Year: 2002
Type: article
Abstract: ReceivedJune12，2002 Abstmct：ThemajortypesofDNAdamageinducedbysunlightintheskinareDNA photoproducts，SuCh as cyclobutane pyrimidine dimers(CPDs)，(6-4)photoproducts(6-4PPs)andDewarisomersof6-4PPs．Asensitivemethodforquantitatingandvisualizing eachtypeofDNAphotoproductinducedbybiologica11yrelevantdosesofUVorsunlight is essentialto characterize DNA photoproducts and their biologlCal effects．We have established monoclonal antibodies specific for CPDs，6-4PPs or Dewarisomers．Those antibodiesallowonetoquantitatephotoproductsinDNApurifiedfromculturedcellsor fromtheskinepidermisuslnganenZyme-linkedimmunosorbentassay．Onecanalsouse those specific antibodies within situlaser cytometry to visualizeand measure DNA photoproductsin culturedcells orin theskin，uSlngindirectimmunofluorescenceanda laser-SCannlng COnfocal microscope．Thislatter method allows us to reconstruct threedimensionalimages of nuclei containlng DNA photoproducts，and to simultaneously examine DNA photoproducts and histologyin multilayered epidermis．Using those techniques，One Can determine theinductionand repairofthese three distinct types of DNA photoproductsin cultured cells andin the skin exposed to sublethal or SuberythematousdosesofUVorsolarsimulatedradiation・Asexamplesoftheutilityof these techniques and antibodies，We describe the DNA repalr kinetics followlng irradiationofhumancellnucleiand thephotoprotectiveeffectofmelaninagainstDNA photoproductsinculturedpigrlentedcellsandinhumanepidermis． Key words‥Cyclobutane pyrimidine dimer，(6-4)photoproduct，Monoclonal antibody， ELISA，Immunofluorescence
Authors: Toshio Mori, Nobuhiko Kobayashi
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W362164140