Immunohistochemical detection of induced expression of wild-type p53 tumor suppressor protein in the livers of rats treated with diethylnitrosamine.
Paraffin sections of formalin-perfused rat livers were stained immunohistochemically for p53. In livers from untreated rats, no p53 expression was observed. p53 expression was induced in a response to treatment with diethylnitrosamine 24 h prior to sacrifice. Staining for p53 was localized in the nucleus of perivenous hepatocytes. In serial sections p53-immunopositive areas were found to co-localize with increased expression of TUNEL-positive cells. Without formalin perfusion, the staining for p53 was uneven and often barely detectable. Perfusion with saline prior to formalin resulted in a rapid decrease in the detectability of p53, indicating rapid degradation of this protein under these conditions. We conclude that rapid fixation by formalin perfusion increases the detectability of p53 by immunohistochemical staining. This provides a convenient procedure for studying the response of wild-type p53 in rodent liver. This procedure is also suitable for in situ investigations on the degradation of p53 protein stabilized by DNA damage.
Martens
U
U
Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Stenius
U
U
D000818:Animals; D002273:Carcinogens / Q000494:pharmacology*; D002467:Cell Nucleus / Q000737:chemistry / Q000187:drug effects; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004052:Diethylnitrosamine / Q000494:pharmacology*; D005260:Female; D005557:Formaldehyde / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D016147:Genes, Tumor Suppressor / Q000235:genetics; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008099:Liver / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000096:biosynthesis / Q000187:drug effects*
D002273:Carcinogens; D016159:Tumor Suppressor Protein p53; D005557:Formaldehyde; D004052:Diethylnitrosamine; D004247:DNA