RESUMO
Control over ionic composition and volume of the inner ear luminal fluid endolymph is essential for normal hearing and balance. Mice deficient in either the EphB2 receptor tyrosine kinase or the cognate transmembrane ligand ephrin-B2 (Efnb2) exhibit background strain-specific vestibular-behavioral dysfunction and signs of abnormal endolymph homeostasis. Using various loss-of-function mouse models, we found that Efnb2 is required for growth and morphogenesis of the embryonic endolymphatic epithelium, a precursor of the endolymphatic sac (ES) and duct (ED), which mediate endolymph homeostasis. Conditional inactivation of Efnb2 in early-stage embryonic ear tissues disrupted cell proliferation, cell survival, and epithelial folding at the origin of the endolymphatic epithelium. This correlated with apparent absence of an ED, mis-localization of ES ion transport cells relative to inner ear sensory organs, dysplasia of the endolymph fluid space, and abnormally formed otoconia (extracellular calcite-protein composites) at later stages of embryonic development. A comparison of Efnb2 and Notch signaling-deficient mutant phenotypes indicated that these two signaling systems have distinct and non-overlapping roles in ES/ED development. Homozygous deletion of the Efnb2 C-terminus caused abnormalities similar to those found in the conditional Efnb2 null homozygote. Analyses of fetal Efnb2 C-terminus deletion heterozygotes found mis-localized ES ion transport cells only in the genetic background exhibiting vestibular dysfunction. We propose that developmental dysplasias described here are a gene dose-sensitive cause of the vestibular dysfunction observed in EphB-Efnb2 signaling-deficient mice.
Assuntos
Orelha Interna/metabolismo , Saco Endolinfático/metabolismo , Efrina-B2/genética , Epitélio/metabolismo , Animais , Proliferação de Células , Sobrevivência Celular/genética , Orelha Interna/embriologia , Embrião de Mamíferos/citologia , Embrião de Mamíferos/embriologia , Embrião de Mamíferos/metabolismo , Saco Endolinfático/embriologia , Saco Endolinfático/ultraestrutura , Efrina-B2/metabolismo , Epitélio/embriologia , Epitélio/ultraestrutura , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Confocal , Microscopia Eletrônica de Varredura , Morfogênese/genética , Gravidez , Receptores Notch/genética , Receptores Notch/metabolismo , Transdução de Sinais/genética , Fatores de TempoRESUMO
Injury to the endolymphatic sac may play an important role in the pathogenesis of Ménière's disease, an inner ear disorder characterized by hearing loss, tinnitus and attacks of vertigo. Isoimmunization of 16 inbred Lewis rats with a crude endolymphatic sac extract and complete Freund's adjuvant induced hyperactivity of the endolymphatic sac. One group of rats was immunized by a single dose whereas a second group was immunized twice. Control animals were injected with Freund's adjuvant in saline only. Serum was collected from all rats by the end of the study and harvested autoantibodies were tested by immunohistochemistry. The endolymphatic sacs were investigated by transmission electron microscopy. Endolymphatic sac stimulation was observed in all immunized rats. Based on detailed ultrastructural observations, the degree of reactivity seemed proportional to the number of injections and the extent of immunization. Moreover, the ribosome-rich cells seemed hyperactive with an extravagant content of intracellular components: numerous rough endoplasmic reticulum and free ribosomes, morphological signs of extensive endo- and exocytosis, vesicles of material with a density similar to the homogeneous substance of which many were observed to fuse with primary lysozymes. Basolateral foldings were numerous and in the subepithelial capillaries formation of multiple and apposing fenestrations were observed. No endolymphatic sac stimulation was observed in the control animals. Specific ribosome-rich cell alterations identical to those present in the endolymphatic sac of Ménière's disease were observed 21 days after the first immunization. The observations suggest that either an autoantigen or a trophic factor, capable of inducing a hyperactivity of the ribosome-rich cells and an imbalance of the homogeneous substance metabolism, exists in the endolymphatic sac of the rat.
