Entwicklung textiler Maschinenelemente für den Einsatz in Windkraftanlagen und Seilwinden

Es sollen hochfeste, gewichtreduzierte Zug- und Tragmittel aus hochmodularen (HM) und hochfesten (HT) Fasern validiert und dabei sowohl runde als auch flache, riemenartige Strukturen untersucht werden. Dadurch sind effizientere Fördersysteme und die Überwindung technischer Grenzen möglich. Darüber hinaus soll das Hauptkriterium für ein breites Anwendungsspektrum geschaffen werden: ein anerkanntes, zerstörungsfreies Prüfverfahren, mit dem der Austausch- bzw. Wartungszeitpunkt des textilen Tragmittels bestimmt werden kann. Können die o. g. Punkte erfolgreich bearbeitet werden, erfolgt eine Ausdehnung der textilen Strukturen in den Bereich kraftübertragender Maschinenelemente. Anhand von Feldversuchen in fördertechnischen Anlagen im Bergbau/ Intralogistik soll erstmals der vollständige Nachweis geführt werden, dass derartige textile Strukturen in technischen Anwendungen eingesetzt werden können. Der Nachweis umfasst die Validierung einer Vielzahl von Einzelschwerpunkten wie die Entwicklung einer Endlos-Herstellungstechnologie bzw. Endverbindung, die Tragmitteldimensionierung, die Erbringung von Festigkeitsnachweisen, die Erarbeitung von Vorschriften und die Erprobung der Verfahren zur Zustandsüberwachung.

DEFINITION OF THE LANDSCAPE OF CHROMATIN STRUCTURE AT THE FRATAXIN GENE IN FRIEDREICH’S ATAXIA

Friedreich’s ataxia (FRDA) is caused by the transcriptional silencing of the frataxin (FXN) gene. FRDA patients have expansion of GAA repeats in intron 1 of the FXN gene in both alleles. A number of studies demonstrated that specific histone deacetylase inhibitors (HDACi) affect either histone modifications at the FXN gene or FXN expression in FRDA cells, indicating that the hyperexpanded GAA repeat may facilitate heterochromatin formation. However, the correlation between chromatin structure and transcription at the FXN gene is currently limited due to a lack of more detailed analysis. Therefore, I analyzed the effects of the hyperexpanded GAA repeats on transcription status and chromatin structure using lymphoid cell lines derived from FRDA patients. Using chromatin immunoprecipitation and quantitative PCR, I observed significant changes in the landscape of histone modifications in the vicinity of the GAA tract in FRDA cells relative to control cells. Similar epigenetic changes were observed in GFP reporter construct containing 560 GAA repeats. Further, I detected similar levels of FXN pre-mRNA at a region upstream of hyperexpanded GAA repeats in FRDA and control cells, indicating similar efficiency of transcription initiation in FRDA cells. I also showed that histone modifications associated with hyperexpanded GAA repeats are independent of transcription progression using the GFP reporter system. My data strongly support evidence that FXN deficiency in FRDA patients is consequence of defective transition from initiation to elongation of FXN transcription due to heterochromatin-like structures formed in the proximity of the hyperexpanded GAAs.

Paracrine or virus-mediated induction of decorin expression by endothelial cells contributes to tube formation and prevention of apoptosis in collagen lattices

Resting endothelial cells express the small proteoglycan biglycan, whereas sprouting endothelial cells also synthesize decorin, a related proteoglycan. Here we show that decorin is expressed in endothelial cells in human granulomatous tissue. For in vitro investigations, the human endothelium-derived cell line, EA.hy 926, was cultured for 6 or more days in the presence of 1% fetal calf serum on top of or within floating collagen lattices which were also populated by a small number of rat fibroblasts. Endothelial cells aligned in cord-like structures and developed cavities that were surrounded by human decorin. About 14% and 20% of endothelial cells became apoptotic after 6 and 12 days of co-culture, respectively. In the absence of fibroblasts, however, the extent of apoptosis was about 60% after 12 days, and cord-like structures were not formed nor could decorin production be induced. This was also the case when lattices populated by EA.hy 926 cells were maintained under one of the following conditions: 1) 10% fetal calf serum; 2) fibroblast-conditioned media; 3) exogenous decorin; or 4) treatment with individual growth factors known to be involved in angiogenesis. The mechanism(s) by which fibroblasts induce an angiogenic phenotype in EA.hy 926 cells is (are) not known, but a causal relationship between decorin expression and endothelial cell phenotype was suggested by transducing human decorin cDNA into EA.hy 926 cells using a replication-deficient adenovirus. When the transduced cells were cultured in collagen lattices, there was no requirement of fibroblasts for the formation of capillary-like structures and apoptosis was reduced. Thus, decorin expression seems to be of special importance for the survival of EA.hy 926 cells as well as for cord and tube formation in this angiogenesis model.

