Products of the phospholipase A2 pathway mediate the dihydrorhodamine fluorescence response evoked by endogenous and exogenous peroxynitrite in PC12 cells

Letizia Palomba, Piero Sestili, Andrea Guidarelli, Clara Sciorati, Emilio Clementi, Mara Fiorani, Orazio Cantoni

Research output: Contribution to journalArticle

Abstract

A short-term exposure of PC12 cells to tert-butylhydroperoxide promotes a rapid oxidation of dihydrorhodamine sensitive to nitric oxide synthase inhibitors and peroxynitrite scavengers. This response was not directly caused by peroxynitrite, but rather appeared to be mediated by peroxynitrite-dependent activation of phospholipase A2. The following lines of evidence support this inference: (i) the peroxynitrite-dependent dihydrorhodamine fluorescence response was blunted by low concentrations of two structurally unrelated phospholipase A2 inhibitors; (ii) under similar conditions, the phospholipase A2 inhibitors prevented release of arachidonic acid; (iii) low levels of arachidonic acid restored the dihydrorhodamine fluorescence response in nitric oxide synthase- as well as phospholipase A2-inhibited cells; (iv) the dihydrorhodamine fluorescence response induced by authentic peroxynitrite was also blunted by phospholipase A2 inhibitors and restored upon addition of reagent arachidonic acid. We conclude that endogenous, or exogenous, peroxynitrite does not directly oxidize dihydrorhodamine in intact cells. Rather, peroxynitrite appears to act as a signalling molecule promoting release of arachidonic acid, which in turn leads to formation of species causing the dihydrorhodamine fluorescence response. Copyright (C) 2000 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)783-789
Number of pages7
JournalFree Radical Biology and Medicine
Volume29
Issue number8
DOIs
Publication statusPublished - Oct 15 2000

Keywords

  • Arachidonic acid
  • Dihydrorhodamine fluorescence response
  • Free radicals
  • Nitric oxide
  • Peroxynitrite
  • Tert-butylhydroperoxide

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

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