The development, maturation, and maintenance of the inner ear are governed by temporal and spatial expression cascades of transcription factors that form a gene regulatory network. ISLET1 (ISL1) may be one of the major players in this cascade, and in order to study its role in the regulation of inner ear development, we produced a transgenic mouse overexpressing Isl1 under the Pax2 promoter. Pax2-regulated ISL1 overexpression increases the embryonic ISL1(+) domain and induces accelerated nerve fiber extension and branching in E12.5 embryos. Despite these gains in early development, the overexpression of ISL1 impairs the maintenance and function of hair cells of the organ of Corti. Mutant mice exhibit hyperactivity, circling behavior, and progressive age-related decline in hearing functions, which is reflected in reduced otoacoustic emissions (DPOAEs) followed by elevated hearing thresholds. The reduction of the amplitude of DPOAEs in transgenic mice was first detected at 1 month of age. By 6-9 months of age, DPOAEs completely disappeared, suggesting a functional inefficiency of outer hair cells (OHCs). The timing of DPOAE reduction coincides with the onset of the deterioration of cochlear efferent terminals. In contrast to these effects on efferents, we only found a moderate loss of OHCs and spiral ganglion neurons. For the first time, our results show that the genetic alteration of the medial olivocochlear (MOC) efferent system induces an early onset of age-related hearing loss. Thus, the neurodegeneration of the MOC system could be a contributing factor to the pathology of age-related hearing loss.
- Klíčová slova
- Age-related hearing loss, Islet1 transcription factor, Medial olivocochlear efferent system, Outer hair cells, Transgenic mouse,
- MeSH
- analýza přežití MeSH
- embryo savčí metabolismus patologie MeSH
- ganglion spirale patologie MeSH
- kochlea inervace patologie patofyziologie MeSH
- messenger RNA genetika metabolismus MeSH
- molekulární motory metabolismus MeSH
- myši transgenní MeSH
- nedoslýchavost patologie patofyziologie MeSH
- neurony eferentní MeSH
- otoakustické emise spontánní MeSH
- počet buněk MeSH
- proteiny s homeodoménou LIM metabolismus MeSH
- sluchový práh MeSH
- stárnutí patologie MeSH
- transkripční faktor PAX2 metabolismus MeSH
- transkripční faktory metabolismus MeSH
- vnější vláskové buňky patologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- insulin gene enhancer binding protein Isl-1 MeSH Prohlížeč
- messenger RNA MeSH
- molekulární motory MeSH
- Pax2 protein, mouse MeSH Prohlížeč
- Pres protein, mouse MeSH Prohlížeč
- proteiny s homeodoménou LIM MeSH
- transkripční faktor PAX2 MeSH
- transkripční faktory MeSH
Ischemia can contribute to the inner ear pathology and hearing loss. To determine the susceptibility of inner and outer hair cells (IHCs/OHCs) to ischemic and post-ischemic period, we used organotypic cultures of the organ of Corti isolated from P3 rats as an in vitro model of inner ear ischemia (oxygen-glucose deprivation, OGD). We identified the hair cells (HCs) by phalloidin staining. The cells with damaged cellular membrane integrity were identified by propidium iodide (PI)-exclusion assay. The cells with fragmented chromosomal DNA were detected by TUNEL assay. Organotypic cultures were subjected to a mild (3 h duration) or severe (4 h duration) OGD, followed by a recovery period of 21 h and 20 h, respectively. Mild OGD induced a loss of 10-20% HCs, whereas severe OGD induced loss of 35% HCs. We confirmed that OHCs are less vulnerable to OGD than IHCs. Of all missing OHCs, 80-90% was lost during the OGD period and 10-20% during the recovery period. In contrast, the loss of IHCs was equal during both experimental periods. The OGD period was mainly associated with PI-positive nuclei. TUNEL-positive nuclei were a minor fraction during the OGD period and increased during the recovery period, indicating the progression of DNA fragmentation. Our results implicate a differential susceptibility of IHCs and OHCs during and after ischemia-like insult, which may be of therapeutic consequence.
- MeSH
- apoptóza MeSH
- barvení a značení metody MeSH
- časové faktory MeSH
- fragmentace DNA MeSH
- glukosa metabolismus MeSH
- hyperglykemie metabolismus patologie MeSH
- hypoxie buňky MeSH
- koncové značení zlomů DNA in situ MeSH
- krysa rodu Rattus MeSH
- kyslík metabolismus MeSH
- novorozená zvířata MeSH
- orgánové kultury - kultivační techniky MeSH
- permeabilita buněčné membrány MeSH
- potkani Wistar MeSH
- vnější vláskové buňky metabolismus patologie MeSH
- vnitřní vláskové buňky metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- glukosa MeSH
- kyslík MeSH