Quantitative detection of ultraviolet light-induced photoproducts in mouse skin by immunohistochemistry.
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.
Qin
X
X
Department of Pathology, Faculty of Medicine, University of Tokyo.
Zhang
S
S
Oda
H
H
Nakatsuru
Y
Y
Shimizu
S
S
Yamazaki
Y
Y
Nikaido
O
O
Ishikawa
T
T
D000818:Animals; D002455:Cell Division; D003116:Color; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair; D004817:Epidermis / Q000737:chemistry / Q000473:pathology / Q000528:radiation effects; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000473:pathology / Q000528:radiation effects; D007091:Image Processing, Computer-Assisted; D007124:Immunoenzyme Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D010777:Photochemistry; D011740:Pyrimidine Dimers / Q000032:analysis*; D012867:Skin / Q000737:chemistry / Q000528:radiation effects*; D014466:Ultraviolet Rays
D011740:Pyrimidine Dimers; C058406:pyrimidine-pyrimidone dimer; D004247:DNA