| Ornithine decarboxylase: Expression and regulation in rat brain and in transgenic mice | ||
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Rats were anesthetized with pentobarbital (30 mg/kg, i.p.) and perfused transcardially first with 100 ml of saline followed by a mixture of 4 % paraformaldehyde and 0.1 % picric acid in 0.1 M phosphate buffered saline (PBS), pH 7.3, for 4 – 5 min. After perfusion the brains were excised and further fixed by immersion in the same solution for 60 min. The samples were cryoprotected with 15 % sucrose in PBS before sectioning in the cryostat. Immunocytochemistry was carried out using ABC-method. Primary antibodies were rabbit antiserum against mouse ODC (Laitinen et al. 1986) and rabbit antiserum against rat antizyme (Matsufuji et al. 1990). Diaminobenzidine was used as a chromogen. After staining sections were dehydrated, mounted and examined with Nikon Microphot-FXA microscope. Controls included omission of the primary or secondary antibodies and preabsorption of the primary antibodies with respective proteins.