Flow Cytometric Assessment of Peroxisome Proliferation from Frozen Liver of Fibrate-Treated Monkeys
| Autor: | Don Creech, Puntipa Kwanyuen, Neal F. Cariello, J. Greg Falls, Sam M. Witherspoon, Heidi M. Colton |
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| Rok vydání: | 2006 |
| Předmět: |
Male
0301 basic medicine Peroxisome Proliferation Cell Separation 010501 environmental sciences Biology Toxicology 01 natural sciences Flow cytometry Rats Sprague-Dawley Clofibric Acid 03 medical and health sciences Immunolabeling Fenofibrate Microscopy Electron Transmission Peroxisomes medicine Animals 0105 earth and related environmental sciences Cryopreservation Allophycocyanin Dose-Response Relationship Drug 030102 biochemistry & molecular biology medicine.diagnostic_test Fibric Acids Peroxisome Flow Cytometry Molecular biology Enzyme assay Rats Macaca fascicularis Liver Hepatocytes biology.protein Feasibility Studies Peroxisome Proliferators Ciprofibrate medicine.drug |
| Zdroj: | International Journal of Toxicology. 25:41-47 |
| ISSN: | 1092-874X 1091-5818 |
| DOI: | 10.1080/10915810500488395 |
| Popis: | 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. The research described here demonstrates the feasibility of applying flow cytometry for the detection of peroxisome proliferation. |
| Databáze: | OpenAIRE |
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