A BAD link to mitochondrial cell death in the cochlea of mice with noise-induced hearing loss.

Acoustic overstimulation induces calcium overload and activation of mitochondria-mediated cell death pathways in outer hair cells (OHC) of the cochlea. However, it is not known whether these events are interrelated or independent. We have recently reported that the calcium-dependent phosphatase calcineurin is activated in OHC following noise exposure and now postulate that calcium overload triggers mitochondria-mediated death pathways through activation of Bcl-2-associated death promoter (BAD) by calcineurin. CBA/J mice were exposed to broadband noise (2-20 kHz), causing a permanent threshold shift of about 40 dB at 12 and 20 kHz, corresponding to damage in the middle and basal turns of the cochlea. Loss of OHC in the basal region was evident in surface preparations. BAD immunostaining in control animals had a cytoplasmic distribution in the cells of the organ of Corti. Five hours after acoustic overstimulation, mitochondria and BAD redistributed to the perinuclear region of OHC in the basal and middle turns but not in the apical turn. The nonapoptotic phospho-BAD (Ser 112) was up-regulated in cells undamaged by noise (supporting cells and inner hair cells) but not in OHC. These data establish a connection between calcium overload and mitochondria-mediated death pathways in OHC and also suggest a dual role for BAD. The translocation of BAD to the mitochondria in degenerating cells is indicative of the activation of its proapoptotic capacity, whereas up-regulation of phospho-BAD is consistent with a nonapoptotic role of BAD in less vulnerable cells.

Biochemical evidence for the presence of serotonin transporters in the rat cochlea.

Cochlear serotonergic innervation is constituted by efferent fibers projecting both to the area below the inner and the outer hair cells. Previous detection of serotonin (5-HT) metabolites and 5-HT receptor mRNAs suggests the existence of serotonergic synaptic activity in the cochlea. The present study explores this possibility through the effect of 6-nitroquipazine (6-NQ), a 5-HT selective reuptake inhibitor, on the basal turnover of 5-HT. The concentrations of 5-HT and its metabolite 5-hydroxyindole-3-acetic acid (5-HIAA) were quantified by high performance liquid chromatography with electrochemical detection in blood-free cochleae of rats treated with 6-NQ or saline and kept under silent conditions. Treatment with 6-NQ induced a significant increase of the cochlear concentration of 5-HT and a significant reduction of 5-HIAA concentration with respect to saline treatment. These findings could indicate that 6-NQ induced the blockade of the 5-HT selective reuptake to the cochlear serotonergic fibers. This suggests that plasma membrane 5-HT transporters are present in cochlear serotonergic fibers. Even though the role of serotonergic innervation on cochlear physiology remains unknown, the existence of cochlear serotonergic synaptic activity is strongly supported by present contributions.

Effects of aging on cochlear monoamine turnover.

The aging of the cochlear dopaminergic system has been analyzed by quantifying the levels of dopamine (DA) and its metabolites (3,4-dihydroxyphenylacetic acid, DOPAC, and homovanillic acid, HVA) in adult rats aged 3, 12 or 24 months. The main results were an increase in DA, DOPAC and HVA basal concentrations in aged females with respect to the adults (3 or 12 months old), while just DA and DOPAC increased in aged males. A higher synthesis of DA in aged animals could support these findings, which could indicate some kind of compensatory mechanism related to presbycusis.

Age- and gender-related changes in the cochlear sympathetic system of the rat.

The sympathetic innervation projecting to the cochlea plays an important role in the auditory function, there is, however, no information about whether it is altered with advancing age. High performance liquid chromatography coupled to electrochemical detection was used to quantify both basal and noise-induced concentrations of norepinephrine (NE) in the rat cochlea. The cochlear concentration of NE was found to be independent of age in adult (3-12 months old) and aged (19 and 24 months old) males and the adult females. However, the concentrations of NE increased in aged females with respect to the younger ones, which suggests an increase in NE synthesis and a reduced NE release. Thus, a prominent gender effect emerged from this study, since the NE cochlear concentration was lower in adult females than in males, but tended to be the same level in aged animals. These modifications could be related to dramatic hormonal changes occurring in females with advancing age.