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Research ArticleArticle

Neuropeptide-Y Stimulation of Extracellular Signal-Regulated Kinases in Human Erythroleukemia Cells

Susanne Keffel, Martina Schmidt, Angela Bischoff and Martin C. Michel
Journal of Pharmacology and Experimental Therapeutics December 1999, 291 (3) 1172-1178;
Susanne Keffel
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Martina Schmidt
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Angela Bischoff
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Martin C. Michel
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Abstract

We have used human erythroleukemia (HEL) cells to investigate distal signaling mechanisms of neuropeptide-Y (NPY) receptors. NPY did not activate phospholipase D, determined as a phosphatidylethanol formation, or protein kinase C (PKC) determined enzymatically as a translocation to the plasma membrane. However, NPY caused a rapid (already maximal after 30 s) and concentration-dependent (maximum at 10–100 nM) activation of extracellular signal-regulated kinase (ERK) as assessed by immunoblotting with epitope-specific, antiphosphotyrosine antibodies and in some cases enzymatically. ERK activation by 100 nM NPY was abolished by the Y1 NPY receptor antagonist BIBP 3226 (1 μM), pertussis toxin treatment (100 ng ml−1 overnight), the mitogen-activated protein kinase (MAPK) kinase inhibitor PD 98059 (100 μM), and the phosphatidylinositol-3-kinase inhibitor wortmannin (100 nM). Whereas the PKC inhibitor staurosporine (3 μM) inhibited ERK activation by NPY, the chemically distinct PKC inhibitors calphostin C (3 μM), Gö 6976 (3 μM), and bisindolylmaleimide I (3 μM) did not. NPY did not activate other MAPK such as jun N-terminal kinase or p38 MAPK. We conclude that NPY does not activate phospholipase D, PKC, jun N-terminal kinase, or p38 MAPK in HEL cells. However, NPY activates ERK by a pathway involving Y1 receptors, pertussis toxin-sensitive G proteins, and phosphatidylinositol-3-kinase, whereas PKC may not be involved. Staurosporine may have PKC-independent effects on ERK activation.

Footnotes

  • Send reprint requests to: Dr. Martin C. Michel, Nephrologisches Labor IG 1, Klinikum, 45122 Essen, Germany. E-mail: martin.michel{at}uni-essen.de

  • ↵1 This work was supported in part by a grant from the Deutsche Forschungsgemeinschaft (Mi 294/5-2).

  • Abbreviations:
    NPY
    neuropeptide-Y
    CHO
    Chinese hamster ovary
    ERK
    extracellular signal-regulated kinase
    HEL
    human erythroleukemia
    JNK
    Jun N-terminal kinase
    MAPK
    mitogen-activated protein kinase
    PKC
    protein kinase C
    PMA
    phorbol-12-myristate-13-acetate
    MEK
    mitogen-activated protein/extracellular signal-related kinase kinase
    • Received March 3, 1999.
    • Accepted August 18, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 291 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 291, Issue 3
1 Dec 1999
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Research ArticleArticle

Neuropeptide-Y Stimulation of Extracellular Signal-Regulated Kinases in Human Erythroleukemia Cells

Susanne Keffel, Martina Schmidt, Angela Bischoff and Martin C. Michel
Journal of Pharmacology and Experimental Therapeutics December 1, 1999, 291 (3) 1172-1178;

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Research ArticleArticle

Neuropeptide-Y Stimulation of Extracellular Signal-Regulated Kinases in Human Erythroleukemia Cells

Susanne Keffel, Martina Schmidt, Angela Bischoff and Martin C. Michel
Journal of Pharmacology and Experimental Therapeutics December 1, 1999, 291 (3) 1172-1178;
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