title: In Vivo Imaging of Retinal Hypoxia in a Model of Oxygen-Induced Retinopathy.
abstract: Ischemia-induced hypoxia elicits retinal neovascularization and is a major component of several blinding retinopathies such as retinopathy of prematurity (ROP), diabetic retinopathy (DR) and retinal vein occlusion (RVO). Currently, noninvasive imaging techniques capable of detecting and monitoring retinal hypoxia in living systems do not exist. Such techniques would greatly clarify the role of hypoxia in experimental and human retinal neovascular pathogenesis. In this study, we developed and characterized HYPOX-4, a fluorescence-imaging probe capable of detecting retinal-hypoxia in living animals. HYPOX-4 dependent in vivo and ex vivo imaging of hypoxia was tested in a mouse model of oxygen-induced retinopathy (OIR). Predicted patterns of retinal hypoxia were imaged by HYPOX-4 dependent fluorescence activity in this animal model. In retinal cells and mouse retinal tissue, pimonidazole-adduct immunostaining confirmed the hypoxia selectivity of HYPOX-4. HYPOX-4 had no effect on retinal cell proliferation as indicated by BrdU assay and exhibited no acute toxicity in retinal tissue as indicated by TUNEL assay and electroretinography (ERG) analysis. Therefore, HYPOX-4 could potentially serve as the basis for in vivo fluorescence-based hypoxia-imaging techniques, providing a tool for investigators to understand the pathogenesis of ischemic retinopathies and for physicians to address unmet clinical needs.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Md Imam
authors.initials: MI
authors.lastname: Uddin
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Evans
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Jason R
authors.initials: JR
authors.lastname: Craft
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Megan E
authors.initials: ME
authors.lastname: Capozzi
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Gary W
authors.initials: GW
authors.lastname: McCollum
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Rong
authors.initials: R
authors.lastname: Yang
authors.affiliation: A. B. Hancock, Jr., Memorial Laboratory for Cancer Research, Departments of Biochemistry, Chemistry and Pharmacology, Vanderbilt Institute of Chemical Biology, Center for Molecular Toxicology and Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Marnett
authors.affiliation: A. B. Hancock, Jr., Memorial Laboratory for Cancer Research, Departments of Biochemistry, Chemistry and Pharmacology, Vanderbilt Institute of Chemical Biology, Center for Molecular Toxicology and Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Md Jashim
authors.initials: MJ
authors.lastname: Uddin
authors.affiliation: Pharma Research and Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche, Ltd., Basel, Switzerland.
authors.forename: Ashwath
authors.initials: A
authors.lastname: Jayagopal
authors.affiliation: Department of Ophthalmology and Visual Sciences, Vanderbilt University School of Medicine, Nashville, TN, USA.|Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: John S
authors.initials: JS
authors.lastname: Penn
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005456:Fluorescent Dyes / Q000008:administration & dosage; D000860:Hypoxia / Q000000981:diagnostic imaging* / Q000473:pathology*; D000069416:Intravital Microscopy / Q000379:methods; D051379:Mice; D009389:Neovascularization, Pathologic / Q000000981:diagnostic imaging; D061848:Optical Imaging / Q000379:methods*; D012160:Retina / Q000000981:diagnostic imaging* / Q000473:pathology*; D012164:Retinal Diseases / Q000000981:diagnostic imaging* / Q000473:pathology*
chemical_list: D005456:Fluorescent Dyes