title: Intravital analysis of vascular permeability in mice using two-photon microscopy.
abstract: Blood vessel endothelium forms a semi-permeable barrier and its permeability controls the traffics of plasma contents. Here we report an intravital evaluation system for vascular permeability in mice using two-photon microscopy. We used various sizes of fluorescein-conjugated dextran as a tracer and its efflux was quantified by measuring the changes of fluorescent intensity both on the blood vessel area and the interstitial space. Using this system, we demonstrated that skin blood vessels limited the passage of dextran larger than 70 kDa under homeostatic conditions. We evaluated the kinetics of vascular permeability in histamine- or IgE-induced type I allergic models and a hapten-induced type IV allergic model. In such inflammatory conditions, the hyperpermeability was selectively induced in the postcapillary venules and dextran as large as 2000-kDa leaked from the bloods. Taken together, our study provides a convenient method to characterize the skin blood vessels as a traffic barrier in physiological conditions.
authors.affiliation: Department of Dermatology, Kyoto University Graduate School of Medicine, Kyoto, Japan.
authors.forename: Gyohei
authors.initials: G
authors.lastname: Egawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nakamizo
authors.forename: Yohei
authors.initials: Y
authors.lastname: Natsuaki
authors.forename: Hiromi
authors.initials: H
authors.lastname: Doi
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Miyachi
authors.forename: Kenji
authors.initials: K
authors.lastname: Kabashima
mesh_terms: D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D003911:Dextrans / Q000008:administration & dosage; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006632:Histamine / Q000633:toxicity; D006967:Hypersensitivity / Q000209:etiology / Q000473:pathology*; D007073:Immunoglobulin E / Q000633:toxicity; D007249:Inflammation / Q000139:chemically induced / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D017785:Photons; D012867:Skin / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology
chemical_list: D003911:Dextrans; D007073:Immunoglobulin E; D006632:Histamine; D016650:Fluorescein-5-isothiocyanate