Sound damage and gentamicin treatment produce different patterns of damage to the efferent innervation of the chick cochlea.

Both sound exposure and gentamicin treatment cause damage to sensory hair cells in the peripheral chick auditory organ, the basilar papilla. This induces a regeneration response which replaces hair cells and restores auditory function. Since functional recovery requires the re-establishment of connections between regenerated hair cells and the central nervous system, we have investigated the effects of sound damage and gentamicin treatment on the neuronal elements within the cochlea. Whole-mount preparations of basilar papillae were labeled with phalloidin to label the actin cytoskeleton and antibodies to neurofilaments, choline acetyltransferase, and synapsin to label neurons; and examined by confocal laser scanning microscopy. When chicks are treated with gentamicin or exposed to acoustic overstimulation, the transverse nerve fibers show no changes from normal cochleae assayed in parallel. Efferent nerve terminals, however, disappear from areas depleted of hair cells following acoustic trauma. In contrast, efferent nerve endings are still present in the areas of hair cell loss following gentamicin treatment, although their morphological appearance is greatly altered. These differences in the response of efferent nerve terminals to sound exposure versus gentamicin treatment may account, at least in part, for the discrepancies reported in the time of recovery of auditory function.

Distribution of nerve fibers in the basilar papilla of normal and sound-damaged chick cochleae.

Epifluorescent light microscopy and confocal laser scanning microscopy were employed to visualize the distribution of nerve fibers in whole-mount preparations of normal and sound-damaged chick basilar papillae (BP). In normal cochleae, we identified a consistent pattern of nerve processes that ran transversely across the BP. The transverse processes increase in number from the proximal to the distal ends of the epithelium. However, when the processes are separated into populations of thin fibers and thick bundles, the thin fibers are more prevalent in distal regions whereas thick bundles are more extensive in proximal regions. Furthermore, the thick bundles form an elaborate longitudinal network in the border cell and hyaline cell region. Based on these data and no other previous studies, the thin fibers appear to be afferent nerves and the thick bundles represent efferent nerves. When birds are exposed to acoustic trauma, the normal pattern and number of nerve processes is not altered by levels of sound that produce moderate levels of damage, i.e., damage that leads to hair cell loss and regeneration. However, the nerve pattern is disrupted by severe levels of damage that destroy both hair cells and supporting cells. These findings indicate that the level of sound exposure that induces hair cell regeneration may damage the synaptic endings associated with the lost hair cells, but that the nerve processes that give rise to these endings remain intact within the sensory epithelium. In contrast, severe damage destroys both the hair cells and their associated nerve fibers.

Migration of hyaline cells into the chick basilar papilla during severe noise damage.

Severe acoustic damage in the chick cochlea causes a destruction of both hair cells and supporting cells in a localized area on the basilar papilla. In this region, the sensory cells are replaced by a layer of flattened epithelial cells. We have employed scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) to examine the structure and cytoskeletal changes involved in this process. Immunocytochemical staining for actin indicates that the flattened cells are derived from the hyaline cells normally located along the inferior edge of the basilar papilla. In control cochleae the hyaline cells contain dense bundles of actin filaments that anchor into the basal surface of the cells. The hyaline cells appear to redistribute into the severely damaged region by extending the actin bundles at their basal surfaces. Moreover, the efferent nerves that normally form a network among the hyaline cells move into the severely damaged area along with the hyaline cells. In moderately damaged cochleae, where only hair cells are lost, the hyaline cells do not spread into the damaged region. The functional role of this hyaline cell migration is unknown, but it may be involved in maintenance or repair of the severely damaged cochlea.