Rapid fluorescent vital imaging of olfactory epithelium
Hironobu Nishijima,
Matthew J. Zunitch,
Masafumi Yoshida,
Kenji Kondo,
Tatsuya Yamasoba,
James E. Schwob,
Eric H. Holbrook
Affiliations
Hironobu Nishijima
Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA; Department of Otolaryngology Head and Neck Surgery, Harvard Medical School, Massachusetts Eye and Ear, 243 Charles Street, Boston, MA 02114, USA; Department of Otolaryngology-Head and Neck Surgery, The University of Tokyo, Tokyo 113-0033, Japan
Matthew J. Zunitch
Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA
Masafumi Yoshida
Department of Otolaryngology-Head and Neck Surgery, The University of Tokyo, Tokyo 113-0033, Japan
Kenji Kondo
Department of Otolaryngology-Head and Neck Surgery, The University of Tokyo, Tokyo 113-0033, Japan
Tatsuya Yamasoba
Department of Otolaryngology-Head and Neck Surgery, The University of Tokyo, Tokyo 113-0033, Japan
James E. Schwob
Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA
Eric H. Holbrook
Department of Developmental, Molecular, and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA; Department of Otolaryngology Head and Neck Surgery, Harvard Medical School, Massachusetts Eye and Ear, 243 Charles Street, Boston, MA 02114, USA; Corresponding author
Summary: Olfactory epithelium (OE) undergoes degeneration in disorders such as age-related and post-viral olfactory dysfunction. However, methods for real-time in vivo detection of OE and assessment of total extent within the nasal cavity are currently unavailable. We identified two fluorescence probes for rapidly detecting and evaluating the entire extent of mice OE with topical application. Taking advantage of the differential expression of the enzymes cytochrome p450 (CYP) and γ-glutamyltranspeptidase (GGT) in OE relative to respiratory epithelium, we utilized the conversion of coumarin (a substrate of various CYP subtypes) and gGlu-HRMG (a substrate of GGT) by these enzymes to form metabolites with fluorescent emissions in the duct cells and sustentacular cells of neuron-containing OE. In depleted and regenerated OE model, the emission of these probes remained absent in respiratory metaplasia but appeared in regenerated OE. These substrates could be used to monitor OE degeneration and follow regenerative response to therapeutic interventions.
