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Tight junctions form a morphological and physical border between the apical and the basolateral cell surface domains of epithelial cells; hence assembly of tight junctions could occur from both of the two plasma membrane domains. We show here that the C-terminal cytoplasmic domain of occludin, the only known transmembrane protein of tight junctions, was sufficient to mediate basolateral expression of a chimeric protein. Since this chimera was transported directly to the basolateral membrane during biosynthesis, the C-terminal domain of occludin contains a basolateral targeting signal. Additionally, the C-terminal domain of occludin was also able to mediate endocytosis. Thus, the C-terminal cytoplasmic domain appears to govern intracellular transport of occludin. To test whether the basolateral membrane is an obligatory intermediate in transport of occludin to tight junctions, we analyzed the expression of occludin molecules rendered unable to efficiently integrate into tight junctions by the introduction of N-linked glycosylation sites into the two extracellular loops. Indeed, glycosylated occludin accumulated in the basolateral membrane, supporting a model in which the biogenesis of tight junctions occurs from this cell-surface domain.

Cell-cell adhesion regulates the development and function of epithelia by providing mechanical support and by guiding cell proliferation and differentiation. The tight junction (TJ) protein zonula occludens (ZO)-1 regulates cell proliferation and gene expression by inhibiting the activity of the Y-box transcription factor ZONAB in cultured epithelial cells. We investigated the role of this TJ-associated signalling pathway in the retinal pigment epithelium (RPE) in vivo by lentivirally-mediated overexpression of ZONAB, and knockdown of its cellular inhibitor ZO-1. Both overexpression of ZONAB or knockdown of ZO-1 resulted in increased RPE proliferation, and induced ultrastructural changes of an epithelial-mesenchymal transition (EMT)-like phenotype. Electron microscopy analysis revealed that transduced RPE monolayers were disorganised with increased pyknosis and monolayer breaks, correlating with increased expression of several EMT markers. Moreover, fluorescein angiography analysis demonstrated that the increased proliferation and EMT-like phenotype induced by overexpression of ZONAB or downregulation of ZO-1 resulted in RPE dysfunction. These findings demonstrate that ZO-1 and ZONAB are critical for differentiation and homeostasis of the RPE monolayer and may be involved in RPE disorders such as proliferative vitroretinopathy and atrophic age-related macular degeneration.

Epithelial tight junctions recruit different types of signalling proteins that regulate cell proliferation and differentiation. Little is known about how such proteins interact functionally and biochemically with each other. Here, we focus on the Y-box transcription factor ZONAB (zonula occludens 1-associated nucleic-acid-binding protein)/DbpA (DNA-binding protein A) and the Rho GTPase activator guanine nucleotide exchange factor (GEF)-H1/Lbc's first cousin, which are two tight-junction-associated signalling proteins that regulate proliferation. Our data show that the two proteins interact and that ZONAB activity is Rho-dependent. Overexpression of GEF-H1 induces accumulation of ZONAB in the nucleus and activates transcription. Microtubule-affinity regulating kinase/partition-defective-1, another type of GEF-H1-associated signalling protein, remains in the cytoplasm and partially co-localizes with the exchange factor. GEF-H1 and ZONAB are required for expression of endogenous cyclin D1, a crucial RhoA signalling target gene, and GEF-H1-stimulated cyclin D1 promoter activity requires ZONAB. Our data thus indicate that GEF-H1 and ZONAB form a signalling module that mediates Rho-regulated cyclin D1 promoter activation and expression.

