OBJECTIVETo investigate the upstaging and accumulation of gentamicin by mouse hair cells in vitro.

METHODSCochlear explants were prepared from the microdissected neonatal mouse cochlea. Cochlear explants were cultured with gentamicin-Texas-red conjunction (GTTR) for different time. Laser confocal microscopy was used to observe the distribution of GTTR in the cochlear sensory cells after labeling with phalloidin-alexa-488.

RESULTSSoon after culture, there was diffuse red staining all tissue cells in the explants. At later time the hair cells were more staining than other cells in the explants. There was no obviously accumulation of GTTR in the supporting cells. The peak level of fluorescent density was reached at 24 hours culture. The GTTR was seen in the infracuticular zone of the hair cells. There was still accumulation of GTTR in the hair cells of the explants after 7 days culturing.

CONCLUSIONSGTTR and cochlea explants were useful methods to investigate the pharmacokinetics and mechanisms of gentamicin accumulation over time.