Assuntos
Saco Endolinfático/fisiopatologia , Doença de Meniere/fisiopatologia , Extratos de Tecidos/farmacologia , Adjuvantes Imunológicos/farmacologia , Animais , Autoantígenos/imunologia , Modelos Animais de Doenças , Saco Endolinfático/patologia , Saco Endolinfático/ultraestrutura , Retículo Endoplasmático Rugoso/patologia , Retículo Endoplasmático Rugoso/ultraestrutura , Adjuvante de Freund/farmacologia , Imunização/métodos , Masculino , Doença de Meniere/imunologia , Doença de Meniere/patologia , Microscopia Eletrônica de Transmissão , Mitocôndrias/patologia , Mitocôndrias/ultraestrutura , Ratos , Ratos Endogâmicos Lew , Ratos Wistar , Ribossomos/patologia , Ribossomos/ultraestrutura , Especificidade da Espécie , Junções Íntimas/patologia , Junções Íntimas/ultraestrutura , Extratos de Tecidos/imunologiaRESUMO
The middle ear infection is the most common childhood infection. In order to elucidate the cell and molecular mechanisms involved in bacterial recognition and innate immune response, we have established a stable human middle ear cell line, which has contributed to the current knowledge concerning the molecular pathogenesis of the middle ear infection. The inner ear, a sensory organ responsible for hearing and balance, is filled with inner ear fluid, and disturbance of the fluid homeostasis results in dizziness and hearing impairment. It has been suggested that the endolymphatic sac (ES) may play a critical role in the fluid homeostasis of the inner ear. We have established a stable human ES cell line and are undertaking cell and molecular characterization of this cell line.
Assuntos
Orelha Média/citologia , Saco Endolinfático/citologia , Linhagem Celular , Orelha Média/ultraestrutura , Saco Endolinfático/ultraestrutura , Células Epiteliais/fisiologia , HumanosRESUMO
The inner ear is a fluid-filled closed-epithelial structure whose function requires maintenance of an internal hydrostatic pressure and fluid composition. The endolymphatic sac (ES) is a dead-end epithelial tube connected to the inner ear whose function is unclear. ES defects can cause distended ear tissue, a pathology often seen in hearing and balance disorders. Using live imaging of zebrafish larvae, we reveal that the ES undergoes cycles of slow pressure-driven inflation followed by rapid deflation. Absence of these cycles in lmx1bb mutants leads to distended ear tissue. Using serial-section electron microscopy and adaptive optics lattice light-sheet microscopy, we find a pressure relief valve in the ES comprised of partially separated apical junctions and dynamic overlapping basal lamellae that separate under pressure to release fluid. We propose that this lmx1-dependent pressure relief valve is required to maintain fluid homeostasis in the inner ear and other fluid-filled cavities.
Assuntos
Saco Endolinfático/ultraestrutura , Audição/fisiologia , Larva/ultraestrutura , Fatores de Transcrição/genética , Proteínas de Peixe-Zebra/genética , Animais , Animais Geneticamente Modificados , Embrião não Mamífero , Saco Endolinfático/anatomia & histologia , Saco Endolinfático/fisiologia , Feminino , Expressão Gênica , Homeostase/fisiologia , Pressão Hidrostática , Hibridização in Situ Fluorescente , Larva/anatomia & histologia , Larva/fisiologia , Masculino , Microscopia Eletrônica , Mutação , Imagem com Lapso de Tempo , Fatores de Transcrição/metabolismo , Peixe-Zebra , Proteínas de Peixe-Zebra/metabolismoRESUMO
Background: The endolymphatic sac (ES) is endowed with a multitude of white blood cells that may trap and process antigens that reach the inner ear from nearby infection-prone areas, it thus serves as an immunologic defense organ. The human ES, and unexpectedly the rest of the inner ear, has been recently shown to contain numerous resident macrophages. In this paper, we describe ES macrophages using super-resolution structured fluorescence microscopy (SR-SIM) and speculate on these macrophages' roles in human inner ear defense. Material and Methods: After ethical permission was obtained, human vestibular aqueducts were collected during trans-labyrinthine surgery for acoustic neuroma removal. Tissues were placed in fixative before being decalcified, rapidly frozen, and cryostat sectioned. Antibodies against IBA1, cytokine fractalkine (CX3CL1), toll-like receptor 4 (TLR4), cluster of differentiation (CD)68, CD11b, CD4, CD8, and the major histocompatibility complex type II (MHCII) were used for immunohistochemistry. Results: A large number of IBA1-positive cells with different morphologies were found to reside in the ES; the cells populated surrounding connective tissue and the epithelium. Macrophages interacted with other cells, showed migrant behavior, and expressed immune cell markers, all of which suggest their active role in the innate and adaptive inner ear defense and tolerance. Discussion: High-resolution immunohistochemistry shows that antigens reaching the ear may be trapped and processed by an immune cell machinery located in the ES. Thereby inflammatory activity may be evaded near the vulnerable inner ear sensory structures. We speculate on the immune defensive link between the ES and the rest of the inner ear.