Colocalization of a large heterodimeric proteoglycan with basement membrane proteins in cultured cells.

A novel large heterodimeric dermatan sulfate proteoglycan with core proteins of 460 and 300 kDa, respectively, had been described as a secretory product of human fetal skin fibroblasts (Breuer et al., J. Biol. Chem. 266, 13224-13232 (1991)). Pulse-chase experiments showed a preferential association of the proteoglycan with the cell membrane. Immunogold labeling indicated its localization in fibrils on the cell surface as well as in fibrillar extensions from the cell body. Immunofluorescence studies yielded a fibrillar and punctate staining pattern which was also seen in cultured human and porcine endothelial cells. Dot-like structures were observed in transformed human keratinocytes. Various immunocytochemical double-labeling experiments indicated a remarkable colocalization of the proteoglycan with fibronectin, laminin, perlecan, and type IV collagen whereas only occasionally a colocalization with chondroitin-6-sulfate was found. No evidence for an enrichment of the proteoglycan in vinculin-containing structures was obtained. These results suggest that the proteoglycan is a widely distributed macromolecule which can associate with basement membrane components. Preliminary findings in rat cornea supported this conclusion.

A growth factor-induced, spatially organizing cytoskeletal module enables rapid and persistent fibroblast migration

Directional migration requires robust front/back polarity. We find that fibroblasts treated with platelet-derived growth factor (PDGF) and prepolarized by plating on a fibronectin line substrate exhibit persistent migration for hours. This does not occur in the absence of PDGF or on uniformly coated fibronectin substrates. Persistent migration arises from establishment of two functional modules at cell front and back. At the front, formation of a zone containing podosome-like structures (PLS) dynamically correlates with low RhoA and myosin activity and absence of a contractile lamella. At the back, myosin contractility specifically controls tail retraction with minimal crosstalk to the front module. The PLS zone is maintained in a dynamic steady state that preserves size and position relative to the cell front, allowing for long-term coordination of front and back modules. We propose that front/back uncoupling achieved by the PLS zone is crucial for persistent migration in the absence of directional cues.

Conditional expression of Sry, and Wnt4 by gene-targeting: Studies in sexual and skeletal development

Sry and Wnt4 cDNAs were individually introduced into the ubiquitously-expressed Rosa26 ( R26) locus by gene targeting in embryonic stem (ES) cells to create a conditional gene expression system in mice. In the targeted alleles, expression of these cDNAs should be blocked by a neomycin resistance selection cassette that is flanked by loxP sites. Transgene expression should be activated after the blocking cassette is deleted by Cre recombinase. ^ To test this conditional expression system, I have bred R26-stop- Sry and R26-stop-Wnt4 heterozygotes with a MisRII-Cre mouse line that expresses Cre in the gonads of both sexes. Analysis of these two types of bigenic heterozygotes indicated that their gonads developed normally like those of wild types. However, one XX R26-Sry/R26-Sry; MisR2-Cre/+ showed epididymis-like structures resembling those of males. In contrast, only normal phenotypes were observed in XY R26-Wnt4/R26-Wnt4; MisR2-Cre /+ mice. To interpret these results, I have tested for Cre recombinase activity by Southern blot and transcription of the Sry and Wnt4 transgenes by RT-PCR. Results showed that bigenic mutants had insufficient activation of the transgenes in their gonads at E12.5 and E13.5. Therefore, the failure to observe mutant phenotypes may have resulted from low activity of MisR2-Cre recombination at the appropriate time. ^ Col2a1-Cre transgenic mice express Cre in differentiating chondrocytes. R26-Wnt4; Col2a1-Cre bigenic heterozygous mice were found to exhibit a dramatic alteration in growth presumably caused by Wnt4 overexpression during chondrogenesis. R26-Wnt4; Col2a1-Cre mice exhibited dwarfism beginning approximately 10 days after birth. In addition, they also had craniofacial abnormalities, and had delayed ossification of the lumbar vertebrate and pelvic bones. Histological analysis of the growth plates of R26-Wnt4; Col2a1-Cre mice revealed less structural organization and a delay in onset of the primary and secondary ossification centers. Molecular studies confirmed that overexpression of Wnt4 causes decreased proliferation and early maturation of chondrocytes. In addition, R26-Wnt4; Col2a1-Cre mice had decreased expression of vascular endothelial growth factor (VEGF), suggesting that defects in vascularization may contribute to the dwarf phenotype. Finally, 9-month-old R26-Wnt4; Col2a1-Cre mice had significantly more fat cells in the marrow cavities of their metaphysis long bones, implying that long-term overexpression of Wnt4may cause bone marrow pathologies. In conclusion, Wnt4 was activated by Col2a1-Cre recombinase and was overexpressed in the growth plate, resulting in aberrant proliferation and differentiation of chondrocytes, and ultimately leads to dwarfism in mice. ^

