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Role of Protein Kinase C α in Nerve Growth Factor-Induced Arachidonic Acid Release from PC12 Cells

Authors

  • Wen-Hua Zheng,

    1. Section on Growth Factors, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, U.S.A.
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  • Donald W. Fink Jr.,

    1. Section on Growth Factors, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, U.S.A.
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    • The present address of Dr. D. W. Fink, Jr., is Division of Cytokine Biology, Laboratory of Cell Biology/Neurotrophic Factors, Food and Drug Administration, Bethesda, MD 20892, U.S.A.

  • Gordon Guroff

    Corresponding author
    1. Section on Growth Factors, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, U.S.A.
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Address correspondence and reprint requests to Dr. G. Guroff at Section on Growth Factors, National Institute of Child Health and Human Development, Building 49, Room 5A64, Bethesda, MD 20892, U.S.A.

Abstract

Abstract: Nerve growth factor (NGF) increases arachidonic acid (AA) release by PC12 pheochromocytoma cells. To explore the role of protein kinase C (PKC) in this action of NGF, PKC was down-regulated by long-term treatment of the cells with phorbol 12-myristate 13-acetate (PMA). Such prolonged exposure to PMA (1 µM) resulted in the inhibition of NGF-induced AA release. Moreover, pretreatment of PC12 cells with the protein kinase inhibitor staurosporine or with calphostin C, a specific inhibitor of PKC, also blocks the increase of AA release induced by NGF. These data, as well as that PMA alone can induce AA release in PC12 cells, suggest that PKC is necessary for NGF-induced AA release. Immunoblot analysis of whole cell lysates by using antibodies against various PKC isoforms revealed that our PC12 cells contained PKCs α, δ, ε, and ζ. PMA down-regulation depleted PKCs α, δ, and ε, and partially depleted ζ. To see which isoform was involved in NGF-induced AA release, an isoform-specific PKC inhibitor was used. GO 6976, a compound that inhibits PKCs α and β specifically, blocked NGF-induced AA release. In addition, thymeleatoxin, a specific activator of PKCs α, β, and γ, induced AA release from PC12 cells in amounts comparable with those seen with NGF. Taken together, these data suggest that PKC α plays a role in NGF-induced AA release.

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