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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1989 Sep;80(9):826–832. doi: 10.1111/j.1349-7006.1989.tb01722.x

In situ Freezing of the Rat Urinary Bladder: DNA Adduct Formation in the Bladder Epithelium Demonstrated by 32P‐Postlabeling Assay

Ryohei Hasegawa 1,✉, Katsumi Yamashita 3,†, Kazushige Morimoto 2, Fumio Furukawa 1, Kazuhiro Toyoda 1, Naoshi Shimoji 1, Michihito Takahashi 1, Yuzo Hayashi 1
PMCID: PMC5917847  PMID: 2513299

Abstract

In situ freezing of the urinary bladder has been demonstrated to exert tumor‐initiating potential in two‐stage urinary bladder carcinogenesis in the rat. In the present experiment, DNA modification was examined after in situ freezing of the whole urinary bladder performed by pinching with frozen forceps at —15°C or —30°C for 2 s. The 32P‐postlabeling analysis revealed at least 2 DNA adducts in the epithelial cells of the urinary bladder collected 3 days after freezing. Single‐strand breaks of DNA were also found by means of the alkaline elution assay in the bladder epithelium collected 10 min after freezing. Thus, the previously demonstrated tumor‐initiating activity of in situ freezing in urinary bladder carcinogenesis was revealed to be associated with substantial DNA damage and adduct formation.

Keywords: In situ freezing, Urinary bladder carcinogenesis, 32P‐Postlabeling, DNA adducts, Rat

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