Peroxisomes - or microbodies - are organelles present in nearly all cells. Interest stems not only from their various biological and cell physiological roles but also because several genetic diseases are known with deficient peroxisomal functions. In response to several drugs or pesticides, peroxisomes start to proliferate. Due to this property they may also be used as toxicological indicators. Designed for use at the lab bench, this manual is ideal for everyone involved in peroxisome research or concerned with hepatotoxicity testing of new compounds.

I: Isolation and Characterization of Peroxisomes

General Indroduction to Isolation and Characterization of Peroxisomes
Experiment 1. Preparation and Purification of Peroxisomes; Subfractionation of Purified Peroxisomes; Acyl-CoA Oxidase Activity Measurements
Experiment 2. Immunoblotting of Peroxisomal Proteins with Monospecific Antibodies
Experiment 3. Morphology of Peroxisomes in Light- and Electron Microscopy
II: Molecular Biology
Experiment 4. Northern Blotting Analysis of Rat Liver mRNA Encoding for Peroxisomal Acyl-CoA Oxidase
Experiment 5. Transient Co-transfection Assay: Measure of the Chloramphenicol Acetyl Transferase (CAT) Activity in Cytosol Extracts from Transfected Cells
Experiment 6. Synthesis of Oligonucleotides Used as Probes; Purification by HPLC
Experiment 7. Computer Analysis of DNA and Protein Sequences
III: Toxicology and Pharmacology
Experiment 8. Cytochrome P450 Isoenzyme Induction Pattern in Rats Treated with Peroxisome Proliferators and Classical Inducers
Experiment 9. Measurement of the in Vitro Glucuronidation of Peroxisome Proliferator Carboxylic Acids by Liver Microsomes and by Genetically Modified V 79 Cell Line
Experiment 10. Molecular Modeling and Drug Design: Application to Some Peroxisome Proliferator Agents
IV: Cell Culture and Genetic Diseases
Experiment 11. Fao Cell Line as a Model for the Study of the Effect of Peroxisome Proliferators on Cellular Functions
Experiment 12. Measurement of Dihydroxyacetone-Phosphate Acyl-Transferase (DHAP-AT) Activity in Liver Peroxisomes and Cell Lines
Experiment 13. In Vitro Transformation of Human Fibroblasts with SV40 T Antigen (Lipofection)
Experiment 14. Somatic Cell Hybridization as a Tool for Genetic Analysis of Peroxisomal Activities
Supplement.