Functional analysis of cloned opioid receptors in transfected cell lines

E T Piros, T G Hales, C J Evans

Research output: Contribution to journalReview article

38 Citations (Scopus)

Abstract

Opioids modulate numerous central and peripheral processes including pain perception neuroendocrine secretion and the immune response. The opioid signal is transduced from receptors through G proteins to various different effectors. Heterogeneity exists at all levels of the transduction process. There are numerous endogenous ligands with differing selectivities for at least three distinct opioid receptors (mu, delta, kappa). G proteins activated by opioid receptors are generally of the pertussis toxin-sensitive Gi/Go class, but there are also opioid actions that are thought to involve Gq and cholera toxin-sensitive G proteins. To further complicate the issue, the actions of opioid receptors may be mediated by G-protein alpha subunits and/or beta gamma subunits. Subsequent to G protein activation several effectors are known to orchestrate the opioid signal. For example activation of opioid receptors increases phosphatidyl inositol turnover, activates K+ channels and reduces adenylyl cyclase and Ca2+ channel activities. Each of these effectors shows considerable heterogeneity. In this review we examine the opioid signal transduction mechanism. Several important questions arise: Why do opioid ligands with similar binding affinities have different potencies in functional assays? To which Ca2+ channel subtypes do opioid receptors couple? Do opioid receptors couple to Ca2+ channels through direct G protein interactions? Does the opioid-induced inhibition of vesicular release occur through modulation of multiple effectors? We are attempting to answer these questions by expressing cloned opioid receptors in GH3 cells. Using this well characterized system we can study the entire opioid signal transduction process from ligand-receptor interaction to G protein-effector coupling and subsequent inhibition of vesicular release.

Original languageEnglish
Pages (from-to)1277-85
Number of pages9
JournalNeurochemical Research
Volume21
Issue number11
DOIs
Publication statusPublished - Nov 1996

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Functional analysis
Opioid Receptors
Opioid Analgesics
Cells
GTP-Binding Proteins
Cell Line
Signal transduction
Ligands
Signal Transduction
Chemical activation
GTP-Binding Protein alpha Subunits
delta Opioid Receptor
Pain Perception
mu Opioid Receptor
Cholera Toxin
Pertussis Toxin
Phosphatidylinositols
Adenylyl Cyclases
Assays
Modulation

Keywords

  • Animals
  • Cell Line
  • Cyclic AMP/metabolism
  • GTP-Binding Proteins/metabolism
  • Humans
  • Inositol Phosphates/metabolism
  • Models, Biological
  • Narcotics/pharmacology
  • Receptors, Opioid/biosynthesis
  • Receptors, Opioid, delta/biosynthesis
  • Receptors, Opioid, kappa/biosynthesis
  • Receptors, Opioid, mu/biosynthesis
  • Recombinant Proteins/biosynthesis
  • Signal Transduction
  • Transfection/methods

Cite this

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title = "Functional analysis of cloned opioid receptors in transfected cell lines",
abstract = "Opioids modulate numerous central and peripheral processes including pain perception neuroendocrine secretion and the immune response. The opioid signal is transduced from receptors through G proteins to various different effectors. Heterogeneity exists at all levels of the transduction process. There are numerous endogenous ligands with differing selectivities for at least three distinct opioid receptors (mu, delta, kappa). G proteins activated by opioid receptors are generally of the pertussis toxin-sensitive Gi/Go class, but there are also opioid actions that are thought to involve Gq and cholera toxin-sensitive G proteins. To further complicate the issue, the actions of opioid receptors may be mediated by G-protein alpha subunits and/or beta gamma subunits. Subsequent to G protein activation several effectors are known to orchestrate the opioid signal. For example activation of opioid receptors increases phosphatidyl inositol turnover, activates K+ channels and reduces adenylyl cyclase and Ca2+ channel activities. Each of these effectors shows considerable heterogeneity. In this review we examine the opioid signal transduction mechanism. Several important questions arise: Why do opioid ligands with similar binding affinities have different potencies in functional assays? To which Ca2+ channel subtypes do opioid receptors couple? Do opioid receptors couple to Ca2+ channels through direct G protein interactions? Does the opioid-induced inhibition of vesicular release occur through modulation of multiple effectors? We are attempting to answer these questions by expressing cloned opioid receptors in GH3 cells. Using this well characterized system we can study the entire opioid signal transduction process from ligand-receptor interaction to G protein-effector coupling and subsequent inhibition of vesicular release.",
keywords = "Animals, Cell Line, Cyclic AMP/metabolism, GTP-Binding Proteins/metabolism, Humans, Inositol Phosphates/metabolism, Models, Biological, Narcotics/pharmacology, Receptors, Opioid/biosynthesis, Receptors, Opioid, delta/biosynthesis, Receptors, Opioid, kappa/biosynthesis, Receptors, Opioid, mu/biosynthesis, Recombinant Proteins/biosynthesis, Signal Transduction, Transfection/methods",
author = "Piros, {E T} and Hales, {T G} and Evans, {C J}",
year = "1996",
month = "11",
doi = "10.1007/BF02532368",
language = "English",
volume = "21",
pages = "1277--85",
journal = "Neurochemical Research",
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}

Functional analysis of cloned opioid receptors in transfected cell lines. / Piros, E T; Hales, T G; Evans, C J.

