Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects.

Transplantation of hematopoietic stem cells (HSCs) is regarded to be a potential approach for promoting repair of damaged organs. Here, we investigated the influence of hematopoietic stem cells on progressive hair cell degeneration after transient cochlear ischemia in gerbils. Transient cochlear ischemia was produced by extracranial occlusion of the bilateral vertebral arteries just before their entry into the transverse foramen of the cervical vertebra. Intrascalar injection of HSCs prevented ischemia-induced hair cell degeneration and ameliorated hearing impairment. We also showed that the protein level of glial cell line-derived neurotrophic factor (GDNF) in the organ of Corti was upregulated after cochlear ischemia and that treatment with HSCs augmented this ischemia-induced upregulation of GDNF. A tracking study revealed that HSCs injected into the cochlea were retained in the perilymphatic space of the cochlea, although they neither transdifferentiated into cochlear cell types nor fused with the injured hair cells after ischemia, suggesting that HSCs had therapeutic potential possibly through paracrine effects. Thus, we propose HSCs as a potential new therapeutic strategy for hearing loss.

Adenovirus-mediated overexpression of a gene prevents hearing loss and progressive inner hair cell loss after transient cochlear ischemia in gerbils.

The use of adenoviral vectors has recently provided a novel strategy for direct gene transfer into the cochlea. In this study, we assessed the utility of an adenoviral vector expressing glial-cell-derived neurotrophic factor (GDNF) in ischemia-reperfusion injury of the gerbil cochlea. The vector was injected through the round window 4 days before ischemic insult. The distribution of a reporter transgene was confirmed throughout the cochlea from the basal to the apical turn and Western blot analysis indicated significant upregulation of GDNF protein 11 days following virus inoculation. Hearing ability was assessed by sequentially recording compound action potentials (CAP), and the degree of hair cell loss in the organ of Corti was evaluated in specimens stained with rhodamine-phalloidin and Hoechst 33342. On the seventh day of ischemia, the CAP threshold shift and inner hair cell loss were remarkably suppressed in the Ad-GDNF group compared with the control group. These results suggest that adenovirus-mediated overexpression of GDNF is useful for protection against hair cell damage, which otherwise eventually occurs after transient ischemia of the cochlea.

Hypothermia reduces glutamate efflux in perilymph following transient cochlear ischemia.

The effect of hypothermia on ischemic injury of the cochlea in gerbils was studied with particular regard to glutamate efflux in the perilymph. Under normothermic conditions interruption of the blood supply to the cochlea for 15 min caused a remarkable elevation of the compound action potential (CAP) threshold, and an increase in perilymphatic glutamate. The CAP threshold recovered to some extent with reperfusion, but not to preischemic levels. CAP thresholds, under hypothermic conditions and with reperfusion, recovered promptly to near pre-ischemic levels, while glutamate concentration did not change. These results, together with electron microscopy studies, suggest that hypothermia prevents hearing loss primarily through reduction of glutamate efflux at the synopses between inner hair cells and primary afferent auditory neurons.

Hypothermia prevents hearing loss and progressive hair cell loss after transient cochlear ischemia in gerbils.

The effects of hypothermia on ischemia-reperfusion injury of the cochlea were studied in gerbils. Hearing was assessed by sequentially recording compound action potentials before, during and after the ischemia. The degree of hair cell loss in the organ of Corti was evaluated in specimens stained with rhodamine-phalloidin and the dye Hoechst 33342. Ischemic insult was applied to the animals by occluding the bilateral vertebral arteries for 15 min under normothermic or hypothermic (rectal temperature 32 degrees C) conditions. Interruption of the blood supply to the cochlea caused a tremendous increase in the compound action potential threshold, which usually recovered to some extent with reperfusion. In the ischemia/normothermic group, the threshold did not return to the pre-ischemic level. The average increase in the threshold seven days after ischemia was 20.0 dB. Histologically, the hair cell loss increased gradually until four days after the ischemic insult. On the seventh day, the mean loss of inner and outer hair cells at the basal turn was 31.1 % and 2.4 %, respectively. In the ischemia/hypothermic group, the threshold returned to the pre-ischemic level within 30 min after reperfusion and remained stable thereafter. The mean loss of inner and outer hair cells on the seventh day was 0.1 % and 0.2 %, respectively. These results indicate that hypothermia can prevent inner ear damage, which otherwise occurs after transient ischemia of the cochlea.

