Delta opioid receptors expressed by stably transfected jurkat cells signal through the map kinase pathway in a ras-independent manner

Nahid A. Shahabi, Yehia Daaka, Kathy McAllen, Burt Sharp

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Delta opioid receptors (DOR) are G-protein coupled 7-transmembrane receptors (GPCR), expressed by thymic and splenic T cells, that modulate interleukin (IL)-2 production and proliferation in response to concanavalin A or crosslinking the TCR. Mitogen-activated protein kinases (MAPKs) are involved in mediating intracellular responses to TCR crosslinking. In addition, MAPKs can be activated by signaling cascades that are initiated by the release of G-proteins from GPCRs. To determine whether DORs expressed by T cells signal through the MAPKs, extracellular-regulated kinases (ERKs) 1 and 2, two delta opioid peptides, deltorphin and [D-Ala2, D-Leu5]- enkephalin (DADLE), were studied in Jurkat cells that had been stably transfected with DOR (DOR-Ju.1). These peptides rapidly and dose-dependently induced ERK phosphorylation; pretreatment with naltrindole (NTI), a selective DOR antagonist: abolished this. Pertussis toxin (PTX) also inhibited phosphorylation, indicating the involvement of the G(i/o) proteins. Herbimycin A, a protein tyrosine kinase (PTK) inhibitor, reduced the DADLE- induced ERK phosphorylation by 68%. ERK phosphorylation was inhibited by Bisindolyl-maleimide 1 (GF109203X), an inhibitor of PKC, and by pretreatment with PMA prior to DADLE. A GTP/GDP exchange assay was used to assess the potential role of Ras in the pathway leading to ERK phosphorylation; DADLE failed to stimulate GTP/GDP exchange in comparison to PMA. Additional studies showed that DADLE stimulated an increase in cfos mRNA; this was reduced by the inhibitor of MAPK/ERK kinase (MEK), PD98059. Therefore, in DOR-Ju.1 cells, DOR agonists stimulate ERK phosphorylation in a Ras independent and PKC-dependent manner; PTKs appear to be involved. MAPKs mediate the increase in cfos mRNA induced by DOR agonists.

Original languageEnglish (US)
Pages (from-to)48-57
Number of pages10
JournalJournal of Neuroimmunology
Volume94
Issue number1-2
DOIs
StatePublished - Feb 1 1999

Fingerprint

delta Opioid Receptor
Jurkat Cells
Phosphotransferases
Mitogen-Activated Protein Kinases
Phosphorylation
naltrindole
Guanosine Triphosphate
GTP-Binding Proteins
Gi-Go GTP-Binding Protein alpha Subunits
T-Lymphocytes
Messenger RNA
Opioid Peptides
Narcotic Antagonists
Pertussis Toxin
Protein Kinase Inhibitors
Concanavalin A
Protein-Tyrosine Kinases
Interleukin-2
Ala(2)-enkephalinamide-met
Peptides

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Neurology
  • Clinical Neurology

Cite this

Delta opioid receptors expressed by stably transfected jurkat cells signal through the map kinase pathway in a ras-independent manner. / Shahabi, Nahid A.; Daaka, Yehia; McAllen, Kathy; Sharp, Burt.

In: Journal of Neuroimmunology, Vol. 94, No. 1-2, 01.02.1999, p. 48-57.

Research output: Contribution to journalArticle

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abstract = "Delta opioid receptors (DOR) are G-protein coupled 7-transmembrane receptors (GPCR), expressed by thymic and splenic T cells, that modulate interleukin (IL)-2 production and proliferation in response to concanavalin A or crosslinking the TCR. Mitogen-activated protein kinases (MAPKs) are involved in mediating intracellular responses to TCR crosslinking. In addition, MAPKs can be activated by signaling cascades that are initiated by the release of G-proteins from GPCRs. To determine whether DORs expressed by T cells signal through the MAPKs, extracellular-regulated kinases (ERKs) 1 and 2, two delta opioid peptides, deltorphin and [D-Ala2, D-Leu5]- enkephalin (DADLE), were studied in Jurkat cells that had been stably transfected with DOR (DOR-Ju.1). These peptides rapidly and dose-dependently induced ERK phosphorylation; pretreatment with naltrindole (NTI), a selective DOR antagonist: abolished this. Pertussis toxin (PTX) also inhibited phosphorylation, indicating the involvement of the G(i/o) proteins. Herbimycin A, a protein tyrosine kinase (PTK) inhibitor, reduced the DADLE- induced ERK phosphorylation by 68{\%}. ERK phosphorylation was inhibited by Bisindolyl-maleimide 1 (GF109203X), an inhibitor of PKC, and by pretreatment with PMA prior to DADLE. A GTP/GDP exchange assay was used to assess the potential role of Ras in the pathway leading to ERK phosphorylation; DADLE failed to stimulate GTP/GDP exchange in comparison to PMA. Additional studies showed that DADLE stimulated an increase in cfos mRNA; this was reduced by the inhibitor of MAPK/ERK kinase (MEK), PD98059. Therefore, in DOR-Ju.1 cells, DOR agonists stimulate ERK phosphorylation in a Ras independent and PKC-dependent manner; PTKs appear to be involved. MAPKs mediate the increase in cfos mRNA induced by DOR agonists.",
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