title: Functional hair cell mechanotransducer channels are required for aminoglycoside ototoxicity.
abstract: Aminoglycosides (AG) are commonly prescribed antibiotics with potent bactericidal activities. One main side effect is permanent sensorineural hearing loss, induced by selective inner ear sensory hair cell death. Much work has focused on AG's initiating cell death processes, however, fewer studies exist defining mechanisms of AG uptake by hair cells. The current study investigated two proposed mechanisms of AG transport in mammalian hair cells: mechanotransducer (MET) channels and endocytosis. To study these two mechanisms, rat cochlear explants were cultured as whole organs in gentamicin-containing media. Two-photon imaging of Texas Red conjugated gentamicin (GTTR) uptake into live hair cells was rapid and selective. Hypocalcemia, which increases the open probability of MET channels, increased AG entry into hair cells. Three blockers of MET channels (curare, quinine, and amiloride) significantly reduced GTTR uptake, whereas the endocytosis inhibitor concanavalin A did not. Dynosore quenched the fluorescence of GTTR and could not be tested. Pharmacologic blockade of MET channels with curare or quinine, but not concanavalin A or dynosore, prevented hair cell loss when challenged with gentamicin for up to 96 hours. Taken together, data indicate that the patency of MET channels mediated AG entry into hair cells and its toxicity. Results suggest that limiting permeation of AGs through MET channel or preventing their entry into endolymph are potential therapeutic targets for preventing hair cell death and hearing loss.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Stanford University, Stanford, California, United States of America.
authors.forename: Abdelrahman
authors.initials: A
authors.lastname: Alharazneh
authors.forename: Lauren
authors.initials: L
authors.lastname: Luk
authors.forename: Markus
authors.initials: M
authors.lastname: Huth
authors.forename: Ashkan
authors.initials: A
authors.lastname: Monfared
authors.forename: Peter S
authors.initials: PS
authors.lastname: Steyger
authors.forename: Alan G
authors.initials: AG
authors.lastname: Cheng
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Ricci
mesh_terms: D000617:Aminoglycosides / Q000633:toxicity*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects; D004705:Endocytosis / Q000187:drug effects; D005839:Gentamicins / Q000633:toxicity; D006198:Hair Cells, Auditory / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D021621:Imaging, Three-Dimensional; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000378:metabolism*; D007700:Kinetics; D040542:Mechanotransduction, Cellular / Q000187:drug effects*; D051379:Mice; D017785:Photons; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014966:Xanthenes / Q000378:metabolism
chemical_list: D000617:Aminoglycosides; D005839:Gentamicins; D007473:Ion Channels; D014966:Xanthenes; C034657:Texas red