophysiology in dorsal root ganglia (DRG) neurons and hippocampal slices. Inhibition of morphine analgesia was equivalent when tested in knock-in and wild-type mice. Imaging revealed detailed neuroanatomy in brain, spinal cord, and DRG and was generally consistent with in vitro autoradiographic imaging of receptor location. Multicolor immunohistochemistry identified cells coexpressing various spinal cord and DRG cellular markers, as well as coexpression with μ-opioid receptors in DRG and brain regions. Both in tissue slices and primary cultures, the NOP-eGFP receptors appear throughout the cell body and in processes. These knock-in mice have NOP receptors that function both in vitro and in vivo and appear to be an exceptional tool to study receptor neuroanatomy and correlate with NOP receptor function. SIGNIFICANCE STATEMENT The NOP receptor, the fourth member of the opioid receptor family, is involved in pain, drug abuse, and a number of other CNS processes. The regional and cellular distribution has been difficult to determine due to lack of validated antibodies for immunohistochemical analysis. To provide a new tool for the investigation of receptor localization, we have produced knock-in mice with a fluorescent-tagged NOP receptor in place of the native NOP receptor. These knock-in mice have NOP receptors that function both in vitro and in vivo and have provided a detailed characterization of NOP receptors in brain, spinal cord, and DRG neurons. They appear to be an exceptional tool to study receptor neuroanatomy and correlate with NOP receptor function.read more
Citations
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Journal ArticleDOI
Endogenous and Exogenous Opioids in Pain.
TL;DR: This review describes the functional organization and pharmacology of the endogenous opioid system and discusses the loci of opioid analgesic action along peripheral and central pain pathways, emphasizing the pain-relieving properties of opioids against the affective dimension of the pain experience.
Journal ArticleDOI
Nociceptin/Orphanin FQ Receptor Structure, Signaling, Ligands, Functions, and Interactions with Opioid Systems
TL;DR: How NOP pharmacology intersects, contrasts, and interacts with the mu opioid receptor in terms of tertiary structure and mechanism of receptor activation; location of receptors in the central nervous system; mechanisms of desensitization and downregulation; cellular actions; intracellular signal transduction pathways; and behavioral actions with respect to analgesia, tolerance, dependence, and reward is discussed.
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Targeting multiple opioid receptors - improved analgesics with reduced side effects?
Thomas Günther,Pooja Dasgupta,Anika Mann,Elke Miess,Andrea Kliewer,Sebastian Fritzwanker,Ralph Steinborn,Stefan Schulz +7 more
TL;DR: The main focus of this review is to assess the paradigm of opioid ligands targeting multiple receptors with a single chemical entity and reflect on this rationale by discussing the biological actions of particular multi‐opioid receptor ligands, but not on their medicinal chemistry and design.
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A Paranigral VTA Nociceptin Circuit that Constrains Motivation for Reward.
Kyle E. Parker,Christian E. Pedersen,Adrian M. Gomez,Skylar M. Spangler,Marie C. Walicki,Shelley Y. Feng,Sarah Stewart,James M. Otis,Ream Al-Hasani,Jordan G. McCall,Kristina Sakers,Dionnet L. Bhatti,Bryan A. Copits,Robert W. Gereau,Thomas C. Jhou,Thomas J. Kash,Joseph D. Dougherty,Garret D. Stuber,Michael R. Bruchas +18 more
TL;DR: Insight is provided into neuromodulatory circuits that regulate motivated behaviors through identification of a previously unknown neuropeptide-containing pnVTA nucleus that limits motivation for rewards.
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New Technologies for Elucidating Opioid Receptor Function
Michael R. Bruchas,Bryan L. Roth +1 more
TL;DR: How newly engineered opioid receptor-based chemogenetic and optogenetic tools, and new mouse lines, are expanding and transforming the understanding of opioid function and, perhaps, paving the way for new therapeutics is described.
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