title: The electrochemical and fluorescence detection of nitric oxide in the cochlea and its increase following loud sound.
abstract: A nitric oxide (NO)-selective sensor (tip diameter 30 microm) was inserted into the perilymph of the basal turn of the guinea pig cochlea. The basal level and stimulation-induced changes of NO were measured. The mean (+/-S.E.M.) basal level of NO was 273+/-42.9 nM. Following perilymphatic perfusion of the artificial perilymph containing NO synthase (NOS) substrate L-arginine (100 microM) combined with cofactor (6R)-5,6,7,8-tetrahydrobiopterin dihydrochloride (100 microM), a rapid and significant increase of NO to a mean concentration of 392+/-32.3 nM (P < 0.01, n = 10) was recorded. In contrast, a significant decrease of mean NO concentration to 180+/-32.7 nM (P < 0.01, n = 10) was observed following the perfusion of the NOS-inhibiting agent N(G)-nitro-L-arginine methyl ester (100 microM). No change in the NO concentration was found following the perfusion of either artificial perilymph or N(G)-monomethyl-D-arginine (100 microM) solution employed as controls. Broadband noise exposure (3 h/day at 120 dBA SPL) for three consecutive days produced an increase in NO concentration to 618+/-60.7 nM (P < 0.05, n = 10) in the perilymph. In addition, by using specific dyes for NO, 4,5-diaminofluoresceine diacetate and for the reactive oxygen species (ROS), dihydrorhodamine 1,2,3, the distribution of NO in the whole mounts of the organ of Corti and the production of ROS in vivo in the organ of Corti were investigated in both control (n = 5) and noise-exposed (n = 5) animals. The more intense NO and ROS fluorescence was observed in both the inner and outer hair cells in the noise-exposed groups. It is proposed that both the basal level and the increase in NO concentration following the addition of substrate (L-arginine) are produced by the constitutive NOS while the elevated NO and ROS following noise exposure indicate that NO may be involved in noise-induced hearing loss.
authors.affiliation: Department of Otolaryngology, the General Hospital of the CPAPA, Beijing 100039, PR China.
authors.forename: Xiaorui
authors.initials: X
authors.lastname: Shi
authors.forename: Tianying
authors.initials: T
authors.lastname: Ren
authors.forename: Alfred L
authors.initials: AL
authors.lastname: Nuttall
mesh_terms: D000818:Animals; D003051:Cochlea / Q000293:injuries* / Q000378:metabolism*; D004563:Electrochemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016057:Evoked Potentials, Auditory, Brain Stem; D005260:Female; D005453:Fluorescence; D006168:Guinea Pigs; D006317:Hearing Loss, Noise-Induced / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D008954:Models, Biological; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009622:Noise / Q000009:adverse effects*; D009925:Organ of Corti / Q000293:injuries / Q000378:metabolism; D010498:Perilymph / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester