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Ouabain is a common tool to explore the pathophysiological changes in adult mammalian cochlea  in vivo . In prior studies, locally administering ouabain via round window membrane demonstrated that the ototoxic effects of ouabain  in vivo  varied among mammalian species. Little is known about the ototoxic effects  in vitro . Thus, we prepared cochlear organotypic cultures from postnatal day-3 rats and treated these cultures with ouabain at 50, 500, and 1000   μ  M for different time to elucidate the ototoxic effects of ouabain  in vitro  and to provide insights that could explain the comparative ototoxic effects of ouabain  in vivo . Degeneration of cochlear hair cells and spiral ganglion neurons was evaluated by hair-cell staining and neurofilament labeling, respectively. Annexin V staining was used to detect apoptotic cells. A quantitative RT-PCR apoptosis-focused gene array determined changes in apoptosis-related genes. The results showed that ouabain-induced damage  in vitro  was dose and time dependent. 500   μ  M ouabain and 1000   μ  M ouabain were destructively traumatic to both spiral ganglion neurons and cochlear hair cells in an apoptotic signal-dependent pathway. The major apoptotic pathways in ouabain-induced spiral ganglion neuron apoptosis culminated in the stimulation of the p53 pathway and triggering of apoptosis by a network of proapoptotic signaling pathways.

Ouabain-Induced Apoptosis in Cochlear Hair Cells and Spiral Ganglion NeuronsIn Vitro