Topographic and quantitative evaluation of gentamicin-induced damage to peripheral innervation of mouse cochleae

Abstract

Ototoxicity induced by aminoglycoside antibiotics appears to occur both in hair cells (HCs) and the cochlear nerves that innervate them. Although HC loss can be easily quantified, neuronal lesions are difficult to quantify because two types of afferent dendrites and two types of efferent axons are tangled beneath the hair cells. In the present study, ototoxicity was induced by gentamicin in combination with the diuretic agent furosemide. Neuronal lesions were quantified in cochlear whole-mount preparations combined with microsections across the habenular perforate (HP) openings to achieve a clear picture of the topographic relationship between neuronal damage and HC loss. Multiple immunostaining methods were employed to differentiate the two types of afferent dendrites and two types of efferent axons. The results show that co-administration of gentamicin and furosemide resulted in a typical dynamic pattern of HC loss that spread from the basal turn to the outer hair cells to the apex and inner hair cells, depending on the dose and survival time after drug administration. Lesions of the innervation appeared to occur at two stages. At the early stage (2-4 days), the loss of labeling of the two types of afferent dendrites was more obvious than the loss of labeled efferent axons. At the late stage (2-4 weeks), the loss of labeled efferent axons was more rapid. In the high-dose gentamicin group, the loss of outer HCs was congruent with afferent dendrite loss at the early stage and efferent axon loss at the late stage. In the low-dose gentamicin group, the loss of labeling for cochlear innervation was more severe and widespread. Thus, we hypothesize that the gentamicin-induced damage to cochlear innervation occurs independently of hair cell loss.

Keywords: 4′,6-diamidino-2-phenylindole; A; Afferent dendrites; B; DAPI; Efferent axons; HC; HP; Hair cells; IHC; LOC; M; MOC; Mouse; OHC; OSL; SGNs; Spiral ganglion neurons; TMRD; Tetramethyl rhodamine dextran; apex segment; basal segment; habenular perforate; hair cell; inner hair cell; lateral olivary complex; medial olivary complex; middle segment; osseous spiral lamina; outer hair cell; spiral ganglion neurons; tetramethylrhodamine-conjugated dextran.