title: Time course for early adaptive responses to ultraviolet B light in the epidermis of SKH-1 mice.
abstract: Hairless SKH-1 mice were exposed once to UVB light (180 mJ/cm2), and mechanistically important early adaptive responses in the epidermis were evaluated by immunohistochemical and morphological methods. Interrelationships in the time course for these UVB-induced responses were examined. The number of epidermal cells with DNA strand breaks (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells) or with thymine dimers increased to maximal levels within 30 min after UVB. The number of cells with DNA strand breaks located specifically in the basal layer of the epidermis was increased substantially by 3-30 min after UVB and gradually increased further over the next 5.5 hours. DNA strand breaks specifically in the basal layer of the epidermis were increased maximally at 6 h after UVB. The number of epidermal cells with DNA strand breaks or thymine dimers decreased markedly between 12 and 36 h. Pyrimidine (6-4) pyrimidone photodimers (6-4 photoproducts) in isolated epidermal DNA were increased immediately after irradiation of the mice with UVB and decreased markedly during the next 6 h. Exposure to UVB caused a rapid 8-fold increase in the number of epidermal cells with the DNA mismatch repair protein, MSH2 (within 30-60 min), and the level of MSH2-positive cells remained elevated for at least 48 h. These observations suggest a possible role of MSH2 in the repair of UVB-induced DNA damage. The number of epidermal cells with wild-type p53 protein started to increase at 1 h after UVB exposure and reached maximal levels by 8-12 h. The number of p53-positive cells fell markedly between 24 and 48 h. The time course for UVB-induced increases in the number of p53-positive cells was paralleled very closely by the time course for UVB-induced increases in the number of cells with p21(WAF1/CIP1), increases in morphologically distinct apoptotic sunburn cells, and decreases in the number of epidermal cells with bromodeoxyuridine (BrdUrd) incorporation into DNA. Although the start of UVB-induced increases in the number of p21(WAF1/CIP1)-positive cells was similar to that for the increase in p53-positive cells and very high levels of p21(WAF1/CIP1)-positive cells were observed at 8-12 h, maximal increases in p21(WAF1/CIP1)-positive cells were not achieved until 24 h after UVB irradiation (approximately 12 h after the peak value for p53). Myeloperoxidase-positive epidermal cells started to increase by 30 min after UVB exposure, and maximal numbers of myeloperoxidase-positive epidermal cells were observed at 2 h after UVB (18-fold higher than in nonirradiated control mice). An increased level of epidermal peroxidase enzyme activity in the epidermis was also observed from 1 to 24 h after exposure of the mice to UVB. Although neutrophil infiltration into the epidermis was not seen after exposure to UVB, neutrophil infiltration into the dermis (inflammatory response) was observed from 4 to 144 h after UVB exposure. In contrast to the marked inhibitory effect of UVB on BrdUrd incorporation into the DNA of epidermal cells observed at 8-12 h after UVB irradiation (>90% inhibition), BrdUrd incorporation into the DNA of epidermal cells was markedly increased (approximately 30-fold increase in the number of BrdUrd-positive cells) at 48 h after UVB exposure, and increases in epidermal cell layers and epidermal thickness (hyperplasia) were also observed. These later effects were associated with regeneration of the damaged epidermis.
authors.affiliation: Department of Chemical Biology, College of Pharmacy, Rutgers, The State University of New Jersey, Piscataway 08854-8020, USA.
authors.forename: Y P
authors.initials: YP
authors.lastname: Lu
authors.forename: Y R
authors.initials: YR
authors.lastname: Lou
authors.forename: P
authors.initials: P
authors.lastname: Yen
authors.forename: D
authors.initials: D
authors.lastname: Mitchell
authors.forename: M T
authors.initials: MT
authors.lastname: Huang
authors.forename: A H
authors.initials: AH
authors.lastname: Conney
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000528:radiation effects; D001205:Ascorbic Acid / Q000032:analysis; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000032:analysis; D004247:DNA / Q000737:chemistry / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects; D004268:DNA-Binding Proteins; D004817:Epidermis / Q000473:pathology / Q000528:radiation effects*; D005260:Female; D005978:Glutathione / Q000032:analysis; D020287:In Situ Nick-End Labeling; D051379:Mice; D008812:Mice, Hairless; D051718:MutS Homolog 2 Protein; D011518:Proto-Oncogene Proteins / Q000032:analysis; D011740:Pyrimidine Dimers / Q000032:analysis; D012867:Skin / Q000473:pathology / Q000528:radiation effects*; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000032:analysis; D014466:Ultraviolet Rays
chemical_list: C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D011740:Pyrimidine Dimers; D016159:Tumor Suppressor Protein p53; D004247:DNA; C497173:Msh2 protein, mouse; D051718:MutS Homolog 2 Protein; D005978:Glutathione; D001205:Ascorbic Acid