Mineralogical and stable isotope analyses of carbonates from the eastern Black Sea

Authigenic carbonates associated with cold seeps provide valuable archives of changes in the long-term seepage activity. To investigate the role of shallow-buried hydrates on the seepage strength and fluid composition we analysed methane-derived carbonate precipitates from a high-flux hydrocarbon seepage area ("Batumi seep area") located on the south-eastern Black Sea slope in ca. 850 m. In a novel approach, we combined computerized X-ray tomography (CT) with mineralogical and isotope geochemical methods to get additional insights into the three-dimensional internal structure of the carbonate build-ups. X-ray diffractometry revealed the presence of two different authigenic carbonate phases, i.e. pure aragonitic rims associated with vital microbial mats and high-Mg calcite cementing the hemipelagic sediment. As indicated by the CT images, the initial sediment has been strongly deformed, first plastic then brittle, leading to brecciation of the progressively cemented sediment. The aragonitic rims on the other hand, represent a presumably recent carbonate growth phase since they cover the already deformed sediment. The stable oxygen isotope signature indicates that the high-Mg calcite cement incorporated pore water mixed with substantial hydrate water amounts. This points at a dominant role of high gas/fluid flux from decomposing gas hydrates leading to the deformation and cementation of the overlying sediment. In contrast, the aragonitic rims do not show an influence of 18O-enriched hydrate water. The differences in d18O between the presumably recent aragonite precipitates and the older high-Mg cements suggest that periods of hydrate dissociation and vigorous fluid discharge alternated with times of hydrate stability and moderate fluid flow. These results indicate that shallow-buried gas hydrates are prone to episodic decomposition with associated vigorous fluid flow. This might have a profound impact on the seafloor morphology resulting e.g. in the formation of carbonate pavements and pockmark-like structures but might also affect the local carbon cycle.

A computer program for the analysis of the dynamic bending-torsion coupling in bridges using a mini-computer

The analysis of modes and natural frequencies is of primary interest in the computation of the response of bridges. In this article the transfer matrix method is applied to this problem to provide a computer code to calculate the natural frequencies and modes of bridge-like structures. The Fortran computer code is suitable for running on small computers and results are presented for a railway bridge.

Hexagons and spiral defect chaos in non-Boussinesq convection at low Prandtl numbers

We study the stability and dynamics of non-Boussinesq convection in pure gases ?CO2 and SF6? with Prandtl numbers near Pr? 1 and in a H2-Xe mixture with Pr= 0.17. Focusing on the strongly nonlinear regime we employ Galerkin stability analyses and direct numerical simulations of the Navier-Stokes equations. For Pr ? 1 and intermediate non-Boussinesq effects we find reentrance of stable hexagons as the Rayleigh number is increased. For stronger non-Boussinesq effects the usual, transverse side-band instability is superseded by a longitudinal side-band instability. Moreover, the hexagons do not exhibit any amplitude instability to rolls. Seemingly, this result contradicts the experimentally observed transition from hexagons to rolls. We resolve this discrepancy by including the effect of the lateral walls. Non-Boussinesq effects modify the spiral defect chaos observed for larger Rayleigh numbers. For convection in SF6 we find that non-Boussinesq effects strongly increase the number of small, compact convection cells and with it enhance the cellular character of the patterns. In H2-Xe, closer to threshold, we find instead an enhanced tendency toward roll-like structures. In both cases the number of spirals and of targetlike components is reduced. We quantify these effects using recently developed diagnostics of the geometric properties of the patterns.

