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
Direct drug delivery to the cochlea is associated with the risk of irreversible damage to the ear. In this study, liposome and polymersome nanoparticles (NPs), both formed from amphiphilic molecules (lipids in liposomes and block copolymers in polymersomes), were tested as potential tools for drug delivery to the cochlea via application onto the round window membrane in adult mice (strain C3H). One day after round window membrane application, both types of NPs labeled with fluorescent markers were identified in the spiral ganglion in all cochlear turns without producing any distinct morphological or functional damage to the inner ear. NPs were detected, although to a lesser extent, in the organ of Corti and the lateral wall. The potential of liposome and polymersome NPs as therapeutic delivery systems into the cochlea via the round window membrane was evaluated using disulfiram, a neurotoxic agent, as a model payload. Disulfiram-loaded NP delivery resulted in a significant decrease in the number of spiral ganglion cells starting 2 days postapplication, with associated pronounced hearing loss reaching 20-35 dB 2 weeks postapplication as assessed through auditory brainstem responses. No changes in hair cell morphology and function (as assessed by recording otoacoustic emissions) were detected after disulfiram-loaded NP application. No effects were observed in controls where solution of free disulfiram was similarly administered. The results demonstrate that liposome and polymersome NPs are capable of carrying a payload into the inner ear that elicits a biological effect, with consequences measurable by a functional readout.
- MeSH
- apoptóza účinky léků MeSH
- Cortiho orgán účinky léků ultrastruktura MeSH
- cytotoxiny aplikace a dávkování farmakologie MeSH
- disulfiram aplikace a dávkování farmakologie MeSH
- fenestra rotunda účinky léků metabolismus ultrastruktura MeSH
- ganglion spirale cytologie účinky léků MeSH
- kaspasa 3 metabolismus MeSH
- kochlea účinky léků metabolismus ultrastruktura MeSH
- liposomy analýza MeSH
- myši MeSH
- nanočástice analýza MeSH
- povrchově aktivní látky chemie MeSH
- systémy cílené aplikace léků metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cytotoxiny MeSH
- disulfiram MeSH
- kaspasa 3 MeSH
- liposomy MeSH
- povrchově aktivní látky MeSH
Calbindin (CB) and S100 are calcium-binding proteins expressed in the inner ear in vertebrates. Information about their developmental roles is incomplete. This study investigated the expression patterns of CB and S100 in C3H mice using immunohistochemistry, from embryonic day 11 (E11) to postnatal day 10 (P10). CB was expressed in the otocyst and vestibulocochlear ganglion (VCG) from E11. In the cochlea at E17, CB immunoreactivity clearly labeled the VCG, the outer and inner hair cells, and the stria vascularis. CB staining was also present in the vestibular sensory cells, including their nerve fibers. Two days later, to this expression pattern was added the labeling of Kölliker's organ. Early postnatal CB expression encompassed VCG neurons, auditory hair cells, their afferent nerve fibers, and cells of the cochlear lateral wall. The first signs of S100 immunostaining of cochlear and vestibular epithelial cells appeared at E14. At E17 S100 immunoreactivity was found in a restricted expression pattern in the cochlea. Immunostaining was also present in the sacculus and utriculus and their afferent fibers. The Deiters', pillar and inner hair cells, and the VCG were S100-positive from E19. Postnatally, S100 staining also appeared in the inner hair cells and Deiters' cells, in some VCG neurons, and, in addition, in the spiral limbus, the spiral prominence, and the intermediate cells of the stria vascularis. This study demonstrates that the sites of CB and S100 expression in the mouse inner ear during embryonic and early postnatal development do not overlap and signal independent developmental patterns.
- MeSH
- embryo savčí MeSH
- imunohistochemie MeSH
- kalbindiny MeSH
- kochlea embryologie růst a vývoj metabolismus MeSH
- myši inbrední C3H * MeSH
- myši MeSH
- neurony metabolismus MeSH
- novorozená zvířata MeSH
- proteiny S100 metabolismus MeSH
- S100 kalcium vázající protein G metabolismus MeSH
- stria vascularis metabolismus MeSH
- těhotenství MeSH
- tkáňová distribuce MeSH
- vláskové buňky metabolismus MeSH
- vnitřní ucho embryologie růst a vývoj metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kalbindiny MeSH
- proteiny S100 MeSH
- S100 kalcium vázající protein G MeSH
The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Sperm counts decreased to 46% (VIN-GD) and to 81% (VIN-PND), and also the acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to the control group (100%). Sperm head abnormalities increased by approximately 18% and 13%, respectively. In this connection, the expression of some genes was changed (arosome-related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, a low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups.
