Cloning and pharmacological characterization of the equine adenosine A 3 receptor

C. I. Brandon, M. Vandenplas, H. Dookwah, Thomas F. Murray

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The aim of this study was to establish a heterologous expression system for the equine adenosine A3 receptor (eA3-R) in an effort to ascertain its pharmacologic profile. Initially, radioligand binding assays identified clones expressing the eA3-R in human embryonic kidney cells (HEK) based on the specific binding of [125I]AB-MECA. Subsequently, adenylate cyclase assays were utilized to demonstrate functional coupling of the eA3-R to the G-protein/adenylate cyclase system. Equilibrium competition binding assays were then performed using selective and non-selective A3 agonists and antagonists. Results from these experiments revealed a rank order of agonist potency to be IB-MECA > NECA > CGS21680, and an antagonist potency of MRS1220 > ZM241385 > 8-p-sulphophenyltheophylline; these rank orders were in agreement with that of other mammalian A3-R's. Lastly, NF-κB reporter gene assays revealed an IB-MECA concentration-dependent inhibition of TNFα-stimulated NF-κB activity. These results indicate that the heterologously expressed eA3-R is functional, has a pharmacological profile similar to that of other mammalian A3 receptors, and its activation has an inhibitory effect on a key regulatory pathway in the inflammatory response. Thus, the eA3-R may serve as a pharmacological target in the treatment of equine inflammatory disease.

Original languageEnglish
Pages (from-to)255-263
Number of pages9
JournalJournal of Veterinary Pharmacology and Therapeutics
Volume29
Issue number4
DOIs
StatePublished - Aug 2006
Externally publishedYes

Fingerprint

Adenosine A3 Receptors
adenosine
Adenosine
Horses
Organism Cloning
molecular cloning
Pharmacology
horses
receptors
adenylate cyclase
Adenylyl Cyclases
assays
agonists
Adenosine-5'-(N-ethylcarboxamide)
Horse Diseases
antagonists
Radioligand Assay
Reporter Genes
GTP-Binding Proteins
kidney cells

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • veterinary(all)

Cite this

Cloning and pharmacological characterization of the equine adenosine A 3 receptor. / Brandon, C. I.; Vandenplas, M.; Dookwah, H.; Murray, Thomas F.

In: Journal of Veterinary Pharmacology and Therapeutics, Vol. 29, No. 4, 08.2006, p. 255-263.

Research output: Contribution to journalArticle

@article{4a15a4c8e07248a8bc70563bdeef3736,
title = "Cloning and pharmacological characterization of the equine adenosine A 3 receptor",
abstract = "The aim of this study was to establish a heterologous expression system for the equine adenosine A3 receptor (eA3-R) in an effort to ascertain its pharmacologic profile. Initially, radioligand binding assays identified clones expressing the eA3-R in human embryonic kidney cells (HEK) based on the specific binding of [125I]AB-MECA. Subsequently, adenylate cyclase assays were utilized to demonstrate functional coupling of the eA3-R to the G-protein/adenylate cyclase system. Equilibrium competition binding assays were then performed using selective and non-selective A3 agonists and antagonists. Results from these experiments revealed a rank order of agonist potency to be IB-MECA > NECA > CGS21680, and an antagonist potency of MRS1220 > ZM241385 > 8-p-sulphophenyltheophylline; these rank orders were in agreement with that of other mammalian A3-R's. Lastly, NF-κB reporter gene assays revealed an IB-MECA concentration-dependent inhibition of TNFα-stimulated NF-κB activity. These results indicate that the heterologously expressed eA3-R is functional, has a pharmacological profile similar to that of other mammalian A3 receptors, and its activation has an inhibitory effect on a key regulatory pathway in the inflammatory response. Thus, the eA3-R may serve as a pharmacological target in the treatment of equine inflammatory disease.",
author = "Brandon, {C. I.} and M. Vandenplas and H. Dookwah and Murray, {Thomas F.}",
year = "2006",
month = "8",
doi = "10.1111/j.1365-2885.2006.00748.x",
language = "English",
volume = "29",
pages = "255--263",
journal = "Journal of Veterinary Pharmacology and Therapeutics",
issn = "0140-7783",
publisher = "Wiley-Blackwell",
number = "4",

}

TY - JOUR

T1 - Cloning and pharmacological characterization of the equine adenosine A 3 receptor

AU - Brandon, C. I.

AU - Vandenplas, M.

AU - Dookwah, H.

AU - Murray, Thomas F.

PY - 2006/8

Y1 - 2006/8

N2 - The aim of this study was to establish a heterologous expression system for the equine adenosine A3 receptor (eA3-R) in an effort to ascertain its pharmacologic profile. Initially, radioligand binding assays identified clones expressing the eA3-R in human embryonic kidney cells (HEK) based on the specific binding of [125I]AB-MECA. Subsequently, adenylate cyclase assays were utilized to demonstrate functional coupling of the eA3-R to the G-protein/adenylate cyclase system. Equilibrium competition binding assays were then performed using selective and non-selective A3 agonists and antagonists. Results from these experiments revealed a rank order of agonist potency to be IB-MECA > NECA > CGS21680, and an antagonist potency of MRS1220 > ZM241385 > 8-p-sulphophenyltheophylline; these rank orders were in agreement with that of other mammalian A3-R's. Lastly, NF-κB reporter gene assays revealed an IB-MECA concentration-dependent inhibition of TNFα-stimulated NF-κB activity. These results indicate that the heterologously expressed eA3-R is functional, has a pharmacological profile similar to that of other mammalian A3 receptors, and its activation has an inhibitory effect on a key regulatory pathway in the inflammatory response. Thus, the eA3-R may serve as a pharmacological target in the treatment of equine inflammatory disease.

AB - The aim of this study was to establish a heterologous expression system for the equine adenosine A3 receptor (eA3-R) in an effort to ascertain its pharmacologic profile. Initially, radioligand binding assays identified clones expressing the eA3-R in human embryonic kidney cells (HEK) based on the specific binding of [125I]AB-MECA. Subsequently, adenylate cyclase assays were utilized to demonstrate functional coupling of the eA3-R to the G-protein/adenylate cyclase system. Equilibrium competition binding assays were then performed using selective and non-selective A3 agonists and antagonists. Results from these experiments revealed a rank order of agonist potency to be IB-MECA > NECA > CGS21680, and an antagonist potency of MRS1220 > ZM241385 > 8-p-sulphophenyltheophylline; these rank orders were in agreement with that of other mammalian A3-R's. Lastly, NF-κB reporter gene assays revealed an IB-MECA concentration-dependent inhibition of TNFα-stimulated NF-κB activity. These results indicate that the heterologously expressed eA3-R is functional, has a pharmacological profile similar to that of other mammalian A3 receptors, and its activation has an inhibitory effect on a key regulatory pathway in the inflammatory response. Thus, the eA3-R may serve as a pharmacological target in the treatment of equine inflammatory disease.

UR - http://www.scopus.com/inward/record.url?scp=33745598818&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745598818&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2885.2006.00748.x

DO - 10.1111/j.1365-2885.2006.00748.x

M3 - Article

VL - 29

SP - 255

EP - 263

JO - Journal of Veterinary Pharmacology and Therapeutics

JF - Journal of Veterinary Pharmacology and Therapeutics

SN - 0140-7783

IS - 4

ER -