Hearing loss and glutamate efflux in the perilymph following transient hindbrain ischemia in gerbils.

The mechanism underlying ischemia-induced hearing loss was studied in gerbils with transient hindbrain ischemia. Occlusion of the vertebral arteries caused an increase in the concentration of glutamate in the perilymph and elevated the compound action potential (CAP) threshold to 24.6 dB at 5 minutes. the CAP threshold subsequently recovered on reperfusion, gradually reaching 8.3 dB 120 minutes after reperfusion. Under electron microscopy, afferent dendrites of the cochlear nerve in contact with inner hair cells exhibited abnormal swelling 5 minutes after ischemia/reperfusion. These morphological changes were not observed in cochleas treated with an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate-type glutamate receptor antagonist, 6-7-dinitroquinoxaline-2,3-dione (DNQX), before hindbrain ischemia; an N-methyl-D-aspartate (NMDA)-type receptor antagonist, D-2-amino-5-phosphonopentanoate (D-AP5), was ineffective. Moreover, the histopathological alterations noted 5 minutes after reperfusion were spontaneously ameliorated 120 minutes after ischemia/reperfusion. These findings suggest that the ischemia-induced increase in extracellular glutamate concentration with subsequent activation of AMPA/kainate receptors is responsible for neurite degeneration and hearing loss in the early stages following transient hindbrain ischemia.

Exacerbation of noise-induced hearing loss in mice lacking the glutamate transporter GLAST.

Acoustic overstimulation is one of the major causes of hearing loss. Glutamate is the most likely candidate neurotransmitter for afferent synapses in the peripheral auditory system, so it was proposed that glutamate excitotoxicity may be involved in noise trauma. However, there has been no direct evidence that noise trauma is caused by excessive release of glutamate from the inner hair cells (IHCs) during sound exposure because studies have been hampered by powerful glutamate uptake systems in the cochlea. GLAST is a glutamate transporter highly expressed in the cochlea. Here we show that after acoustic overstimulation, GLAST-deficient mice show increased accumulation of glutamate in perilymphs, resulting in exacerbation of hearing loss. These results suggest that GLAST plays an important role in keeping the concentration of glutamate in the perilymph at a nontoxic level during acoustic overstimulation. These findings also provide further support for the hypothesis that IHCs use glutamate as a neurotransmitter.

Efflux of glutamate into the perilymph of the cochlea following transient ischemia in the gerbil.

Using a microdialysis technique followed by an enzyme cycling analysis, we measured changes in the glutamate levels in the perilymph of gerbil cochleae before, during and after transient ischemic insult. The basal glutamate level in perilymph was 0.35 +/- 0.22 pmol/microl. An almost immediate and continuous rise in the level of glutamate occurred after the ischemic insult, which advanced even further after recirculation; the average concentration was higher than 40 pmol/microl 55 min after recirculation. The compound action potentials (CAP) monitoring the auditory function totally disappeared after ischemic insult. However, CAP reappeared after recirculation; the threshold for acoustic stimulation was higher than that observed at the pre-ischemic state.

[Computerized database system for pure tone audiometry].

The management and storage of pure tone audiometry data creates a major problem for otolaryngological clinics. We have devised a new computerized database system to overcome this problem. The hardware involved includes multiple audiometers and computers interfaced with a glassfiber local area network (Pi-Net). The software used is database 3 plus, a popular database language and utility software package. Programs were written for data entry, data transfer, reference, entry of patient ID, and evaluation of tympanoplasty. The following benefits of the system are noted. 1) Data entry is possible at every terminal (audiometer and computer). 2) Storage capacity is sufficient for more than 10 years of operation. 3) All data can be retrieved rapidly at any terminal. Evaluations of pre- and post-tympanoplasty hearing loss are facilitated by this system. This system is very effective for follow up of hearing disorders.