Noise-induced nitrotyrosine increase and outer hair cell death in guinea pig cochlea.
BACKGROUND
Modern research has provided new insights into the biological mechanisms of noise-induced hearing loss, and a number of studies showed the appearance of increased reactive oxygen species (ROS) and reactive nitrogen species (RNS) during and after noise exposure. This study was designed to investigate the noise exposure induced nitrotyrosine change and the mechanism of outer hair cells death in guinea pig cochlea.


METHODS
Thirty guinea pigs were used in this study. The experimental animals were either exposed for 4 hours per day to broadband noise at 122 dB SPL (A-weighted) for 2 consecutive days or perfused cochleae with 5 mg/ml of the SIN1 solutions, an exogenous NO and superoxide donor, for 30 minutes. Then the cochleae of the animals were dissected. Propidium iodide (PI), a DNA intercalating fluorescent probe, was used to trace morphological changes in OHC nuclei. The distribution of nitrotyrosine (NT) in the organ of Corti and the cochlear lateral wall tissue from the guinea pigs were examined using fluorescence immunohistochemistry method. Whole mounts of organ of Corti were prepared. Morphological and fluorescent changes were examined under a confocal microscope.


RESULTS
Either after noise exposure or after SIN1 perfusion, outer hair cells (OHCs) death with characteristics of both apoptotic and necrotic degradation appeared. Nitrotyrosine immunolabeling could be observed in the OHCs from the control animals. After noise exposure, NT immunostaining became much greater than the control animals in OHCs. The apoptotic OHC has significant increase of nitrotyrosine in and around the nucleus following noise exposure. In the normal later wall of cochleae, relatively weak nitrotyrosine immunolabeling could be observed. After noise exposure, nitrotyrosine immunoactivity became stronger in stria vascularis.


CONCLUSIONS
Noise exposure induced increase of nitrotyrosine production is associated with OHCs death suggesting reactive nitrogen species participation in the cochlear pathophysiology of noise-induced hearing loss.
Department of Otolaryngology Head and Neck Surgery, Chinese People's Liberation Army General Hospital, Beijing 100853, China. hanweiju@aliyun.com
Wei-ju
WJ
Han
Xiao-rui
XR
Shi
Alfred
A
Nuttall
D000818:Animals; D016923:Cell Death; D003051:Cochlea / Q000737:chemistry / Q000473:pathology*; D005260:Female; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000473:pathology*; D007150:Immunohistochemistry; D008297:Male; D009622:Noise / Q000009:adverse effects*; D009925:Organ of Corti / Q000737:chemistry / Q000473:pathology; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
C002744:3-nitrotyrosine; D014443:Tyrosine