Assuntos
Orelha Interna/imunologia , Saco Endolinfático/imunologia , Biomarcadores , Proteínas de Ligação ao Cálcio , Comunicação Celular/imunologia , Quimiocina CX3CL1/genética , Cóclea/imunologia , Cóclea/metabolismo , Cóclea/ultraestrutura , Proteínas de Ligação a DNA/genética , Orelha Interna/diagnóstico por imagem , Orelha Interna/ultraestrutura , Saco Endolinfático/diagnóstico por imagem , Saco Endolinfático/ultraestrutura , Expressão Gênica , Humanos , Imunidade , Imuno-Histoquímica , Imunofenotipagem , Macrófagos/imunologia , Macrófagos/metabolismo , Proteínas dos Microfilamentos , Conformação Molecular , Microtomografia por Raio-XRESUMO
In an investigation aimed at clarifying the mechanism of crystal dissolution of the calcium carbonate lattice in otoconia (the mineral particles embedded in the otolithic membrane) of the endolymphatic sac (ELS) of the bullfrog, cDNAs encoding the A- and E-subunits of bullfrog vacuolar proton-pumping ATPase (V-ATPase) were cloned and sequenced. The cDNA of the A-subunit consisted of an 11-bp 5'-untranslated region (UTR), a 1,854-bp open reading frame (ORF) encoding a protein comprising 617 amino acids with a calculated molecular mass of 68,168 Da, and a 248-bp 3'-UTR followed by a poly(A) tail. The cDNA of the E-subunit consisted of a 72-bp 5'-UTR, a 681-bp ORF encoding a protein of 226 amino acids with a calculated molecular mass of 26,020 Da, and a 799-bp 3'-UTR followed by a poly(A) tail. Western blot and immunofluorescence analyses using specific anti-peptide antisera against the V-ATPase A- and E-subunits revealed that these subunits were present in the ELS, urinary bladder, skin, testes, and kidneys. In the ELS, positive cells were scattered in the follicular epithelium which, as revealed by electron microscopy, corresponds to the location of mitochondria-rich cells. These findings suggest that V-ATPase, including the A- and E-subunits, exists in mitochondria-rich cells of the ELS, which might be involved in dissolution of the calcium carbonate crystals in the lumen of the ELS.
Assuntos
DNA Complementar/química , Saco Endolinfático/enzimologia , Rana catesbeiana , ATPases Vacuolares Próton-Translocadoras/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Clonagem Molecular , Saco Endolinfático/ultraestrutura , Epitélio/enzimologia , Imunofluorescência , Regulação Enzimológica da Expressão Gênica , Dados de Sequência Molecular , Rana catesbeiana/genética , Rana catesbeiana/metabolismo , ATPases Vacuolares Próton-Translocadoras/químicaRESUMO
INTRODUCTION: Recent studies suggest that the human endolymphatic sac (ES) may have multiple functions, including an ion-transport capacity comparable to the kidney, an immunological capacity and a possible natriuretic capacity. Further, there have been speculations of a yet undefined role in intracranial pressure homeostasis. The anatomical location towards the sigmoid sinus would suggest a possible endo- and/or paracrine signaling. However, neuronal connections may also apply, but it remains very scarcely explored in the human ES. STUDY DESIGN: DNA micro-arrays and immunohistochemistry were used for analyses of fresh human ES tissue samples. METHODS: A total of 30 tissue samples from the human ES were obtained during translabyrinthine surgery for vestibular schwannoma. Microarray technology was used to investigate tissue sample gene expression, using adjacent dura mater as control. The expression of genes specific for neuronal signaling was determined and results for selected key molecules verified by immunohistochemistry. Transmission electron microscopy was used for ultrastructural analysis. RESULTS: For the transmission electron microscopy analysis, a direct innervation of the ES was observed with unmyelinated fibers imbedded in the ES epithelial lining. The microarrays confirmed, that several molecules involved in neuronal signaling were found expressed significantly in the ES DNA profile, such as the Cholecystokinin peptide and related receptors, Dopamine receptors 2 and 5, vesicular monoamine transporter 2 (VMAT2), plasma monoamine transporter (PMAT), and Serotonin 1D. All peptides were verified by immunohistochemistry. CONCLUSIONS: Based on global gene expression profiling and immuno-histochemical labeling, we conclude that the human ES expresses neuropeptide receptors and monoamine transporters. Combined with the ultrastructural demonstration of unmyelinated axons imbedded within the epithelial lining, the findings suggest that neuro-signaling mechanisms are involved in functions exerted by the ES.
