Figure 7
Modulation of gentamicin-conjugated Texas Red (GTTR) uptake and hair cell survival following exposure to calcium ions. Cochlear explants were pretreated with Ca2+ (1 or 2 mM) for 10 min. (a) Cochlear explants were incubated with GTTR (500 μM) for 30 min in the absence and presence of Ca2+ (1 or 2 mM). The samples were washed and fixed in 4% paraformaldehyde (PFA) and stained with fluorescein isothiocyanate (FITC)-labeled palloidin for 30 min. The specimens were observed under a fluorescent microscope. (b) Cochlear explants were incubated with 300 μM gentamicin for 24 h in the absence and presence of Ca2+ (1 or 2 mM). After fixation, the specimens were stained with phalloidin–tetramethylrhodamine isothiocyanate (TRITC) and examined under a fluorescent microscope. (c) Cochlear explants were incubated with or without Ca2+ (1 or 2 mM) for 12 h. Cochlear explants treated with various Ca2+ concentrations were protected against gentamicin. Total cell lysates from the organ of Corti were subjected to 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with transient receptor potential vanilloid 1 (TRPV1) and TRPV4 antibodies.
