Release of the lipid peroxidation marker 8-epi-prostaglandin F from isolated gill pavement cells

Eric G. Spokas, Scott Harshman, Glenn M. Cohen, Chen Jiang, Jaime M. Levine, Ana R. Rodriguez, Jon Foglein, Bernd W. Spur

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4 Scopus citations


The aim of the present study was to evaluate oxidative injury in gill pavement cells (GPCs) from fathead minnow (Pimephales promelas) using F 2-isoprostane (F2-iP) release as an index of lipid peroxidation. Cells were isolated from pooled gill tissue by collagenase treatment, mechanical sieving, and Percoll® density gradient centrifugation. Baseline levels of 8-epi-prosta-glandin F (8-epi-PGF ) were measured by incubating GPCs in physiological buffer (106 cells/ml) and enzyme immunoassay. After 60 min, the amount of immunoreactive 8-epi-PGF (ir8-epi-PGF ) in control medium ranged from 1,374 to 5,515 pg/ml. Lead nitrate, 0.6 to 120 μM, did not influence ir8-epi-PGF release, whereas FeCl3 stimulated release at 500 μM but not at 5 μM. Incubation medium was extracted for acidic lipids and analyzed by liquid chromatography/mass spectrometry/electrospray ionization. A compound in the medium exhibited a retention time on reverse-phase high-performance liquid chromatography nearly identical to that of synthetic 8-epi-PGF The mass spectrum taken from the total ion chromatogram from 14.8 to 15.1 min contained a prominent ion at mlz 353, as expected for the molecular ion of 8-epi-PGF. Similar results were obtained with tissue subjected to base hydrolysis. Mass spectra of extracted ion chromatograms obtained with gill extracts and authentic standard showed a close correspondence of fragment ions, providing definitive evidence for production and storage of F2-iPs by fish gills. In summary, F2-iP release occurs during lipid peroxidation injury to fish gill epithelium, and its measurement may facilitate aquatic toxicology studies of metallic and nonmetallic contaminants.

Original languageEnglish (US)
Pages (from-to)1569-1575
Number of pages7
JournalEnvironmental Toxicology and Chemistry
Issue number7
StatePublished - Jul 2008

All Science Journal Classification (ASJC) codes

  • Environmental Chemistry
  • Health, Toxicology and Mutagenesis


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