Assuntos
Saco Endolinfático/inervação , Saco Endolinfático/metabolismo , Saco Endolinfático/ultraestrutura , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Fibras Nervosas/ultraestrutura , Receptores de Neurotransmissores/biossíntese , TranscriptomaRESUMO
The endolymphatic sac (ES) is believed to play an important role in maintaining homeostasis in the inner ear by the absorption and endocytosis of endolymph. Megalin is a 600-kDa multiligand endocytic receptor expressed in certain types of absorptive epithelia including kidney proximal tubules. We analyzed the immunoreactivity for megalin in rat ES by immunofluorescence, immunogold electron microscopy, and immunoblotting. With immunostaining, the luminal substances of the ES were strongly stained for megalin. Megalin was also localized in luminal macrophage-like cells and both types of epithelial cell (mitochondria-rich cells and ribosome-rich cells). In these cells, the megalin was localized in the lumen of endosomes, but was not membrane associated. This localization pattern indicates that the megalin in these cells is not a membrane receptor, but merely one of the constituents that are endocytosed from the lumen of the ES. Immunoblotting indicated that the megalin in the ES is a 210-kDa molecule lacking a cytoplasmic domain. This suggests that the megalin in the ES may be a soluble form, different from the 600-kDa membrane-bound receptor expressed in kidneys. Taken together, it is likely that the megalin in the ES lumen is a soluble component and may be endocytosed by the ES epithelial cells. Furthermore, we found that the tectorial membrane, an acellular structure in the cochlea, gave a strong megalin immunoreaction. Since the cochlea is connected to the ES, the megalin may be transported alone or with the components of the tectorial membrane from the cochlea to the ES lumen through longitudinal flow.
Assuntos
Orelha Interna/metabolismo , Endolinfa/fisiologia , Saco Endolinfático/metabolismo , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Animais , Especificidade de Anticorpos , Orelha Interna/citologia , Eletroforese em Gel de Poliacrilamida , Endocitose/fisiologia , Saco Endolinfático/ultraestrutura , Células Epiteliais/metabolismo , Células Epiteliais/ultraestrutura , Immunoblotting , Imuno-Histoquímica , Rim/química , Macrófagos/metabolismo , Macrófagos/ultraestrutura , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Imunoeletrônica , Microssomos/química , Especificidade de Órgãos , Ratos , Ratos Sprague-Dawley , Membrana Tectorial/metabolismo , Membrana Tectorial/ultraestruturaRESUMO
CONCLUSION: Twenty-five rats were challenged by an immunologic attack of the endolymphatic sac. After 6 months, distortion product oto-acoustic emissions (DPOAE) revealed a dysfunction of the outer hair cells and immunological active cells were observed in the endolymphatic sac. This information could contribute to the understanding of Ménière's disease. OBJECTIVES: This study investigated if an autoimmune challenge of the endolymphatic sac could affect DPOAE output measurements in rats. Also, a potential autoimmune cell infiltration of the endolymphatic sac was investigated. METHODS: Eighteen Lewis rats were immunized with a crude endolymphatic sac extract in complete Freund's adjuvant. Seven control animals were injected with Freund's adjuvant in saline. Cochlear damage was estimated by DPOAE dynamics 3 weeks and 6 months after the immunization. Infiltrative cells in the endolymphatic sac were investigated with transmission electron microscopy. RESULTS: The hearing assessment 6 months after immunization revealed a reduction of the DPOAE, on the full range of frequencies (2-63 kHz) in an average of the mean, of 2 dB ± 1.1 in the immunized group compared to the controls (p < 0.05). The same test showed a 2.5 dB decrease from 2 to 5 kHz (p < 0.01). Immunological active cells were observed in the endolymphatic sac in most of the immunized rats.
