Aby wyświetlić tę treść, wymagana jest subskrypcja JoVE. Zaloguj się lub rozpocznij bezpłatny okres próbny.
Here, we present a protocol for a mouse model of noise-induced hearing loss (NIHL). To induce NIHL, we developed a new and simple device using corrugated plastic, a rat trap cage, and a speaker. Auditory brainstem response and immunofluorescence imaging were employed to assess the hearing function and outer hair cell damage, respectively.
An animal model of noise-induced hearing loss (NIHL) is useful for pathologists, therapists, pharmacologists, and hearing researchers to thoroughly understand the mechanism of NIHL, and subsequently optimize the corresponding treatment strategies. This study aims to create an improved protocol for developing a mouse model of NIHL. Male C57BL/6J mice were used in this study. Unanesthetized mice were exposed to loud noises (1 and 6 kHz, presented simultaneously at 115-125 dB SPL-A) continuously for 6 h per day for 5 consecutive days. Auditory function was assessed 1 day and 1 week after noise exposure, using auditory brainstem response (ABR). After the ABR measurement, the mice were sacrificed, and their organs of Corti were collected for immunofluorescence staining. From the auditory brainstem response (ABR) measurements, significant hearing loss was observed 1 day after noise exposure. After 1 week, the hearing thresholds of the experimental mice decreased to ~80 dB SPL, which was still a significantly higher level than the control mice (~40 dB SPL). From the results of immunofluorescence imaging, outer hair cells (OHCs) were shown to be damaged. In summary, we created a model of NIHL using male C57BL/6J mice. A new and simple device for generating and delivering pure-tone noise was developed and then employed. Quantitative measurements of hearing thresholds and morphological confirmation of OHC damage both demonstrated that the applied noise successfully induced an expected hearing loss.
About 1.3 billion people worldwide suffer from hearing loss due to noise exposure1. In this study, we aimed to establish a clear step-by-step process for inducing and confirming noise-induced hearing loss (NIHL). NIHL results from a degeneration/destruction of the hair cells (HCs) and spiral ganglion neurons (SGNs), damage in the HC stereocilia, and/or loss of synapses between the cochlear inner HCs and SGNs. Such abnormalities may also cause tinnitus and impaired speech perception (especially in a complex acoustic environment) besides NIHL. Social, psychological, and cognitive functions may be sequentially affected by these physiological defic....
Animal experiments in this study were approved by the Animal Care Committee of Mackay Medical College. Eight-week-old Male C57BL/6J mice were purchased from the National Laboratory Animal Center (New Taipei City, Taiwan). All mice were bred and housed in accordance with the standard animal protocol.
1. Induction of NIHL in mice
A shift in ABR hearing threshold
The hearing threshold of the mice was measured using tone-burst ABR either 1 day or 1 week after the noise exposure. A significant increase in the hearing threshold at all three tested frequencies was observed (12 kHz: 84.29 ± 2.77 dB SPL; 24 kHz: 91.43 ± 0.92 dB SPL; 32 kHz: 98.57 ± 1.43 dB SPL) 1 day after the noise exposure (i.e., the 6th day). Partial hearing recovery occurred 1 week after the noise exposure (i.e., the 13th d.......
NIHL can be divided into two types: temporary NIHL, which shows a temporal shift of the hearing threshold, and permanent NIHL, which is featured by a permanent hearing-threshold shift. The hearing loss that we observed on the 6th day (1 day after the noise exposure) is believed to be a combination of these two types. In this case, the hearing threshold would show a gradual recovery over time owing to the temporal component of hearing loss. In our preliminary experimental studies, the results acquired with the .......
No conflict of interest to disclose.
We thank the grants from the Ministry of Science and Technology (MOST) of the Taiwan Government (MOST 110-2314-B-715-005, MOST 111-2314-B-715-009-MY3), and intramural research grants from Mackay Medical College (MMC-RD-110-1B-P030, MMC-RD-111-CF-G002-03).
....Name | Company | Catalog Number | Comments |
1/4" CCP Free-field Standard Microphone Set | GRAS | 428158 | For noise exposure |
Amplifier Input Module, AMI100D | BIOPAC | For auditory brainstem response | |
Bio-amplifier, BIO100C | BIOPAC | For auditory brainstem response | |
Bovine Serum Albumin | SIGMA | A9647 | Immunofluorescence staining |
Cellsens software | Olympus life science | Image acquisition | |
Corrugated plastic | |||
DAPI fluoromount | SouthernBiotech | 0100-20 | Immunofluorescence staining |
Ethylenediaminetetraacetic acid | SIGMA | E5134 | Decalcification |
Evoked Response Amplifier, ERS100C | BIOPAC | For auditory brainstem response | |
Formaldehyde | APLHA | F030410 | Fixation of cochlear |
High Performance Data Acquisition System, MP160 | BIOPAC | For auditory brainstem response | |
Modular Extension Cable, MEC110C | BIOPAC | For auditory brainstem response | |
Myo7A primary antibody | Proteus | 25-6790 | Immunofluorescence staining |
Myo7A secondary antibody | Jackson immunoresearch | 711-545-152 | Immunofluorescence staining |
Needle Electrode, Unipolar 12 mmTp, EL452 | BIOPAC | For auditory brainstem response | |
phalloidin antibody | Alexa Fluor | A12381 | Immunofluorescence staining |
phosphate-buffered saline | SIGMA | P4417 | |
Rat trap cage | 14 cm x 17 cm x 24cm | ||
ROMPUN- xylazine injection, solution | Bayer HealthCare, LLC | ||
Sound amplifier, MT-1000 | unika | For noise exposure | |
Sound generator/analyzer/miscellaneous, FW-02 | CLIO | 620300719 | For noise exposure |
Soundproof chamber | IEA Electro-Acoustic Technology | For noise exposure and ABR | |
Speaker | IEA Electro-Acoustic Technology | For noise exposure | |
Stimulator Module, STM100C | BIOPAC | For auditory brainstem response | |
Triton X-100 | SIGMA | T8787 | Immunofluorescence staining |
Tubephone Set, OUT101 | BIOPAC | For auditory brainstem response | |
Upright Microscope, BX53 | Olympus | Image acquisition | |
Zoletil | Virbac |
Zapytaj o uprawnienia na użycie tekstu lub obrazów z tego artykułu JoVE
Zapytaj o uprawnieniaThis article has been published
Video Coming Soon
Copyright © 2025 MyJoVE Corporation. Wszelkie prawa zastrzeżone