Ligand-directed signaling at the beta3-adrenoceptor produced by 3-(2-ethylphenoxy)-1-[(1,S)-1,2,3,4-tetrahydronapth-1-ylamino]-2S-2-propanol oxalate (SR59230A) relative to receptor agonists

Masaaki Sato, Takahiro Horinouchi, Dana Sabine Hutchinson, Bronwyn Anne Evans, Roger James Summers

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Abstract

This study examines signaling pathways activated by the mouse beta3-adrenoceptor (AR) expressed in Chinese hamster ovary cells at high (CHObeta3H) or low (CHObeta3L) levels. Functional responses included extracellular acidification rate (ECAR), cyclic AMP accumulation, and p38 MAPK or Erk1/2 phosphorylation. (-)-Isoproterenol and the beta3-AR agonist CL316243 caused concentration-dependent increases in cAMP accumulation and ECAR in CHObeta3H and CHObeta3L cells. For cAMP accumulation, the beta3-AR ligand SR59230A was a partial agonist in CHObeta3H and an antagonist in CHObeta3L cells but for ECAR was an agonist at both expression levels. This suggested that SR59230A which is normally regarded as an antagonist can selectively activate pathways leading to ECAR. Examination of the pathways stimulated by (-)-isoproterenol, CL316243 and SR59230A for both ECAR and cAMP accumulation suggested that the cAMP pathway predominates in CHObeta3H cells while p38 MAPK is a major contributor to ECAR in CHObeta3L cells and was the sole contributor to responses to SR59230A. Western blots of p38 MAPK and Erk1/2 phosphorylation confirmed that MAPKs are activated in CHObeta3H and CHObeta3L cells by CL316243 and SR59230A but that SR59230A has much higher efficacy. In addition, p38 MAPK phosphorylation displayed differences in drug potency and efficacy between CHObeta3H and CHObeta3L cells related to inhibition of the response by cAMP. Thus CL316243 and SR59230A display reversed orders of efficacy for cAMP accumulation compared to Erk1/2 and p38 MAPK phosphorylation providing a strong indication of ligand-directed signaling.
Original languageEnglish
Pages (from-to)1359 - 1368
Number of pages10
JournalMolecular Pharmacology
Volume72
Issue number5
DOIs
Publication statusPublished - 2007

Cite this

@article{244138b0560e48749365fcdcaea1fd4e,
title = "Ligand-directed signaling at the beta3-adrenoceptor produced by 3-(2-ethylphenoxy)-1-[(1,S)-1,2,3,4-tetrahydronapth-1-ylamino]-2S-2-propanol oxalate (SR59230A) relative to receptor agonists",
abstract = "This study examines signaling pathways activated by the mouse beta3-adrenoceptor (AR) expressed in Chinese hamster ovary cells at high (CHObeta3H) or low (CHObeta3L) levels. Functional responses included extracellular acidification rate (ECAR), cyclic AMP accumulation, and p38 MAPK or Erk1/2 phosphorylation. (-)-Isoproterenol and the beta3-AR agonist CL316243 caused concentration-dependent increases in cAMP accumulation and ECAR in CHObeta3H and CHObeta3L cells. For cAMP accumulation, the beta3-AR ligand SR59230A was a partial agonist in CHObeta3H and an antagonist in CHObeta3L cells but for ECAR was an agonist at both expression levels. This suggested that SR59230A which is normally regarded as an antagonist can selectively activate pathways leading to ECAR. Examination of the pathways stimulated by (-)-isoproterenol, CL316243 and SR59230A for both ECAR and cAMP accumulation suggested that the cAMP pathway predominates in CHObeta3H cells while p38 MAPK is a major contributor to ECAR in CHObeta3L cells and was the sole contributor to responses to SR59230A. Western blots of p38 MAPK and Erk1/2 phosphorylation confirmed that MAPKs are activated in CHObeta3H and CHObeta3L cells by CL316243 and SR59230A but that SR59230A has much higher efficacy. In addition, p38 MAPK phosphorylation displayed differences in drug potency and efficacy between CHObeta3H and CHObeta3L cells related to inhibition of the response by cAMP. Thus CL316243 and SR59230A display reversed orders of efficacy for cAMP accumulation compared to Erk1/2 and p38 MAPK phosphorylation providing a strong indication of ligand-directed signaling.",
author = "Masaaki Sato and Takahiro Horinouchi and Hutchinson, {Dana Sabine} and Evans, {Bronwyn Anne} and Summers, {Roger James}",
year = "2007",
doi = "10.1124/mol.107.03533",
language = "English",
volume = "72",
pages = "1359 -- 1368",
journal = "Molecular Pharmacology",
issn = "1521-0111",
publisher = "ACS Books",
number = "5",

