Converging protein kinase pathways mediate adenylyl cyclase superactivation upon chronic δ-opioid agonist treatment

Eva V. Varga, Marc K. Rubenzik, Dagmar Stropova, Masano Sugiyama, Vanessa Grife, Victor J Hruby, Kenner C. Rice, William R Roeske, Henry I. Yamamura

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Adenylyl cyclase (AC) superactivation is thought to play an important role in opioid tolerance, dependence, and withdrawal. In the present study, we investigated the involvement of protein kinases in chronic δ-opioid agonist-mediated AC superactivation in Chinese hamster ovary (CHO) cells stably expressing the human δ-opioid receptor (hDOR/CHO). Maximal forskolin-stimulated cAMP formation in hDOR/CHO cells increased by 472 ± 91, 399 ± 2, and 433 ± 73% after chronic treatment with the δ-opioid agonists (+)-4-[(αR)-α-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3- methoxy-benzyl]-N,N-diethyl benzamide (SNC 80), [D-Pen2,D-Pen5]-enkephalin, and deltorphin II, respectively. Concurrently, chronic SNC 80 (1 μM, 4-h) treatment augmented 32p incorporation into a 200-kDa protein immunoreactive with the ACV/VI antibody by 300 ± 60% in hDOR/CHO cell lysates. The calmodulin antagonist calmidazolium significantly attenuated chronic deltorphin II-mediated AC superactivation. Tyrosine kinase (genistein) and protein kinase C (chelerythrine) inhibitors individually had minimal effect on chronic δ-opioid agonist-mediated AC superactivation. Conversely, simultaneous treatment with both genistein and chelerythrine significantly attenuated AC superactivation. Because we showed previously that the Raf-1 inhibitor 3-(3,5-dibromo-4-hydroxybenzylidene-5-iodo-1,3-dihydro-indol-2-one (GW5074) attenuates AC superactivation, we hypothesize that parallel calmidazolium-, chelerythrine-, and genistein-sensitive pathways converge at Raf-1 to mediate AC superactivation by phosphorylating AC VI in hDOR/CHO cells.

Original languageEnglish (US)
Pages (from-to)109-115
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume306
Issue number1
DOIs
StatePublished - Jul 1 2003

Fingerprint

Adenylyl Cyclases
Protein Kinases
Opioid Analgesics
Cricetulus
Ovary
calmidazolium
Genistein
D-Penicillamine (2,5)-Enkephalin
Opioid Receptors
Colforsin
Calmodulin
Protein-Tyrosine Kinases
Protein Kinase C
Antibodies
chelerythrine
Proteins

ASJC Scopus subject areas

  • Pharmacology

Cite this

Converging protein kinase pathways mediate adenylyl cyclase superactivation upon chronic δ-opioid agonist treatment. / Varga, Eva V.; Rubenzik, Marc K.; Stropova, Dagmar; Sugiyama, Masano; Grife, Vanessa; Hruby, Victor J; Rice, Kenner C.; Roeske, William R; Yamamura, Henry I.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 306, No. 1, 01.07.2003, p. 109-115.

Research output: Contribution to journalArticle

Varga, Eva V. ; Rubenzik, Marc K. ; Stropova, Dagmar ; Sugiyama, Masano ; Grife, Vanessa ; Hruby, Victor J ; Rice, Kenner C. ; Roeske, William R ; Yamamura, Henry I. / Converging protein kinase pathways mediate adenylyl cyclase superactivation upon chronic δ-opioid agonist treatment. In: Journal of Pharmacology and Experimental Therapeutics. 2003 ; Vol. 306, No. 1. pp. 109-115.
@article{0bbeb88c70ab49a981eb959e20fb88be,
title = "Converging protein kinase pathways mediate adenylyl cyclase superactivation upon chronic δ-opioid agonist treatment",
abstract = "Adenylyl cyclase (AC) superactivation is thought to play an important role in opioid tolerance, dependence, and withdrawal. In the present study, we investigated the involvement of protein kinases in chronic δ-opioid agonist-mediated AC superactivation in Chinese hamster ovary (CHO) cells stably expressing the human δ-opioid receptor (hDOR/CHO). Maximal forskolin-stimulated cAMP formation in hDOR/CHO cells increased by 472 ± 91, 399 ± 2, and 433 ± 73{\%} after chronic treatment with the δ-opioid agonists (+)-4-[(αR)-α-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3- methoxy-benzyl]-N,N-diethyl benzamide (SNC 80), [D-Pen2,D-Pen5]-enkephalin, and deltorphin II, respectively. Concurrently, chronic SNC 80 (1 μM, 4-h) treatment augmented 32p incorporation into a 200-kDa protein immunoreactive with the ACV/VI antibody by 300 ± 60{\%} in hDOR/CHO cell lysates. The calmodulin antagonist calmidazolium significantly attenuated chronic deltorphin II-mediated AC superactivation. Tyrosine kinase (genistein) and protein kinase C (chelerythrine) inhibitors individually had minimal effect on chronic δ-opioid agonist-mediated AC superactivation. Conversely, simultaneous treatment with both genistein and chelerythrine significantly attenuated AC superactivation. Because we showed previously that the Raf-1 inhibitor 3-(3,5-dibromo-4-hydroxybenzylidene-5-iodo-1,3-dihydro-indol-2-one (GW5074) attenuates AC superactivation, we hypothesize that parallel calmidazolium-, chelerythrine-, and genistein-sensitive pathways converge at Raf-1 to mediate AC superactivation by phosphorylating AC VI in hDOR/CHO cells.",
author = "Varga, {Eva V.} and Rubenzik, {Marc K.} and Dagmar Stropova and Masano Sugiyama and Vanessa Grife and Hruby, {Victor J} and Rice, {Kenner C.} and Roeske, {William R} and Yamamura, {Henry I.}",
year = "2003",
month = "7",
day = "1",
doi = "10.1124/jpet.103.049643",
language = "English (US)",
volume = "306",
pages = "109--115",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "1",

