Quantitative detection of ultraviolet light-induced photoproducts in mouse skin by immunohistochemistry

Jpn J Cancer Res. 1995 Nov;86(11):1041-8. doi: 10.1111/j.1349-7006.1995.tb03018.x.

Abstract

UVB-induced cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4)photoproducts [(6-4)photoproducts] in mouse skin DNA were quantitatively measured using an immunohistochemical approach with a computer-aided color image analyzer. The skins of the C3H/HeN mice were irradiated with ultraviolet B (UV-B, 280-320 nm), and processed to give conventional formalin-fixed, paraffin-embedded histologic sections. Routine immunohistochemistry clearly demonstrated a dose-dependent induction of both photoproducts. CPDs were detectable at doses > or = 125 J/m2, while for (6-4)photoproducts, the minimal dose at which they were detectable was 250 J/m2 in the present study. A time course study showed that the repair of (6-4)photoproducts was more rapid than that of CPDs, and that epidermal cells had a higher capacity for their removal than dermal cells. About half of the (6-4)photoproducts were excised within the first 24 h after the irradiation, and the process was essentially complete by 72 h. In contrast, there was no apparent removal (less than 10%) of CPDs in the first 24 h and they only completely disappeared from the epidermal cells at 120 h after irradiation. The effect of DNA dilution due to increased turnover of epidermal cells after UV-B irradiation was evaluated by quantitative immunohistochemical measurement of the time course of bromodeoxy-uridine (BrdUrd) incorporated into nuclei at 2 days post irradiation when the proliferation reaches a peak. The removal of photoproducts was more marked than the decrease in BrdUrd staining. Our results suggest that mouse skin cells can repair both (6-4)photoproducts and CPDs, but with considerably lower efficiency, especially in the latter case, then human or monkey skin cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Division
  • Color
  • DNA / radiation effects*
  • DNA Damage*
  • DNA Repair
  • Epidermis / chemistry
  • Epidermis / pathology
  • Epidermis / radiation effects
  • Female
  • Fibroblasts / chemistry
  • Fibroblasts / pathology
  • Fibroblasts / radiation effects
  • Image Processing, Computer-Assisted
  • Immunoenzyme Techniques
  • Mice
  • Mice, Inbred C3H
  • Photochemistry
  • Pyrimidine Dimers / analysis*
  • Skin / chemistry
  • Skin / radiation effects*
  • Ultraviolet Rays*

Substances

  • Pyrimidine Dimers
  • pyrimidine-pyrimidone dimer
  • DNA