}

TY - JOUR

T1 - Ligand-directed signaling at the beta3-adrenoceptor produced by 3-(2-ethylphenoxy)-1-[(1,S)-1,2,3,4-tetrahydronapth-1-ylamino]-2S-2-propanol oxalate (SR59230A) relative to receptor agonists

AU - Sato, Masaaki

AU - Horinouchi, Takahiro

AU - Hutchinson, Dana Sabine

AU - Evans, Bronwyn Anne

AU - Summers, Roger James

PY - 2007

Y1 - 2007

N2 - This study examines signaling pathways activated by the mouse beta3-adrenoceptor (AR) expressed in Chinese hamster ovary cells at high (CHObeta3H) or low (CHObeta3L) levels. Functional responses included extracellular acidification rate (ECAR), cyclic AMP accumulation, and p38 MAPK or Erk1/2 phosphorylation. (-)-Isoproterenol and the beta3-AR agonist CL316243 caused concentration-dependent increases in cAMP accumulation and ECAR in CHObeta3H and CHObeta3L cells. For cAMP accumulation, the beta3-AR ligand SR59230A was a partial agonist in CHObeta3H and an antagonist in CHObeta3L cells but for ECAR was an agonist at both expression levels. This suggested that SR59230A which is normally regarded as an antagonist can selectively activate pathways leading to ECAR. Examination of the pathways stimulated by (-)-isoproterenol, CL316243 and SR59230A for both ECAR and cAMP accumulation suggested that the cAMP pathway predominates in CHObeta3H cells while p38 MAPK is a major contributor to ECAR in CHObeta3L cells and was the sole contributor to responses to SR59230A. Western blots of p38 MAPK and Erk1/2 phosphorylation confirmed that MAPKs are activated in CHObeta3H and CHObeta3L cells by CL316243 and SR59230A but that SR59230A has much higher efficacy. In addition, p38 MAPK phosphorylation displayed differences in drug potency and efficacy between CHObeta3H and CHObeta3L cells related to inhibition of the response by cAMP. Thus CL316243 and SR59230A display reversed orders of efficacy for cAMP accumulation compared to Erk1/2 and p38 MAPK phosphorylation providing a strong indication of ligand-directed signaling.

AB - This study examines signaling pathways activated by the mouse beta3-adrenoceptor (AR) expressed in Chinese hamster ovary cells at high (CHObeta3H) or low (CHObeta3L) levels. Functional responses included extracellular acidification rate (ECAR), cyclic AMP accumulation, and p38 MAPK or Erk1/2 phosphorylation. (-)-Isoproterenol and the beta3-AR agonist CL316243 caused concentration-dependent increases in cAMP accumulation and ECAR in CHObeta3H and CHObeta3L cells. For cAMP accumulation, the beta3-AR ligand SR59230A was a partial agonist in CHObeta3H and an antagonist in CHObeta3L cells but for ECAR was an agonist at both expression levels. This suggested that SR59230A which is normally regarded as an antagonist can selectively activate pathways leading to ECAR. Examination of the pathways stimulated by (-)-isoproterenol, CL316243 and SR59230A for both ECAR and cAMP accumulation suggested that the cAMP pathway predominates in CHObeta3H cells while p38 MAPK is a major contributor to ECAR in CHObeta3L cells and was the sole contributor to responses to SR59230A. Western blots of p38 MAPK and Erk1/2 phosphorylation confirmed that MAPKs are activated in CHObeta3H and CHObeta3L cells by CL316243 and SR59230A but that SR59230A has much higher efficacy. In addition, p38 MAPK phosphorylation displayed differences in drug potency and efficacy between CHObeta3H and CHObeta3L cells related to inhibition of the response by cAMP. Thus CL316243 and SR59230A display reversed orders of efficacy for cAMP accumulation compared to Erk1/2 and p38 MAPK phosphorylation providing a strong indication of ligand-directed signaling.

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U2 - 10.1124/mol.107.03533

DO - 10.1124/mol.107.03533

M3 - Article

VL - 72

SP - 1359

EP - 1368

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 1521-0111

IS - 5

ER -