Assuntos
Autoimunidade , Saco Endolinfático/ultraestrutura , Doença de Meniere/imunologia , Emissões Otoacústicas Espontâneas/fisiologia , Estimulação Acústica , Animais , Cóclea/ultraestrutura , Modelos Animais de Doenças , Saco Endolinfático/fisiopatologia , Doença de Meniere/patologia , Doença de Meniere/fisiopatologia , Microscopia Eletrônica de Transmissão , Ratos , Ratos Endogâmicos LewRESUMO
Previously, we obtained a protein that has considerable amino acid sequence homology with secretory phospholipase A(2) (PLA(2)) from a bullfrog pituitary fraction obtained during the purification of thyrotropin (TSH). Subsequently, partial amino acid sequence (N-terminal 45 amino acid residues) analysis revealed this protein to be identical to the N-terminal amino acid sequence of otoconin-22, the major protein of aragonitic otoconia in the Xenopus saccule. In this study we developed an antibody against the N-terminal peptide of the bullfrog protein and applied it for immunocytochemical study of the pituitary and its surrounding tissue. Western blotting analysis showed that this antibody recognizes a 20.4-kD protein that has a molecular mass close to that of otoconin-22. Immunohistochemical reaction with the antibody was not found in any anterior pituitary cells but was intense in the monolayer epithelial cells of the endolymphatic sac surrounding the pituitary gland, which is a major storage site of calcium carbonate in amphibians. An electron microscopic study revealed that the cuboidal cells in the endolymphatic sac contained large, polymorphic secretory granules in their apical cytoplasm. Immunogold particles indicating the presence of a PLA(2)-like protein were observed predominately in these secretory granules. These findings support the view that this PLA(2)-like protein obtained during purification of TSH was derived from the endolymphatic sac adhering to the pituitary and that this protein is a bullfrog otoconin. (J Histochem Cytochem 49:631-637, 2001)
Assuntos
Saco Endolinfático/metabolismo , Glicoproteínas/metabolismo , Fosfolipases A/química , Hipófise/metabolismo , Animais , Western Blotting , Proteínas de Ligação ao Cálcio , Saco Endolinfático/ultraestrutura , Feminino , Glicoproteínas/química , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Peso Molecular , Hipófise/ultraestrutura , Rana catesbeiana , Proteínas de XenopusRESUMO
The surface of most cells is covered by glycoconjugates. The composition and thickness of the surface coat varies among different cell types. The purpose of the present study was to demonstrate the presence of and to characterize the cell coat surrounding the cells in the saccular macula of the rainbow trout. Tissues were fixed in Karnovsky's fixative containing either ruthenium red (0.5, 1, or 2%) or tannic acid (1, 2, or 4%). The apical surface of the sensory and supporting cells reacted with both agents. Varying the concentration of the compounds within a certain range did not significantly affect the degree of tissue staining. Whereas ruthenium red staining was distributed evenly along the luminal surface of the epithelium and along the length of the stereocilia, tannic acid formed electron-dense clumps on the luminal surface of sensory and non-sensory cells and in the basal region of the macular epithelium. The stereocilia of the sensory cells also exhibited tannic acid-positive, electrondense precipitate, particularly near the distal ends of these processes, while uniform staining of the plasma membrane was seen along their lengths. The results of this study suggest that the trout saccular macula is provided with extracellular microenvironments which may be necessary for functional integrity.