- MeSH
- antagonisté androgenů toxicita MeSH
- fungicidy průmyslové toxicita MeSH
- mužské pohlavní orgány účinky léků embryologie MeSH
- myši inbrední ICR MeSH
- myši MeSH
- oxazoly toxicita MeSH
- plod účinky léků MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- spermatogeneze účinky léků MeSH
- těhotenství MeSH
- tělesná hmotnost účinky léků MeSH
- testis účinky léků metabolismus MeSH
- ubikvitin metabolismus MeSH
- velikost orgánu účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antagonisté androgenů MeSH
- fungicidy průmyslové MeSH
- oxazoly MeSH
- ubikvitin MeSH
- vinclozolin MeSH Prohlížeč
The Fischer 344 rat strain has been frequently used as an animal model of rapid aging. The present study was aimed at evaluating the incidence of apoptotic cells in the inner ear of 20-24-month-old F344 rats and to correlate it with cochlear function using otoacoustic emissions. Staining with cresyl violet and the enzymatic labeling (terminal deoxynucleotidyl transferase, TdT) of fragmented DNA revealed large numbers of apoptotic cells in the marginal and basal layers of the stria vascularis and in adjacent cells of the spiral ligament. The amplitudes of distortion products otoacoustic emissions (DPOAEs), which reflect functional state of the outer hair cells, were significantly reduced or totally absent in these animals. In contrast to old F344 rats, no marked DPOAE amplitude reduction and smaller numbers of apoptotic cells were found in young 4-month-old F344 rats or in aged 24-28-month-old Long Evans rats. The accumulation of apoptotic cells, mainly in the basal layer of the stria vascularis and in adjacent cells of the spiral ligament, leads to a detachment of the stria vascularis from the spiral ligament and results in the impairment of outer hair cell function. This specific type of strial deterioration suggests that aged F344 rats can serve as an animal model of strial presbycusis.
- MeSH
- apoptóza MeSH
- buněčná smrt MeSH
- druhová specificita MeSH
- fragmentace DNA MeSH
- imunohistochemie MeSH
- kochlea cytologie růst a vývoj fyziologie MeSH
- krysa rodu Rattus MeSH
- modely u zvířat MeSH
- potkani inbrední F344 růst a vývoj fyziologie MeSH
- sluch fyziologie MeSH
- stárnutí fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hearing function in the Fischer 344 (F344) albino inbred strain of rats deteriorates with aging faster than in other strains, in spite of the small hair cell loss in old F344 animals [Popelar, J., Groh, D., Pelanova, J., Canlon, B., Syka, J., 2005. Age-related changes in cochlear and brainstem auditory function. Neurobiol. Aging, in press.]. This study was aimed at elucidating the structural changes in the inner ear of this rat strain during aging. Cochlear histopathology was examined in 20-24-month-old F344 rats and compared with that of young F344 rats (4 months) and of old rats of the Long-Evans (LE) strain. Hematoxylin/eosin staining in aged F344 rats showed degenerative changes in the organ of Corti, consisting of a damaged layer of marginal cells, reduced vascularization of the stria vascularis and a distorted tectorial membrane detached from the organ of Corti. Age-related changes in collagen distribution were observed with Masson's trichrome staining in the spiral ligament of old F344 rats. The results of immunohistochemical staining for type II collagen revealed a marked decrease in collagen fibers in the area connecting the spiral ligament and stria vascularis and a decrease in area IV fibrocytes in old F344 but not in LE rats. These findings may contribute to an explanation of the substantial hearing loss found in old F344 rats.
