Endothelial nitric oxide synthase upregulation in the cochlea of the guinea pig after intratympanic gentamicin injection

Abstract

Single-shot transtympanic gentamicin therapy has become a popular treatment modality for Meniere's disease despite the known possible ototoxic properties of this drug. It was shown recently that NO production and iNOS were upregulated after gentamicin application, which was interpreted as a possible effect of ototoxicity. In this study we analyzed the expression of eNOS after gentamicin application to determine a possible correlation of this enzyme with gentamicin-induced ototoxicity. We compared eNOS expression in gentamicin-treated and non-treated guinea pigs in the second turn of the cochlea, an area corresponding to speech perception in humans. Gentamicin (4 mg) was injected intratympanically into the middle ear of guinea pigs ( n =3) and the reduction of the hearing threshold level was determined by recording acoustic-evoked potentials (AEP) before and 5 days after gentamicin application. Morphological alterations in the organ of Corti were analyzed by light and electron microscopy. Gold-labeled anti-eNOS antibodies were counted in eight different cell areas for quantification of eNOS expression. Seven animals were analyzed as controls. After gentamicin application, a deterioration of hearing level was observed varying from 10 to 30 dB. A high degree of vacuolization was identified in the third row of outer hair cells. At the subcellular level, the subsurface cisterns in outer hair cells were dissociated from the basolateral cell membrane, and the mitochondrial membranes were frequently damaged. Statistically significant upregulation of eNOS was observed in all cell types analyzed. Depending on the various cell types the amount of gold-labeled eNOS antibodies was 2.5 to 5.7 times higher after gentamicin application. We observed significant eNOS upregulation after gentamicin application in the cochlea, in conjunction with cellular damages and decreased hearing.