title: In vivo imaging demonstrates ATP release from murine keratinocytes and its involvement in cutaneous inflammation after tape stripping.
abstract: Adenosine 5'-triphosphate (ATP) release from keratinocytes has been observed in various stress models in vitro, but studies demonstrating epidermal ATP release in vivo are limited. To visualize extracellular ATP (eATP) in vivo, we developed enhanced green-emitting luciferase immobilized on agarose beads (Eluc-agarose). Subcutaneous injection of Eluc-agarose together with ATP into the dorsal skin of BALB/c mice following intraperitoneal luciferin injection produced detectable and measurable bioluminescence using an in vivo imaging system. Using Eluc-agarose, we demonstrated in vivo that bright bioluminescence was observed from 1 to 20 minutes after repeated tape stripping of murine skin. This bioluminescence was suppressed by the local administration of apyrase. Eluc-agarose bioluminescence was observed only in tape-stripped skin with transepidermal water loss (TEWL) between 100 and 140 g m(2) h(-1), indicating a loss of bioluminescence with excessive tape stripping (TEWL>140 g m(-2) h(-1)). Histologically, tape-stripped skin with detectable eATP had a viable epidermis and a subepidermal neutrophil infiltrate, and administration of apyrase reduced the inflammatory infiltrate. Neither a viable epidermis nor an upper dermal neutrophil infiltrate was observed after excessive tape stripping. These results suggest that tape stripping prompts ATP release from viable keratinocytes, which facilitates inflammatory cell migration. Eluc-agarose may be useful in the in vivo detection of eATP in murine models of skin diseases.
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Toshiya
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kimura
authors.affiliation: Photometry and Radiometry Division, National Metrology Institute of Japan, Tsukuba, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Niwa
authors.affiliation: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmiya
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Fujimura
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Kenshi
authors.initials: K
authors.lastname: Yamasaki
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan. Electronic address: saiba@med.tohoku.ac.jp.
authors.forename: Setsuya
authors.initials: S
authors.lastname: Aiba
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001081:Apyrase / Q000494:pharmacology; D018925:Chemokines / Q000235:genetics / Q000276:immunology; D003872:Dermatitis / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D004817:Epidermis / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D005260:Female; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D015603:Keratinocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008156:Luciferases; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D009504:Neutrophils / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D053832:Surgical Tape / Q000009:adverse effects*
chemical_list: D018925:Chemokines; D012333:RNA, Messenger; D000255:Adenosine Triphosphate; D008156:Luciferases; D001081:Apyrase