Streptomycin-induced apoptosis of rat cochlear hair cell cultured in vitro
10.3760/cma.j.issn.1673-0860.2009.06.012
- VernacularTitle:硫酸链霉素引起的大鼠体外培养耳蜗毛细胞凋亡
- Author:
Jing-Chan HE
1
;
Dong-Zhen YU
;
Da-Lian DING
;
Shan-Kai YIN
;
Salvi J RICHARD
Author Information
1. 上海交通大学医学院附属第六人民医院
- Keywords:
Hair cells,auditory;
Organ culture techniques;
Streptomycin;
Apoptosis;
Caspases
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2009;44(6):494-498
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate if caspase pathway was involved in streptomycin-induced cell apoptosis in cochlear hair cells. Methods F344 rats at postnatal day 3 or 4 were used for the study in cochlear organotypic cultures. The cochlear basilar membrane was micro-dissected out and cultured overnight, and then treated with 1 mmol/L streptomycin for 24 hours. Before the termination, the activity of caspase-8, 9 or 6 were detected with FAM-peptide-FMK labeled caspase-8, 9 or 6, respectively. The stereocilia and cuticular plate of hair cells were stained with TRITC conjugated phalloidin, and the nuclei were stained with Topro-3 DNA probe. The specimens were observed and photographed under confocal fluorescent microscope. Results Streptomycin with 1 nunol/L causes about 80% cochlear hair cells missing in the basal turn and 10% hair cell loss in the apex. After streptomycin treatment, the nuclear shrinkage and fragmentation were found in most cochlear hair cells, and the caspase-8, caspase-9 and caspase-6 were greatly activated. Conclusions Apoptosis is involved in the cochlear hair cells death induced by Streptomycin in vitro. The caspase activities in upstream and downstream are maybe the major apoptotic pathway.