Assuntos
Saco Endolinfático/citologia , Histocitoquímica/métodos , Taninos Hidrolisáveis , Membrana dos Otólitos/ultraestrutura , Rutênio Vermelho , Truta/anatomia & histologia , Animais , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Saco Endolinfático/metabolismo , Saco Endolinfático/ultraestrutura , Células Epiteliais , Epitélio/metabolismo , Epitélio/ultraestrutura , Microscopia Eletrônica/métodos , Membrana dos Otólitos/metabolismo , Órgãos dos Sentidos/citologia , Órgãos dos Sentidos/metabolismo , Órgãos dos Sentidos/ultraestruturaRESUMO
A systematic study of the effects of acoustic overstimulation on the endolymphatic sac (ES) in the guinea pig was performed. The ES was studied with light and transmission electron microscopy after exposure of the animals to a 3.85 kHz pure tone of 108 dB SPL or 120 dB SPL for 22.5 min (sound energy 9.4 and 150 Pa2 X h, respectively). The damage pattern in the organ of Corti was studied after various post-exposure times with SEM and correlated with the morphological characteristics of the ES in the same ear. This was made possible by using a modified technique for histological processing. In ears with induced structural abnormalities to the organ of Corti, the ES displayed few morphological changes without obvious signs of accumulation of cell debris within the lumen. Initially an increase in the amount of freely floating cells was found which persisted for at least 24 h. The role of the ES for disposal and digestion of locally produced degeneration products within the cochlea after acoustically generated structural damage is discussed.
Assuntos
Estimulação Acústica , Cóclea/ultraestrutura , Orelha Interna/fisiologia , Saco Endolinfático/fisiologia , Estimulação Acústica/métodos , Animais , Audiometria de Tons Puros , Saco Endolinfático/ultraestrutura , Feminino , Cobaias , Células Ciliadas Auditivas/ultraestrutura , Microscopia Eletrônica de Varredura , Órgão Espiral/ultraestruturaRESUMO
It has recently been demonstrated that endolymphatic sac (ES) ribosome-rich (dark) cells respond to induced endolymph changes and are thus likely to be involved in endolymph homeostasis. Therefore, we studied the ultrastructural characteristics of rat ES ribosome-rich cells during development in order to determine the cellular distribution of organelles involved in protein metabolism, secretion and absorption, indicative for their contribution to endolymph homeostasis. During embryonal stages ribosome-rich cells contain a limited number and variety of organelles and are predominantly involved in the production of components for cell growth and differentiation. In the young adult stage (P60) three different states of ribosome-rich cells may be distinguished. State A resembles a cell with only limited metabolic activities whereas state B is characterized by numerous different intracellular organelles and is considered to be involved in production and secretion as well as absorption and degradation of complex proteins. A third cellular state, state C, is filled with phagolysosomes and contains very few other organelles. This is considered to be a final (pre)apoptotic state. Autoradiography data suggest that ES ribosome-rich cells are capable of synthesis and secretion of tyrosine-containing proteins and may thus be involved in regulation of the osmolarity of endolymph based on the capacity to bind cations as well as water molecules. In addition, ES ribosome-rich cells appear to synthesize and secrete fucosylated glycoproteins into the endolymph. In conclusion, the present data suggest that ES ribosome-rich cells are actively involved in endolymph homeostasis through secretion and absorption of complex proteins and it is hypothesized that they are able to adapt their function or activities in response to changes in endolymph composition.
Assuntos
Saco Endolinfático/embriologia , Saco Endolinfático/ultraestrutura , Ribossomos/ultraestrutura , Envelhecimento , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Autorradiografia , Desenvolvimento Embrionário e Fetal , Saco Endolinfático/citologia , Saco Endolinfático/metabolismo , Fucose/farmacocinética , Microscopia Eletrônica , Organelas/metabolismo , Organelas/ultraestrutura , Ratos , Ratos Wistar , Tirosina/farmacocinéticaRESUMO
The presence of separate bundles of nerve fibres in the gerbilline endolymphatic sac (ES) is described, paying particular attention to their ultrastructure and localization. One bundle, localized in the area of the subepithelium which separates the sigmoid sinus from the ES, is composed only of myelinated fascicles which, moreover, seem to have an isolated contact with the ES area. Other two single nerve fibres, much smaller in caliber, are localized in the ES subepithelium and laterally to the ES area, still close to the sigmoid sinus. These fibres, composed of myelinated and unmyelinated fascicles, seem to have a rather longitudinal orientation and, moreover, contract close relationships with the rich vascular network of the ES subepithelial tissue. As far as the course is concerned, the serial sectioning technique would suggest that the nerve fibres get very close to the ES epithelial cell layer, going proximal to distal. Speculations on the origin of this nerve contingent in the ES are proposed and discussed in view of possible new theories for pathogenesis and therapy of some inner ear diseases.