Investigación experimental del flujo aerodinámico sobre helipuertos embarcados

La investigación del flujo aerodinámico sobre helipuertos embarcados se encuentra estrechamente relacionada con la operación segura de las aeronaves, pues las condiciones del flujo que tiene lugar en ese entorno pueden exceder los límites para los que están certificadas dichas aeronaves. El ambiente aerodinámico en las inmediaciones de un barco es altamente complejo y se encuentra influenciado por gran número de factores (chimeneas, antenas, mástiles, etc.) relacionados con la configuración específica del propio barco. El flujo objeto de investigación corresponde a la estela que se desarrolla sobre la cubierta de vuelo de una fragata, el cual está fuertemente influenciado por la superestructura de la misma, y que cualitativamente es similar al flujo que tiene lugar entre edificios altos o helipuertos situados en áreas urbanas, pues comprende estructuras tipo caja, con bordes afilados, que generan flujos tridimensionales altamente turbulentos. En esta Tesis se aborda el estudio del problema desde el punto de vista experimental, mediante simulación en túnel aerodinámico y medida de las variables del campo fluido sobre maquetas de fragatas a escala reducida. Las herramientas empleadas para tal cometido, han sido técnicas experimentales, tales como la visualización del flujo, la velocimetría láser por imágenes de partículas, la anemometría láser Doppler y los scanners electrónicos de presión, que han permitido investigar el flujo problema con objeto de obtener información, y adquirir así, un conocimiento más profundo de dicho flujo. La explotación de este conocimiento, ha dado lugar al diseño de una nueva solución, basada en la modificación de geometría básica de la fragata, por medio del cambio de la curvatura del techo del hangar, permitiendo suavizar el escalón descendente que se produce aguas abajo del mismo. Las geometrías modificadas han sido ensayada en túnel mediante la misma metodología empleada para la fragata original, de modo que, ha podido establecerse un análisis comparativo, para valorar la efectividad de la solución propuesta, el cual ha mostrado resultados satisfactorios, retirando el flujo adverso de la zona de operación de helicópteros y desplazándolo hacia el hangar, donde resulta menos peligroso, de modo que se reduce la carga del piloto y los riesgos de accidente durante las operaciones a bordo de embarcaciones. ABSTRACT The investigation of aerodynamic flow above the ship’s heliports is directly related to the aircraft safe operation, because the environment flow conditions may exceed the aircraft certification limits. Aerodynamic ship’s environment is highly complex and it is influenced by a large number of factors (stacks, antennae, masts, …) related to each specific ship configuration. The flow under investigation occurs into the wake produced above the flight deck of a frigate, that is strongly influenced by the superstructure. This flow is similar to one produced around tall buildings or heliports located in urban areas, thus in both of them, the air is flowing around sharp-edges box-like structures, producing three-dimensional and highly turbulent flows. This Thesis studies the problem from an experimental point of view, by means of wind tunnel simulations and measurements of the flow field around reduced scale frigates models. Tools used in this work are the experimental techniques, as flow visualization, particle image velocimetry, laser Doppler anemometry and pressure electronic scanners. These techniques provide information about the flow in order to obtain a more complete insight of this kind of flows. The exploitation of this insight is used for the design of a new flow control concept, based on the modification of the basic frigate geometry. This new design consists in the hangar roof curvature modification that produces a smoothing of the descendent step located downstream the hangar. Modified geometries are tested in wind tunnel by means of the same methodology as the original frigate, thus a comparative analysis is established in order to perform an assessment of effectiveness. This analysis has shown good results in displacing the adverse flow from the helicopter operation path to the nearest hangar region, reducing the pilot load and the accident risks during on board operations.

Distributed, layered and reliable computing nets to represent neuronal receptive fields

Abstract. Receptive fields of retinal and other sensory neurons show a large variety of spatiotemporal linear and non linear types of responses to local stimuli. In visual neurons, these responses present either asymmetric sensitive zones or center-surround organization. In most cases, the nature of the responses suggests the existence of a kind of distributed computation prior to the integration by the final cell which is evidently supported by the anatomy. We describe a new kind of discrete and continuous filters to model the kind of computations taking place in the receptive fields of retinal cells. To show their performance in the analysis of diferent non-trivial neuron-like structures, we use a computer tool specifically programmed by the authors to that efect. This tool is also extended to study the efect of lesions on the whole performance of our model nets.

