Research paper| Volume 14, ISSUE 1, P54-62, January 1997

Novel method to investigate kinetics of rat skin cells by means of an occlusive dressing method using bromodeoxyuridine

      This paper is only available as a PDF. To read, Please Download here.


      We developed a novel technique to detect S-phase skin cells by applying bromodeoxyuridine (BrdU) epicutaneously using an occlusive dressing (OD) method. BrdU was scarcely absorbed from the skin with a simple epicutaneous application, whereas the incorporation of BrdU was very well promoted with the use of our OD method. We applied BrdU on the backs of rats using this method and investigated the conditions required for an optimal response, with a special focus on the period of application, the concentration of BrdU used and vehicles suitable for the immunocytochemical staining of this agent. From these experiments, we were able to detemine that an application time of at least 60 min was necessary to label S-phase cells, a 2% concentration of BrdU was needed to obtain consistent labeling and aqueous vehicles are satisfactory solvents for BrdU preparations. Epidermal keratinocytes and S-phase cells in the upper portion of dermis were clearly labeled after either intraperitoneal injection of BrdU or after administration by means of the OD method. To ascertain whether this latter method could provide an effective alternative to intraperitoneal injection, we compared the labeling patterns of both methods with respect to the speed of migration of BrdU-labeled basal cells from the basal layer to the horny layer of epidermis. Using either of these two methods, basal keratinocytes were labeled immediately after administration. Three days after the first administration, BrdU-labeled cells were detected in the middle layer of the epidermis, but after 8 days, they were no longer evident in epidermal tissue. As another means of comparing both methods, we used antibody to proliferating cell nuclear antigen (PCNA) and compared the ratio of PCNA-positive basal cells to BrdU-labeled basal cells. The number of PCNA-positive cells was about 4.6 times greater than the number of BrdU-labeled basal cells by both methods. We concluded that the OD method could be used as a substitute for intraperitoneal injection in order to observe cell kinetics using bromodeoxyuridine.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Dermatological Science
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Bacchi CE
        • Gown AM
        Detection of cell proliferation in tissue sections.
        Brazilian J Med Biol Res. 1993; 26: 677-687
        • Bidmon H-J
        • Pitts JD
        • Bondi JV
        • Stumpf WE
        Estradiol distribution and penetration in rat skin after topical application, studied by high resolution autoradiography.
        His-tochemistry. 1990; 95: 43-54
        • Bravo R
        • Frank R
        • Blundell PA
        • Macdonald-Bravo H
        Cyclin/PCNA is the auxiliary protein of DNA poly-merase-δ.
        Nature. 1987; 326: 515-517
        • Dolbeare F
        • Gratzner H
        • Pallavicine MG
        • Gray JW
        Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine.
        in: Proc Natl Acad Sci USA. 69. 1983: 5573-5577
        • Furukawa F
        • Imamura S
        • Fujita M
        • Kinoshita K
        • Yoshitake K
        • Brown WR
        • Norris DA
        Immunohistochemical localization of proliferating cell nuclear antigen/cyclin in human skin.
        Arch Dermatol Res. 1992; 284: 86-91
        • Geary WA
        • Cooper PH
        Proliferating cell nuclear antigen (PCNA) in common epidermal lesions. An immunohisto-chemical study of proliferating cell populations.
        J Cutan Pathol. 1992; 19: 458-468
        • Goz B
        The effects of incorporation of 5-halogenated deoxyuridines into the DNA of eukaryotic cells.
        Pharmacol Rev. 1978; 29: 249-272
        • Gratzner HG
        Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: a new reagent for detection of DNA replication.
        Science. 1982; 218: 474-475
        • Hamada S
        A double labeling technique combining 3H- thymidine autoradiography with BrdUrd immunocyto-chemistry.
        Acta Histochem Cytochem. 1985; 18: 267-270
        • Heartlein MW
        • O'Neill JP
        • Pal BC
        • Preston RJ
        The induction of specific locus mutations and sister-chromatid exchanges by 5-bromo- and 5-chlorodeoxyuridine.
        Mutat Res. 1982; 92: 441-446
        • Hegazy MAH
        • Fowler JF
        Cell population kinetics of plucked and unplucked mouse skin.
        Cell Tissue Kinet. 1973; 6: 17-33
        • Hoshino T
        • Nagashima T
        • Murovic J
        • Levin EM
        • Levin VA
        • Rupp SM
        Cell kinetic studies of in situ human brain tumors with bromodeoxyuridine.
        Cytometry. 1985; 6: 627-632
        • Isik FF
        • Ferguson M
        • Yamanaka E
        • Gordon D
        Proliferating cell nuclear antigen. A marker for cell proliferation in autopsy tissues.
        Arch Pathol Lab Med. 1992; 116: 1142-1146
        • Lacy ER
        • Kuwayama H
        • Cowart KS
        • King JS
        • Deutz AH
        • Sistrunk S
        A rapid, accurate, immunohistochemical method to label proliferating cells in the digestive tract.
        Gastroenterology. 1991; 100: 259-262
        • Mathews MB
        • Bernstein RM
        • Garrels JI
        Identity of the proliferating cell nuclear antigen and cyclin.
        Nature. 1984; 23: 505-522
        • Morimoto Y
        • Saga K
        • Bando M
        • Takahashi M
        In-vitro DNA synthesis of keratinocytes in normal human skin, psoriasis, seborrhoeic keratosis, Bowen's disease and basal cell carcinoma.
        Br J Dermatol. 1991; 125: 9-13
        • Morstyn G
        • Hsu SM
        • Gratzner HG
        • Russo A
        • Mitchell JB
        Bromodeoxyuridine in tumors and chromosomes detected with a monoclonal antibody.
        J Clin Invest. 1983; 72: 1844-1850
        • Raza A
        • Spiridonidis C
        • Ucar K
        • Mayers G
        • Bankert R
        • Prisler HD
        Double labeling of S-phase murine cells with bromodeoxyuridine and a second DNA-specific probe.
        Cancer Res. 1985; 45: 2283-2287
        • San Sebastian JR
        • O'Neill JP
        • Pal BC
        • Preston RJ
        The induction of chromosome aberrations, sister chromatid exchanges, and specific locus mutation in Chinese hamster ovary cells by 5-bromodeoxyuridine.
        Cytogenet Cell Genet. 1980; 28: 47-54
        • Thiry M
        • Dombrowicz D
        Anti-bromodeoxyuridine monoclonal antibody: an alternative tool for the identification of replicated DNA at the electron microscopic level.
        Biol. Cell. 1988; 62: 99-102
        • Tsuyama S
        • Yang DH
        • Kanae T
        • Kasamo H
        • Murata F
        Immunoelectron microscopic labeling of digestive tract stem cells by means of bromodeoxyuridine antibody.
        Acta Histochem. Cytochem. 1994; 27: 339-345
        • Yu CC-W
        • Woods AL
        • Levison DA
        The assessment of cellular proliferation by immunohistochemistry: a review of currently available methods and their applications.
        Histochem J. 1992; 24: 121-131