The effect of explantation and neomycin on hair cells and supporting cells in organotypic cultures of the adult guinea-pig utricle

Abstract

Recent reports suggest that immature hair bundles are observed following aminoglycoside-induced hair-cell loss in the mammalian utricle in vitro as well as in vivo. It is therefore important to document the initial morphological changes associated with both culturing and aminoglycoside application so that degeneration can be clearly distinguished from regeneration. In this study, utricles from adult guinea pigs were maintained in culture for either 3 or 8 days, half being exposed to neomycin for days 2 and 3. They were then processed for microscopical examination and compared with control utricles from animals of the same age. The numbers of hair-cell and supporting-cell nuclei were counted and hair-cell morphology assessed. Bundles were classified as having either stepped (SHB) or unstepped (UHB) stereocilia, and their density determined. The numbers of hair-cell, but not supporting-cell, nuclei declined significantly compared with controls in both untreated and treated explants, the greatest reduction occurring 5 days after neomycin administration. The density of SHBs also declined but there was no significant change in UHB density, resulting in a residual population of hair bundles of more immature appearance in both untreated and treated utricles in vitro than in vivo. Although degenerative events such as hair-cell ejection from, or retraction into, the sensory epithelium were observed, no evidence of regeneration was found.