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[Streptomycin-induced apoptosis of rat cochlear hair cell cultured in vitro]
Jing-chun He1, Dong-zhen Yu, Da-lian Ding
1Department of Otorhinolaryngology, Affiliated Sixth People's Hospital, Shanghai Jiaotong University, Otolaryngology Institute of Shanghai Jiaotong University, Shanghai 200233, China.
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 mmol/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.