Structural efficiency variation with the problem size in some bending problems = Variación del rendimiento estructural con el tamaño del problema en algunos casos de flexión simple

Existe normalmente el propósito de obtener la mejor solución posible cuando se plantea un problema estructural, entendiendo como mejor la solución que cumpliendo los requisitos estructurales, de uso, etc., tiene un coste físico menor. En una primera aproximación se puede representar el coste físico por medio del peso propio de la estructura, lo que permite plantear la búsqueda de la mejor solución como la de menor peso. Desde un punto de vista práctico, la obtención de buenas soluciones—es decir, soluciones cuyo coste sea solo ligeramente mayor que el de la mejor solución— es una tarea tan importante como la obtención de óptimos absolutos, algo en general difícilmente abordable. Para disponer de una medida de la eficiencia que haga posible la comparación entre soluciones se propone la siguiente definición de rendimiento estructural: la razón entre la carga útil que hay que soportar y la carga total que hay que contabilizar (la suma de la carga útil y el peso propio). La forma estructural puede considerarse compuesta por cuatro conceptos, que junto con el material, definen una estructura: tamaño, esquema, proporción, y grueso.Galileo (1638) propuso la existencia de un tamaño insuperable para cada problema estructural— el tamaño para el que el peso propio agota una estructura para un esquema y proporción dados—. Dicho tamaño, o alcance estructural, será distinto para cada material utilizado; la única información necesaria del material para su determinación es la razón entre su resistencia y su peso especifico, una magnitud a la que denominamos alcance del material. En estructuras de tamaño muy pequeño en relación con su alcance estructural la anterior definición de rendimiento es inútil. En este caso —estructuras de “talla nula” en las que el peso propio es despreciable frente a la carga útil— se propone como medida del coste la magnitud adimensional que denominamos número de Michell, que se deriva de la “cantidad” introducida por A. G. M. Michell en su artículo seminal de 1904, desarrollado a partir de un lema de J. C. Maxwell de 1870. A finales del siglo pasado, R. Aroca combino las teorías de Galileo y de Maxwell y Michell, proponiendo una regla de diseño de fácil aplicación (regla GA), que permite la estimación del alcance y del rendimiento de una forma estructural. En el presente trabajo se estudia la eficiencia de estructuras trianguladas en problemas estructurales de flexión, teniendo en cuenta la influencia del tamaño. Por un lado, en el caso de estructuras de tamaño nulo se exploran esquemas cercanos al optimo mediante diversos métodos de minoración, con el objetivo de obtener formas cuyo coste (medido con su numero deMichell) sea muy próximo al del optimo absoluto pero obteniendo una reducción importante de su complejidad. Por otro lado, se presenta un método para determinar el alcance estructural de estructuras trianguladas (teniendo en cuenta el efecto local de las flexiones en los elementos de dichas estructuras), comparando su resultado con el obtenido al aplicar la regla GA, mostrando las condiciones en las que es de aplicación. Por último se identifican las líneas de investigación futura: la medida de la complejidad; la contabilidad del coste de las cimentaciones y la extensión de los métodos de minoración cuando se tiene en cuenta el peso propio. ABSTRACT When a structural problem is posed, the intention is usually to obtain the best solution, understanding this as the solution that fulfilling the different requirements: structural, use, etc., has the lowest physical cost. In a first approximation, the physical cost can be represented by the self-weight of the structure; this allows to consider the search of the best solution as the one with the lowest self-weight. But, from a practical point of view, obtaining good solutions—i.e. solutions with higher although comparable physical cost than the optimum— can be as important as finding the optimal ones, because this is, generally, a not affordable task. In order to have a measure of the efficiency that allows the comparison between different solutions, a definition of structural efficiency is proposed: the ratio between the useful load and the total load —i.e. the useful load plus the self-weight resulting of the structural sizing—. The structural form can be considered to be formed by four concepts, which together with its material, completely define a particular structure. These are: Size, Schema, Slenderness or Proportion, and Thickness. Galileo (1638) postulated the existence of an insurmountable size for structural problems—the size for which a structure with a given schema and a given slenderness, is only able to resist its self-weight—. Such size, or structural scope will be different for every different used material; the only needed information about the material to determine such size is the ratio between its allowable stress and its specific weight: a characteristic length that we name material structural scope. The definition of efficiency given above is not useful for structures that have a small size in comparison with the insurmountable size. In this case—structures with null size, inwhich the self-weight is negligible in comparisonwith the useful load—we use as measure of the cost the dimensionless magnitude that we call Michell’s number, an amount derived from the “quantity” introduced by A. G. M. Michell in his seminal article published in 1904, developed out of a result from J. C.Maxwell of 1870. R. Aroca joined the theories of Galileo and the theories of Maxwell and Michell, obtaining some design rules of direct application (that we denominate “GA rule”), that allow the estimation of the structural scope and the efficiency of a structural schema. In this work the efficiency of truss-like structures resolving bending problems is studied, taking into consideration the influence of the size. On the one hand, in the case of structures with null size, near-optimal layouts are explored using several minimization methods, in order to obtain forms with cost near to the absolute optimum but with a significant reduction of the complexity. On the other hand, a method for the determination of the insurmountable size for truss-like structures is shown, having into account local bending effects. The results are checked with the GA rule, showing the conditions in which it is applicable. Finally, some directions for future research are proposed: the measure of the complexity, the cost of foundations and the extension of optimization methods having into account the self-weight.