}

TY - JOUR

T1 - Converging protein kinase pathways mediate adenylyl cyclase superactivation upon chronic δ-opioid agonist treatment

AU - Varga, Eva V.

AU - Rubenzik, Marc K.

AU - Stropova, Dagmar

AU - Sugiyama, Masano

AU - Grife, Vanessa

AU - Hruby, Victor J

AU - Rice, Kenner C.

AU - Roeske, William R

AU - Yamamura, Henry I.

PY - 2003/7/1

Y1 - 2003/7/1

N2 - Adenylyl cyclase (AC) superactivation is thought to play an important role in opioid tolerance, dependence, and withdrawal. In the present study, we investigated the involvement of protein kinases in chronic δ-opioid agonist-mediated AC superactivation in Chinese hamster ovary (CHO) cells stably expressing the human δ-opioid receptor (hDOR/CHO). Maximal forskolin-stimulated cAMP formation in hDOR/CHO cells increased by 472 ± 91, 399 ± 2, and 433 ± 73% after chronic treatment with the δ-opioid agonists (+)-4-[(αR)-α-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3- methoxy-benzyl]-N,N-diethyl benzamide (SNC 80), [D-Pen2,D-Pen5]-enkephalin, and deltorphin II, respectively. Concurrently, chronic SNC 80 (1 μM, 4-h) treatment augmented 32p incorporation into a 200-kDa protein immunoreactive with the ACV/VI antibody by 300 ± 60% in hDOR/CHO cell lysates. The calmodulin antagonist calmidazolium significantly attenuated chronic deltorphin II-mediated AC superactivation. Tyrosine kinase (genistein) and protein kinase C (chelerythrine) inhibitors individually had minimal effect on chronic δ-opioid agonist-mediated AC superactivation. Conversely, simultaneous treatment with both genistein and chelerythrine significantly attenuated AC superactivation. Because we showed previously that the Raf-1 inhibitor 3-(3,5-dibromo-4-hydroxybenzylidene-5-iodo-1,3-dihydro-indol-2-one (GW5074) attenuates AC superactivation, we hypothesize that parallel calmidazolium-, chelerythrine-, and genistein-sensitive pathways converge at Raf-1 to mediate AC superactivation by phosphorylating AC VI in hDOR/CHO cells.

AB - Adenylyl cyclase (AC) superactivation is thought to play an important role in opioid tolerance, dependence, and withdrawal. In the present study, we investigated the involvement of protein kinases in chronic δ-opioid agonist-mediated AC superactivation in Chinese hamster ovary (CHO) cells stably expressing the human δ-opioid receptor (hDOR/CHO). Maximal forskolin-stimulated cAMP formation in hDOR/CHO cells increased by 472 ± 91, 399 ± 2, and 433 ± 73% after chronic treatment with the δ-opioid agonists (+)-4-[(αR)-α-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3- methoxy-benzyl]-N,N-diethyl benzamide (SNC 80), [D-Pen2,D-Pen5]-enkephalin, and deltorphin II, respectively. Concurrently, chronic SNC 80 (1 μM, 4-h) treatment augmented 32p incorporation into a 200-kDa protein immunoreactive with the ACV/VI antibody by 300 ± 60% in hDOR/CHO cell lysates. The calmodulin antagonist calmidazolium significantly attenuated chronic deltorphin II-mediated AC superactivation. Tyrosine kinase (genistein) and protein kinase C (chelerythrine) inhibitors individually had minimal effect on chronic δ-opioid agonist-mediated AC superactivation. Conversely, simultaneous treatment with both genistein and chelerythrine significantly attenuated AC superactivation. Because we showed previously that the Raf-1 inhibitor 3-(3,5-dibromo-4-hydroxybenzylidene-5-iodo-1,3-dihydro-indol-2-one (GW5074) attenuates AC superactivation, we hypothesize that parallel calmidazolium-, chelerythrine-, and genistein-sensitive pathways converge at Raf-1 to mediate AC superactivation by phosphorylating AC VI in hDOR/CHO cells.

UR - http://www.scopus.com/inward/record.url?scp=0038340971&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038340971&partnerID=8YFLogxK

U2 - 10.1124/jpet.103.049643

DO - 10.1124/jpet.103.049643

M3 - Article

C2 - 12660310

AN - SCOPUS:0038340971

VL - 306

SP - 109

EP - 115

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 1

ER -