Assuntos
Endolinfa/metabolismo , Saco Endolinfático/inervação , Fibras Nervosas/ultraestrutura , Animais , Doenças Cocleares/etiologia , Doenças Cocleares/fisiopatologia , Modelos Animais de Doenças , Saco Endolinfático/metabolismo , Saco Endolinfático/ultraestrutura , Gerbillinae , Microscopia EletrônicaRESUMO
When the organ of Corti is severely traumatized by intense (160 dB) blast waves, such that as much as 7 mm of Corti's organ is torn loose from the basilar membrane, the cells of the inner sulcus respond to the altered biochemical milieu of the endolymph by a profile elaboration of surface membrane, zeiosis and the development of numerous pseudopodia and microvilli. On the basis of our longitudinal study, this surface reaction appears to peak at approximately 10 days after trauma and may indicate that the inner sulcus cells are extremely active in the endocytosis of cellular debris. Signs of active changes on the surface of the inner sulcus cells occur immediately following trauma, and activity continues for as long as 30 days after exposure. The cells of Claudius, as well as other epithelial cells on the basilar membrane, are also capable of extreme membrane proliferation and mobility. Possible mechanisms for the unusual behavior and the role of the inner sulcus cells in the normal functioning cochlea are discussed.
Assuntos
Orelha Interna/ultraestrutura , Endocitose , Perda Auditiva Provocada por Ruído/patologia , Animais , Membrana Basilar/ultraestrutura , Membrana Celular/ultraestrutura , Chinchila , Saco Endolinfático/ultraestrutura , Epitélio/ultraestrutura , Células Ciliadas Auditivas/ultraestrutura , Microvilosidades/ultraestrutura , Órgão Espiral/ultraestrutura , Fatores de TempoRESUMO
The combined effects of glycerol and colchicine on the endolymphatic sac were investigated in mice. Glycerol induced signs of secretion from the epithelium with formation of secretory granules in the light epithelial cells. Other characteristics of the epithelial lining were also changed resulting in an increased widening of the lateral intercellular spaces, a partial collapse of the lumen and with a deposition of a stainable substance within the lumen. This reaction lasted from 30 min to 24 h following the injection. Pretreatment with colchicine was found to decrease or inhibit the glycerol-induced secretion of macromolecules into the sac. The lumen collapsed but frequently there was no presence of stainable substance. Animals treated with both glycerol and colchicine showed marked signs of inner ear malfunction which could indicate that the secretory activity in the sac might be closely related to the regulation of inner ear fluid homeostasis and that functional disturbances in this system may lead to disorders of inner ear function.
Assuntos
Colchicina/farmacologia , Orelha Interna/metabolismo , Saco Endolinfático/metabolismo , Glicerol/farmacologia , Animais , Grânulos Citoplasmáticos/efeitos dos fármacos , Grânulos Citoplasmáticos/ultraestrutura , Saco Endolinfático/efeitos dos fármacos , Saco Endolinfático/ultraestrutura , Camundongos , Microscopia EletrônicaRESUMO
C-type natriuretic peptide (CNP) is a member of the atrial natriuretic peptide family (ANP family). The family also includes ANP and brain natriuretic peptide (BNP). These peptides regulate the homeostasis of body fluid and blood pressure as a neuropeptide in the central nervous system as well as a cardiac hormone in the periphery. We have recently reported the expression of CNP mRNA in the inner ear. To assess the possible physiological role of CNP in the inner ear, we investigated the localization of CNP peptide in the rat inner ear by immunohistochemistry at the light and electron microscopic level. CNP-like immunoreactivity was widely distributed in the secretory and the neuronal portion of the inner ear, i.e. the spiral ligament, the dark cell region of the utriculus, the epithelium of the endolymphatic sac, the spiral ganglion cells and the vestibular ganglion cells. The results suggest that CNP may play a role in the homeostasis of the perilymph and endolymph and may also influence nerve activities in the inner ear.