mRNA binding protein mrnp 41 localizes to both nucleus and cytoplasm

We have identified and molecularly characterized a human protein with a Mr of 40,880 Da. After UV irradiation of HeLa cells, this protein was cross-linked to poly(A)-containing mRNA and was therefore designated mrnp 41 (for mRNA binding protein of 41 kDa). Cell fractionation and immunoblotting showed mrnp 41 in both the cytoplasm and the nucleus and particularly in the nuclear envelope. Immunofluorescence microscopy localized mrnp 41 to distinct foci in the nucleoplasm, to the nuclear rim, and to meshwork-like structures throughout the cytoplasm. The cytoplasmic meshwork staining was disrupted by prior treatment of cells with the actin filament- or microtubule-disrupting drugs cytochalasin or nocodazole, respectively, suggesting association of mrnp 41 with the cytoskeleton. Double immunofluorescence with antibodies against mrnp 41 and the cytoplasmic poly(A) binding protein showed colocalization to the cytoplasmic meshwork. Immunogold electronmicroscopy confirmed mrnp 41’s cytoplasmic and nucleoplasmic localization and revealed a striking labeling of nuclear pore complexes. Together these data suggest that mrnp 41 may function in nuclear export of mRNPs and/or in cytoplasmic transport on, or attachment to, the cytoskeleton. Consistent with a role of mrnp 41 in nuclear export are previous reports that mutations in homologs of mrnp 41 in Schizosaccharomyces pombe, designated Rae1p, or in Saccharomyces cerevisiae, designated Gle2p, result in mRNA accumulation in the nucleus although it is presently not known whether these homologs are mRNA binding proteins as well.

Axon pathology in Parkinson’s disease and Lewy body dementia hippocampus contains α-, β-, and γ-synuclein

Pathogenic α-synuclein (αS) gene mutations occur in rare familial Parkinson’s disease (PD) kindreds, and wild-type αS is a major component of Lewy bodies (LBs) in sporadic PD, dementia with LBs (DLB), and the LB variant of Alzheimer’s disease, but β-synuclein (βS) and γ-synuclein (γS) have not yet been implicated in neurological disorders. Here we show that in PD and DLB, but not normal brains, antibodies to αS and βS reveal novel presynaptic axon terminal pathology in the hippocampal dentate, hilar, and CA2/3 regions, whereas antibodies to γS detect previously unrecognized axonal spheroid-like lesions in the hippocampal dentate molecular layer. The aggregation of other synaptic proteins and synaptic vesicle-like structures in the αS- and βS-labeled hilar dystrophic neurites suggests that synaptic dysfunction may result from these lesions. Our findings broaden the concept of neurodegenerative “synucleinopathies” by implicating βS and γS, in addition to αS, in the onset/progression of PD and DLB.

Otogelin: A glycoprotein specific to the acellular membranes of the inner ear

Efforts to identify the specific components of the mammalian inner ear have been hampered by the small number of neuroepithelial cells and the variety of supporting cells. To circumvent these difficulties, we used a PCR-based subtractive method on cDNA from 2-day-old mouse cochlea. A cDNA encoding a predicted 2910-amino acid protein related to mucin has been isolated. Several lines of evidence indicate, however, that this protein does not undergo the O-glycosylation characteristic to mucins. As confirmed by immunocytochemistry and biochemical experiments, this protein is specific to the inner ear. Immunohistofluorescence labeling showed that this protein is a component of all the acellular membranes of the inner ear: i.e., the tectorial membrane of the cochlea, the otoconial and accessory membranes of the utricule and saccule, the cupula of the semicircular canals, and a previously undescribed acellular material covering the otoconia of the saccule. The protein has been named otogelin with reference to its localization. A variety of nonsensory cells located underneath these membranes could be identified as synthesizing otogelin. Finally, this study revealed a maturation process of the tectorial membrane, as evidenced by the progressive organization of otogelin labeling into thick and spaced radial fiber-like structures.