- MeSH
- azosloučeniny analýza MeSH
- barvicí látky analýza MeSH
- Cortiho orgán chemie MeSH
- eosin analýza MeSH
- fluorescenční barviva analýza MeSH
- hematoxylin analýza MeSH
- imunohistochemie metody MeSH
- kochlea chemie fyziologie MeSH
- kolagen typ II analýza MeSH
- kolagen analýza MeSH
- krysa rodu Rattus MeSH
- membrana tectoria chemie MeSH
- methylová zeleň analýza MeSH
- potkani inbrední F344 MeSH
- stárnutí fyziologie MeSH
- stria vascularis chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- azosloučeniny MeSH
- barvicí látky MeSH
- eosin MeSH
- fluorescenční barviva MeSH
- hematoxylin MeSH
- kolagen typ II MeSH
- kolagen MeSH
- methylová zeleň MeSH
- trichrome stain MeSH Prohlížeč
The Splotch mouse, a Pax 3 mutation, represents a model of Waardenburg syndrome I. We show that the homozygous Splotch mutation (Sp(2H)) is associated with severe defects that prevent the formation of the cochlea and vestibulo-cochlear ganglion. To clarify the role of Pax 3 in inner ear formation, we examined the expression of polysialic acid (PSA) associated with neural cell adhesion molecule (NCAM). In accordance with the occurrence of phenotypic abnormalities, PSA NCAM was expressed early in otocyst development in the otic epithelium and the vestibulo-cochlear anlage. During the period of vestibular and cochlear ganglia formation, PSA NCAM expression was decreased. In the late phase of embryonic development, the expression of calcium binding proteins (S100) in the vestibulo-cochlear ganglion was also decreased. Minor differences in S100 immunostaining were found postnatally between the cochleas of heterozygous and wild type animals.
- MeSH
- buněčná smrt genetika MeSH
- DNA vazebné proteiny genetika metabolismus MeSH
- embryo savčí MeSH
- imunohistochemie metody MeSH
- koncové značení zlomů DNA in situ MeSH
- kyseliny sialové metabolismus MeSH
- molekula buněčné adheze nervové L1 metabolismus MeSH
- mutace * MeSH
- mutantní kmeny myší anatomie a histologie růst a vývoj metabolismus MeSH
- myši MeSH
- novorozená zvířata MeSH
- počet buněk metody MeSH
- proteiny S100 metabolismus MeSH
- transkripční faktor PAX3 MeSH
- transkripční faktory paired box MeSH
- transkripční faktory genetika metabolismus MeSH
- věkové faktory MeSH
- vnitřní ucho anatomie a histologie růst a vývoj metabolismus MeSH
- vývojová regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- DNA vazebné proteiny MeSH
- kyseliny sialové MeSH
- molekula buněčné adheze nervové L1 MeSH
- Pax3 protein, mouse MeSH Prohlížeč
- polysialyl neural cell adhesion molecule MeSH Prohlížeč
- proteiny S100 MeSH
- transkripční faktor PAX3 MeSH
- transkripční faktory paired box MeSH
- transkripční faktory MeSH
Hybrid sterility is a common postzygotic reproductive isolation mechanism that appears in the early stages of speciation of various organisms. Mus musculus musculus and Mus musculus domesticus represent two recently separated mouse subspecies particularly suitable for genetic studies of hybrid sterility. Here we show that the introgression of Chr X of M. m. musculus origin (PWD/Ph inbred strain, henceforth PWD) into the genetic background of the C57BL/6J (henceforth B6) inbred strain (predominantly of M. m. domesticus origin) causes male sterility. The X-linked hybrid sterility is associated with reduced testes weight, lower sperm count, and morphological abnormalities of sperm heads. The analysis of recombinant Chr Xs in sterile and fertile males as well as quantitative trait locus (QTL) analysis of several fertility parameters revealed an oligogenic nature of the X-linked hybrid sterility. The Hstx1 locus responsible for male sterility was mapped near DXMit119 in the central part of Chr X. To ensure full sterility, the PWD allele of Hstx1 has to be supported with the PWD allelic form of loci in at least one proximal and/or one distal region of Chr X. Mapping and cloning of Hstx1 and other genes responsible for sterility of B6-X PWD Y B6 males could help to elucidate the special role of Chr X in hybrid sterility and consequently in speciation.