In: Neurochemical Research, Vol. 21, No. 11, 11.1996, p. 1277-85.

Research output: Contribution to journalReview article

TY - JOUR

T1 - Functional analysis of cloned opioid receptors in transfected cell lines

AU - Piros, E T

AU - Hales, T G

AU - Evans, C J

PY - 1996/11

Y1 - 1996/11

N2 - Opioids modulate numerous central and peripheral processes including pain perception neuroendocrine secretion and the immune response. The opioid signal is transduced from receptors through G proteins to various different effectors. Heterogeneity exists at all levels of the transduction process. There are numerous endogenous ligands with differing selectivities for at least three distinct opioid receptors (mu, delta, kappa). G proteins activated by opioid receptors are generally of the pertussis toxin-sensitive Gi/Go class, but there are also opioid actions that are thought to involve Gq and cholera toxin-sensitive G proteins. To further complicate the issue, the actions of opioid receptors may be mediated by G-protein alpha subunits and/or beta gamma subunits. Subsequent to G protein activation several effectors are known to orchestrate the opioid signal. For example activation of opioid receptors increases phosphatidyl inositol turnover, activates K+ channels and reduces adenylyl cyclase and Ca2+ channel activities. Each of these effectors shows considerable heterogeneity. In this review we examine the opioid signal transduction mechanism. Several important questions arise: Why do opioid ligands with similar binding affinities have different potencies in functional assays? To which Ca2+ channel subtypes do opioid receptors couple? Do opioid receptors couple to Ca2+ channels through direct G protein interactions? Does the opioid-induced inhibition of vesicular release occur through modulation of multiple effectors? We are attempting to answer these questions by expressing cloned opioid receptors in GH3 cells. Using this well characterized system we can study the entire opioid signal transduction process from ligand-receptor interaction to G protein-effector coupling and subsequent inhibition of vesicular release.

AB - Opioids modulate numerous central and peripheral processes including pain perception neuroendocrine secretion and the immune response. The opioid signal is transduced from receptors through G proteins to various different effectors. Heterogeneity exists at all levels of the transduction process. There are numerous endogenous ligands with differing selectivities for at least three distinct opioid receptors (mu, delta, kappa). G proteins activated by opioid receptors are generally of the pertussis toxin-sensitive Gi/Go class, but there are also opioid actions that are thought to involve Gq and cholera toxin-sensitive G proteins. To further complicate the issue, the actions of opioid receptors may be mediated by G-protein alpha subunits and/or beta gamma subunits. Subsequent to G protein activation several effectors are known to orchestrate the opioid signal. For example activation of opioid receptors increases phosphatidyl inositol turnover, activates K+ channels and reduces adenylyl cyclase and Ca2+ channel activities. Each of these effectors shows considerable heterogeneity. In this review we examine the opioid signal transduction mechanism. Several important questions arise: Why do opioid ligands with similar binding affinities have different potencies in functional assays? To which Ca2+ channel subtypes do opioid receptors couple? Do opioid receptors couple to Ca2+ channels through direct G protein interactions? Does the opioid-induced inhibition of vesicular release occur through modulation of multiple effectors? We are attempting to answer these questions by expressing cloned opioid receptors in GH3 cells. Using this well characterized system we can study the entire opioid signal transduction process from ligand-receptor interaction to G protein-effector coupling and subsequent inhibition of vesicular release.

KW - Animals

KW - Cell Line

KW - Cyclic AMP/metabolism

KW - GTP-Binding Proteins/metabolism

KW - Humans

KW - Inositol Phosphates/metabolism

KW - Models, Biological

KW - Narcotics/pharmacology

KW - Receptors, Opioid/biosynthesis

KW - Receptors, Opioid, delta/biosynthesis

KW - Receptors, Opioid, kappa/biosynthesis

KW - Receptors, Opioid, mu/biosynthesis

KW - Recombinant Proteins/biosynthesis

KW - Signal Transduction

KW - Transfection/methods

U2 - 10.1007/BF02532368

DO - 10.1007/BF02532368

M3 - Review article

VL - 21

SP - 1277

EP - 1285

JO - Neurochemical Research

JF - Neurochemical Research

SN - 0364-3190

IS - 11

ER -