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Research ArticleArticle

Characterization of Pardaxin-Induced Dopamine Release from Pheochromocytoma Cells: Role of Calcium and Eicosanoids

Saleh Abu-Raya, Eugenia Bloch-Shilderman, Peter I. Lelkes, Victoria Trembovler, Esther Shohami, Yehuda Gutman and Philip Lazarovici
Journal of Pharmacology and Experimental Therapeutics February 1999, 288 (2) 399-406;
Saleh Abu-Raya
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Eugenia Bloch-Shilderman
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Peter I. Lelkes
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Victoria Trembovler
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Esther Shohami
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Yehuda Gutman
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Philip Lazarovici
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Abstract

Pardaxin, an excitatory neurotoxin, induced dopamine release from pheochromocytoma (PC12) cells both in the presence and absence of extracellular calcium ([Ca]o). In the presence of extracellular calcium, nifedipine, an l-type calcium channel blocker, did not affect dopamine release, whereas 1,2-bis (2-aminophenoxy) ethane N,N,N′N′-tetra-acetic acid (BAPTA), a chelator of cytosolic calcium, and dantrolene, a blocker of calcium release from intracellular stores, inhibited only partially (30–40%) pardaxin-induced dopamine release. In the absence of [Ca]o, BAPTA and dantrolene were ineffective. Pardaxin stimulated the arachidonic acid (AA) cascade in PC12 cells independently of [Ca]o. The phospholipase inhibitors mepacrine and bromophenacyl bromide inhibited both pardaxin-induced AA release and pardaxin-induced dopamine release. Dopamine release induced by pardaxin also was blocked by the lipoxygenase inhibitors nordihydroguaiaretic acid, esculetin, and 2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-1,4-benzoquinone. Under these conditions, a parallel reduction in 5-hydroxyeicosatetranoic acid release also was observed. Suppression of pardaxin-induced dopamine release by inhibitors of phospholipase A2 and lipoxygenase was more pronounced in calcium-free medium. These results indicate the involvement of the lipoxygenase pathway in pardaxin-induced dopamine release and suggest the use of this toxin as a novel pharmacological tool for investigating the mechanism of calcium-independent neurotransmitter release.

Footnotes

  • Send reprint requests to: Philip Lazarovici, Department of Pharmacology and Experimental Therapeutics, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, P.O. Box 12065, Jerusalem 91120, Israel.

  • ↵1 Affiliated with the David R. Bloom Center for Pharmacy at the Hebrew University.

  • Abbreviations:
    PX
    pardaxin
    BPB
    4-bromophenacyl bromide
    CCH
    carbachol
    [Ca]o
    extracellular calcium
    [Ca]i
    cytosolic-free calcium
    Dan
    dantrolene
    NDGA
    nordihydroguaiaretic acid
    AA861
    2-(12-hydroxydodeca-5,10-diynyl)-3,5,6-trimethyl-1,4-benzoquinone
    AA
    arachidonic acid
    5-HETE
    5-hydroxyeicosatetranoic acid
    Fura-2-AM
    acetoxymethyl ester of Fura-2
    DMEM
    Dulbecco’s modified Eagle’s medium
    PC12
    pheochromocytoma cells
    BAPTA
    1,2-bis(2-aminophenoxy)ethane-N,N,N′N′-tetra-acetic acid
    PLA2
    phospholipase A2
    IP3
    inositol 1,4,5-trisphosphate
    • Received October 1, 1997.
    • Accepted August 5, 1998.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 288 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 288, Issue 2
1 Feb 1999
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Research ArticleArticle

Characterization of Pardaxin-Induced Dopamine Release from Pheochromocytoma Cells: Role of Calcium and Eicosanoids

Saleh Abu-Raya, Eugenia Bloch-Shilderman, Peter I. Lelkes, Victoria Trembovler, Esther Shohami, Yehuda Gutman and Philip Lazarovici
Journal of Pharmacology and Experimental Therapeutics February 1, 1999, 288 (2) 399-406;

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Research ArticleArticle

Characterization of Pardaxin-Induced Dopamine Release from Pheochromocytoma Cells: Role of Calcium and Eicosanoids

Saleh Abu-Raya, Eugenia Bloch-Shilderman, Peter I. Lelkes, Victoria Trembovler, Esther Shohami, Yehuda Gutman and Philip Lazarovici
Journal of Pharmacology and Experimental Therapeutics February 1, 1999, 288 (2) 399-406;
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