Assuntos
Orelha Interna/metabolismo , Peptídeo Natriurético Tipo C/metabolismo , Animais , Cóclea/metabolismo , Cóclea/ultraestrutura , Orelha Interna/ultraestrutura , Saco Endolinfático/metabolismo , Saco Endolinfático/ultraestrutura , Imuno-Histoquímica , Masculino , Microscopia Imunoeletrônica , Ratos , Ratos Wistar , Distribuição Tecidual , Vestíbulo do Labirinto/metabolismo , Vestíbulo do Labirinto/ultraestruturaRESUMO
Morphological changes of the endolymphatic sac were analyzed in guinea pigs following microinjection of artificial endolymph into the cochlea or withdrawal of a quantity of native endolymph. Injections were performed into the second turn of scala media with a micro-pump at a rate of 60-100 nl/min, lasting for a period of 4, 7. 5, 15 or 18 min. In withdrawal experiments, endolymph was aspirated from the second cochlear turn over a period of 8 min. For each procedure the contralateral (non-treated) ear served as a histological control. Following artificial endolymph injections of 7. 5 min or more there was an almost total absence of the normal intraluminal homogeneous substance (HS) on the injected side. Our observations suggest that the disappearance of the HS occurs by both enzymatic and macrophagic activity. After endolymphatic withdrawals the ES was found to contain increased amounts of HS. The results could suggest that the volume of fluid in the ES, and hence the volume of the entire membranous labyrinth, may be regulated by a dynamic relationship between active secretion and enzymatic degradation of a lumen-expanding substance that is intimately related to the intraluminal macrophages. The exact mechanism governing these regulatory systems, and their relationship to ion and water movements across the epithelium of the sac, remain to be elucidated.
Assuntos
Cóclea/efeitos dos fármacos , Endolinfa/fisiologia , Saco Endolinfático/ultraestrutura , Animais , Cóclea/fisiologia , Eletrofisiologia , Endolinfa/química , Ducto Endolinfático/ultraestrutura , Saco Endolinfático/química , Saco Endolinfático/efeitos dos fármacos , Cobaias , Microinjeções , Microscopia EletrônicaRESUMO
The effects of hyperosmolar substances on the ultrastructure of the endolymphatic sac were studied in mice. Fifteen minutes after intravenous injection of urea or glycerol, subcellular changes in the endolymphatic sac were observed. These consisted of the occurrence of abundant cytoplasmic granules with a floccular or lamellar material, or both, in the light epithelial cells. Similar material was also present in the lumen of the endolymphatic sac, suggesting a common source and increased secretory activity. Mannitol caused similar changes, though less pronounced. The possibility that the alterations in the fine structure of the endolymphatic sac may be associated with a reduction in the hydrostatic fluid pressure in the rest of the labyrinth is discussed.
Assuntos
Orelha Interna/efeitos dos fármacos , Saco Endolinfático/efeitos dos fármacos , Glicerol/farmacologia , Manitol/farmacologia , Ureia/farmacologia , Animais , Grânulos Citoplasmáticos/efeitos dos fármacos , Grânulos Citoplasmáticos/ultraestrutura , Endolinfa/efeitos dos fármacos , Saco Endolinfático/ultraestrutura , Epitélio/efeitos dos fármacos , Epitélio/ultraestrutura , Feminino , Masculino , Camundongos , Camundongos EndogâmicosRESUMO
The effects of glycerol and colchicine on the sensory end organs of the inner ear were investigated in mice. Glycerol alone induced a widening of the intercellular spaces lining vestibular dark and transitional cells as well as the marginal cells of the stria vascularis. This was noted within 30 min after the injection of glycerol and was normalized again within 4 h after the injection. Colchicine induced some morphological changes in the inner ear sensory cells, such as dissociation of Golgi complexes etc. These isolated glycerol or colchicine injections did not cause any signs of inner ear functional impairment. Treatment with glycerol following pretreatment with colchicine, however, induced marked inner ear dysfunction with impaired sense of balance and audition. The inner ear morphology revealed a combination of changes as compared with what was observed after isolated treatment with glycerol or colchicine i.e. edema of the stria vascularis, and vestibular dark and transitional cells as well as dissociation of Golgi complexes in the sensory cells. The cochlea showed moderate endolymphatic hydrops. These findings indicate that colchicine affects the inner ear fluid regulating mechanisms which may lead to severe functional derangement after additional glycerol treatment. It is conceivable that the present experiment may serve as a useful model for further studies on inner ear changes related to endolymphatic hydrops and Ménière's disease.