[Resumen] En este trabajo se presenta una cartografía detallada de un sistema de cizalla que incluye la zona de cizalla de Villalcampo propiamente dicha y las bandas asociadas. Este sistema tiene carácter regional, afecta a granitos intruidos después de la segunda fase hercínica y a sus encajantes metamórficos en un área de al menos 150 Km2. El estudio geométrico y cinemático de las bandas, la distribución de las rocas de falla, así como el estudio microestructural han permitido interpretar todo el sistema como una cizalla subvenical dextra de carácter dúctil-frágil, que termina hacia el NW en un abanico extensional y que se prolonga hacia el SE en otra amplia área interpretada como un duplex extensional. El valor de la cizalla 'Y es de 1,55 y el desplazamiento mínimo calculado es de 3,7 Km. Todas las bandas de cizalla que incluye el sistema de Villalcampo y las venas o filones asociados pueden relacionarse con un único campo de esfuerzos en el que la trayectoria del esfuerzo principal al, subhorizontal y de dirección aproximadamente N-S sufre una desviación en la zona terminal de la banda en sentido diferente según el labio de que se trate, tal como propone Anderson (1951). Para caracterizar el régimen y los mecanismos de la deformación se ha realizado un análisis microestructural detallado de las rocas de falla y un estudio petrográfico y de la petrofábrica de las milonitas. Se concluye que los yacimientos de oro de Pino están controlados estructuralmente por este sistema de cizalla ya que aparecen en venas extensionales ocupadas por milonitas brechificadas; las condiciones de la deformación de carácter alternativamente dúctil-frágil habrtan permitido la migración de fluidos mineralizadores y el sellado de las fracturas en ciclos repetidos. Se propone como hipótesis que los yacimientos de estaño y wolframio que aparecen en la región de Villaseco-Almaraz de Duero puedan estar controlados también por el duplex extensional de este sistema de cizalla. [Abstract] The present work reports" on a detailed mapping of the Villalcampo shear system, including the Villalcampo shear itself together with related shears and veins. This regional shear system affects granites intruded after the second hercynian deformation phase and their metamorphic host rocks over an area of about 150 Km2. A st~ctural study of the shear bands, encompassing geometric and kinematic criteria and also the distribution of fault rocks, has allowed the authors to interpret the whole of the system as a ductile-fragile subvenical dextral shear spaying to the NW in an extensional fan that is prolonged to the SE over a broad area interpreted as an extensional duplex. The value of the shear strain, 'Y = 1.55, and the minimum displacement, s = 3.700 m, is calculated. The distribution of shear bands and veins is coherent with the notion of a single stress field where the principal stress, o"¡, is subhorizontal and has a N-S trajectory that deviates to the tips of the main shear, as proposed by Anderson (1951). With a view to gaining insight into the shearing regimen and the physical conditions of the deformation, a microestructural exploration of the fault rocks was carried out together with a petrographical and petrofabric study of the mylonites. It is concluded that the Pino ore veins -with Au mineralization- are structurally controlled by this shear system because they are related to the extensional veins infilled by brecchiated mylonites. Additionally, it is proposed that the deformation conditions would have alternated between brittle and ductile; this would have permited the migration and sealing of microfractures by mineralizing fluids during repeated cycles. As an hypothesis, it is proposed that the Sn and W ores situated in -the Villaseco -Almaraz area are possibly related to the extensional duplex.

Tight junctions are the most apical intercellular junctions of epithelial and endothelial cells and create a regulatable semipermeable diffusion barrier between individual cells. On a cellular level, they form an intramembrane diffusion fence that restricts the intermixing of apical and basolateral membrane components. In addition to these well defined functions, more recent evidence suggests that tight junctions are also involved in basic cellular processes like the regulation of cell growth and differentiation.

Muchos de los estudios realizados en los últimos años indican que la sensación de invisibilidad y discriminación que sentimos las mujeres dedicadas a la ciencia, y específicamente a la Geología, no es una mera percepción subjetiva, sino que las cifras están ahí corroborando estas sensaciones. Los datos de distintos organismos y elaborados por distintas fuentes siempre indican una menor participación de las mujeres en los órganos de gestión, y una mayor dificultad para alcanzar los puestos de trabajo de niveles más altos. Esto es así a pesar de que el nº de licenciadas de ciencias supera al de los licenciados, pero a lo largo de la carrera profesional se produce una inversión que hace que las geólogas “se pierdan” por el camino. En este trabajo se presentan algunas de estas cifras con objeto de llamar la atención sobre la situación y discutir en la Mesa Redonda sobre las posibles soluciones.