[Streptomycin-induced apoptosis of rat cochlear hair cell cultured in vitro].
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.
He
Jing-chun
JC
Department of Otorhinolaryngology, Affiliated Sixth People's Hospital, Shanghai Jiaotong University, Otolaryngology Institute of Shanghai Jiaotong University, Shanghai 200233, China.
Yu
Dong-zhen
DZ
Ding
Da-lian
DL
Yin
Shan-kai
SK
Salvi
Richard J
RJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053178:Caspase 6 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D006198:Hair Cells, Auditory / Q000166:cytology / Q000187:drug effects*; D051381:Rats; D011916:Rats, Inbred F344; D013307:Streptomycin / Q000009:adverse effects*
C505518:Casp6 protein, rat; C505528:Casp8 protein, rat; C506435:Casp9 protein, rat; D053178:Caspase 6; D053181:Caspase 8; D053453:Caspase 9; D013307:Streptomycin