Flow cytometric assessment of peroxisome proliferation from frozen liver of fibrate-treated monkeys

International Journal of Toxicology
Puntipa KwanyuenNeal F Cariello

Abstract

Multiple methods currently exist for the assessment of peroxisome proliferation, including gene expression, enzyme activity, immunolabeling coupled with image analysis, and electron microscopy. This study describes a novel flow cytometric method to efficiently quantify peroxisome proliferation in cells from frozen livers. Frozen livers from cynomolgus monkeys treated with ciprofibrate at doses of 0, 3, 30, 150, and 400 mg/kg/day for 15 days were mechanically disaggregated using an automated dispersion method. The resulting cell suspensions were labeled using an allophycocyanin (APC)-conjugated antibody directed against peroxisomal membrane protein 70 (PMP70). Statistically significant increases in mean fluorescence intensity were observed from animals dosed at 30, 150, and 400 mg/kg/day compared to control. Parallel comparisons using electron microscopy and immunofluorescence microscopy suggest that flow cytometry may be an alternative to electron microscopy in determinations of peroxisome proliferation. Flow cytometric analysis of freshly isolated hepatocytes and frozen liver from rats treated with fenofibrate at 200 mg/kg/day for 10 days showed the flow cytometric method could detect peroxisome proliferation in both species. ...Continue Reading

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Citations

Jul 27, 2007·Cell Biochemistry and Biophysics·Jay E Brenman, Brenda R S Temple
Oct 23, 2010·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Kuok Teong OngDouglas G Mashek
Sep 17, 2009·Journal of Applied Toxicology : JAT·Frederikke Lihme EgerodMartin B Oleksiewicz

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Methods Mentioned

BETA
electron microscopy
flow cytometry
Fluorescence
electrophoresis
Infrared Imaging
electron

Software Mentioned

Image
CellQuest
Medimachine
WinList
Quantum Dots
JMP
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