- MeSH
- akrozomální reakce MeSH
- alely MeSH
- chromozom X * MeSH
- fenotyp MeSH
- genetická vazba * MeSH
- genetické markery MeSH
- genotyp MeSH
- křížení genetické MeSH
- lokus kvantitativního znaku MeSH
- monoklonální protilátky chemie MeSH
- mužská infertilita genetika MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- rekombinantní proteiny chemie MeSH
- sexuální faktory MeSH
- spermie patologie MeSH
- testis patologie MeSH
- ženská infertilita genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- genetické markery MeSH
- monoklonální protilátky MeSH
- rekombinantní proteiny MeSH
The effect of genistein (GEN) and diethylstilbestrol (DES) on body weight, weight of different organs, sperm acrosomal status and in vivo fertility of CD1 mice was tested in a multigenerational study. The adult parental generation of mice and F1 and F2 generations were exposed to selected drugs for all their life. GEN had effect on different body parameters of 30-day-old mice, but not of adult mice in the first generation. Contrary to that, treatment by DES had a strong effect on body weight, other body parameters and on the levels of serum hormones. In the first generation only sterile pairs of mice were observed. Monoclonal antibody against mouse intra-acrosomal sperm protein was used for analysis of the acrosome state and as biomarkers of sperm damage. In the control groups, about 93% of acrosome-reacted sperm was found, acrosome staining decreased to 78-84% (P<0.01). However, the GEN had no effect on fertility of CD1 mice. On the other hand, the fertility of mice exposed to DES was disrupted, especially in the first generation.
- MeSH
- akrozom fyziologie MeSH
- akrozomální reakce MeSH
- diethylstilbestrol aplikace a dávkování MeSH
- folikuly stimulující hormon krev MeSH
- genistein aplikace a dávkování MeSH
- myši MeSH
- ovarium anatomie a histologie MeSH
- spermie fyziologie ultrastruktura MeSH
- tělesná hmotnost * MeSH
- testis anatomie a histologie MeSH
- testosteron krev MeSH
- velikost orgánu MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- diethylstilbestrol MeSH
- folikuly stimulující hormon MeSH
- genistein MeSH
- testosteron MeSH
BACKGROUND: The splotch (Sp(2H)), Pax-3 mutant mice are characterized by neurulation defects, neural crest deficiencies, and altered somitogenesis. A link connecting all morphological abnormalities in the Pax-3 homozygous embryos is the composition of extracellular matrix and misexpression of cell adhesion molecules. The neural cell adhesion molecule (NCAM) is one of the Pax-3 target genes. Its adhesive properties depend on the attached polysialic acid (PSA). We have studied whether NCAM sialylation has been affected in the Pax-3 mutant embryos. METHODS: Genotyping of embryos was determined using polymerase chain reaction. The periodate-resorcinol method was used for quantitative determination of sialic acid. Immunoblotting was used to detect sialylated NCAM isoforms and sialic acids on the blot. This antibody was also used to detect PSA-NCAM spatial expression. RESULTS: Quantitative determination of sialic acid at days 10.5-13.5 showed decreased sialic acid content in Sp(2H) homozygotes. The results of both techniques used in evaluating the expression of PSA-NCAM isoforms in our study indicate that the 180-kDa isoform is decreased in homozygous Splotch (Sp(2H)) embryos. Immunohistochemistry showed decreased staining in the neural tube, ganglion VIII, (day 13.5), frontal lobe, olfactory bulb, and neuroblastic retinal cells (day 18.5). CONCLUSIONS: PSA-NCAM is present in Sp(2H) embryos of all genotypes, starting on developmental day 9.5. Sialylation of the 180-kDa NCAM isoform begins to decrease on day 12.5 in Sp(2H) homozygotes. Reduced NCAM sialylation could contribute to the decreased migration of cells in the sensory organs.
- MeSH
- DNA primery MeSH
- DNA vazebné proteiny genetika MeSH
- embryo savčí metabolismus MeSH
- homozygot * MeSH
- kyselina N-acetylneuraminová metabolismus MeSH
- molekuly buněčné adheze nervové metabolismus MeSH
- myši MeSH
- sekvence nukleotidů MeSH
- transkripční faktor PAX3 MeSH
- transkripční faktory paired box MeSH
- transkripční faktory genetika MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA primery MeSH
- DNA vazebné proteiny MeSH
- kyselina N-acetylneuraminová MeSH
- molekuly buněčné adheze nervové MeSH
- Pax3 protein, mouse MeSH Prohlížeč
- transkripční faktor PAX3 MeSH
- transkripční faktory paired box MeSH
- transkripční faktory MeSH