Using Laser Scanning Cytometry to Measure PPAR-Mediated Peroxisome Proliferation and β Oxidation

Laser scanning cytometry (LSC) is a new technology that combines the properties and advantages of flow cytometry (FC) and immunohistochemistry (IHC), thus providing qualitative and quantitative information on protein expression with the additional perspective provided by cell and tissue localization...

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Bibliographic Details
Published in:	Toxicologic pathology Vol. 33; no. 1; pp. 86 - 91
Main Authors:	Pruimboom-Brees, Ingrid M., Brees, Dominique J. J. E., Shen, Amy C., Keener, Mary, Francone, Omar, Amacher, David E., Loy, James K., Kerlin, Roy L.
Format:	Journal Article
Language:	English
Published:	Thousand Oaks, CA SAGE Publications 01-01-2005 Sage
Subjects:	Acyl-CoA Oxidase - metabolism Animals Biological and medical sciences Catalase - metabolism Dose-Response Relationship, Drug Female Laser Scanning Cytometry Liver - metabolism Male Medical sciences Oxidation-Reduction Palmitoyl Coenzyme A - metabolism Peroxisome Proliferator-Activated Receptors - metabolism Peroxisome Proliferators - toxicity Peroxisomes - drug effects Peroxisomes - metabolism Rats Rats, Sprague-Dawley Reproducibility of Results Toxicology Peroxisome β-oxidation Acyl CoA oxidase proliferation PTS1 Cell proliferation Enzyme Acyl-CoA oxidase Peroxisome proliferator Cytometry Laser Oxidation Oxidoreductases Peroxisome proliferator activated receptor
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Summary:	Laser scanning cytometry (LSC) is a new technology that combines the properties and advantages of flow cytometry (FC) and immunohistochemistry (IHC), thus providing qualitative and quantitative information on protein expression with the additional perspective provided by cell and tissue localization. Formalin-fixed, paraffin embedded liver sections from rats exposed to a Peroxisome Proliferator Activated Receptor (PPAR) agonist were stained with antibodies against peroxisomal targeting signal-1 (PTS-1) (a highly conserved tripeptide contained within all peroxisomal enzymes), Acyl CoA oxidase (AOX) (the rate limiting enzyme of peroxisomal β oxidation), and catalase (an inducible peroxisomal antioxidant enzyme) to evaluate peroxisomal β oxidation, oxidative stress, and peroxisome proliferation. The LSC showed increased AOX, catalase, and PTS-1 expression in centrilobular hepatocytes that correlated favorably with the microscopic observation of centrilobular hepatocellular hypertrophy and with the palmitoyl CoA biochemical assay for peroxisomal β oxidation, and provided additional morphologic information about peroxisome proliferation and tissue patterns of activation. Therefore, the LSC provides qualitative and quantitative evaluation of peroxisome activity with similar sensitivity but higher throughput than the traditional biochemical methods. The additional benefits of the LSC include the direct correlation between histopathologic observations and peroxisomal alterations and the potential utilization of archived formalin-fixed tissues from a variety of organs and species.
ISSN:	0192-6233 1533-1601
DOI:	10.1080/01926230590881817