Oligodendrocyte differentiation from adult multipotent stem cells is modulated by glutamate

We used multipotent stem cells (MSCs) derived from the young rat subventricular zone (SVZ) to study the effects of glutamate in oligodendrocyte maturation. Glutamate stimulated oligodendrocyte differentiation from SVZ-derived MSCs through the activation of specific N-methyl--aspartate (NMDA) receptor subunits. The effect of glutamate and NMDA on oligodendrocyte differentiation was evident in both the number of newly generated oligodendrocytes and their morphology. In addition, the levels of NMDAR1 and NMDAR2A protein increased during differentiation, whereas NMDAR2B and NMDAR3 protein levels decreased, suggesting differential expression of NMDA receptor subunits during maturation. Microfluorimetry showed that the activation of NMDA receptors during oligodendrocyte differentiation elevated cytosolic calcium levels and promoted myelination in cocultures with neurons. Moreover, we observed that stimulation of MSCs by NMDA receptors induced the generation of reactive oxygen species (ROS), which were negatively modulated by the NADPH inhibitor apocynin, and that the levels of ROS correlated with the degree of differentiation. Taken together, these findings suggest that ROS generated by NADPH oxidase by the activation of NMDA receptors promotes the maturation of oligodendrocytes and favors myelination

The European Research Infrastructure for Heritage Science (E-RIHS): an infrastructure for an interdisciplinary scientific domain

IV International Congress Science and Technology for the Conservation of Cultural Heritage 2019Heritage is a key component of the European identity. The study and preservation of cultural and natural heritage is a global challenge for science and the European society at large. The European Research Infrastructure for Heritage Science (E-RIHS) supports research on heritage interpretation, preservation, documentation and management. State-of-the-art tools and services will be provided by cross-disciplinary groups of researchers to cross-disciplinary users and scientific communities working to advance knowledge about heritage and to devise innovative strategies for its preservation. E-RIHS connects researchers in the humanities and natural sciences and fosters a trans-disciplinary culture of exchange and cooperation. ERIHS pursues the integration of European world-class facilities to create a cohesive entity playing a connecting role in the global community of heritage science E-RIHS will provide access through four platforms: ARCHLAB, providing access to archives of technical images, analytical data and conservation documentation of prestigious European museums and conservation and research institutions; DIGILAB, to provide virtual access to digital data for heritage science, making them FAIR (Findable, Accessible, Interoperable and Reusable); FIXLAB, providing access to large and medium-scale facilities offering unique expertise in the heritage field; and MOLAB, by which mobile laboratories including advanced mobile analytical instrumentation will be available to carry out in-situ investigations. E-RIHS was included in the ESFRI 2016 Roadmap update, and is currently working towards it establishment as a distributed research infrastructure with the legal form of a European Research Infrastructure Consortium (ERIC). I will have a star design, with a Central Hub in Florence (Italy), and National Hubs in each participating country. In this conference, the vision and structure of E-RIHS, and the advancements towards the setting up of the ERIC and the National Hubs will be presented, with a special focus on the development of the Spanish National Hub, E-RIHS.e

Construyendo E-RIHS, la infraestructura europea de investigación en ciencia del patrimonio

LA CIENCIA Y EL ARTE VII: Ciencias y tecnologías aplicadas a la conservación del patrimonioE-RIHS (>European Research Infrastructure for Heritage Science> www.e-rihs.eu) es una infraestructura para el desarrollo de la investigación en patrimonio cultural y natural en aspectos que incluyen su interpretación, conservación, documentación y gestión. E-RIHS se configura como una infraestructura distribuida, con una Sede Central (previsiblemente en Florencia, Italia) y Nodos Nacionales. Proveerá de herramientas y servicios avanzados >desarrollados por grupos de investigación trans-disciplinares> a usuarios científicos que trabajen en el avance del conocimiento sobre el patrimonio y en estrategias innovadoras para su preservación. A través de procedimientos concertados entre los participantes, E-RIHS proporcionará un acceso integrado a las tecnologías de análisis más punteras y a archivos científicos a través de cuatro plataformas: FIXLAB, MOLAB, ARCHLAB y DIGILAB. En este trabajo se presenta E-RIHS, los avances en establecimiento de la infraestructura europea y en la creación y organización del Nodo Nacional español, E-RIHS.e

Characterisation of a flavonoid ligand of the fungal protein Alt a 1

Spores of pathogenic fungi are virtually ubiquitous and cause human disease and severe losses in crops. The endophytic fungi Alternaria species produce host-selective phytotoxins. Alt a 1 is a strongly allergenic protein found in A. alternata that causes severe asthma. Despite the well-established pathogenicity of Alt a 1, the molecular mechanisms underlying its action and physiological function remain largely unknown. To gain insight into the role played by this protein in the pathogenicity of the fungus, we studied production of Alt a 1 and its activity in spores. We found that Alt a 1 accumulates inside spores and that its release with a ligand is pH-dependent, with optimum production in the 5.0-6.5 interval. The Alt a 1 ligand was identified as a methylated flavonoid that inhibits plant root growth and detoxifies reactive oxygen species. We also found that Alt a 1 changes its oligomerization state depending on the pH of the surrounding medium and that these changes facilitate the release of the ligand. Based on these results, we propose that release of Alt a 1 should be a pathogenic target in approaches used to block plant defenses and consequently to favor fungal entry into the plant

Ependymal cells along the lateral ventricle express functional P2X7 receptors

Ependymal cells line the cerebral ventricles and are located in an ideal position to detect central nervous system injury and inflammation. The signaling mechanisms of ependymal cells, however, are poorly understood. As extracellular adenosine 5′-triphosphate is elevated in the context of cellular damage, experiments were conducted to determine whether ependymal cells along the mouse subventricular zone (SVZ) express functional purinergic receptors. Using whole-cell patch clamp recording, widespread expression of P2X7 receptors was detected on ependymal cells based on their antagonist sensitivity profile and absence of response in P2X7−/− mice. Immunocytochemistry confirmed the expression of P2X7 receptors, and electron microscopy demonstrated that P2X7 receptors are expressed on both cilia and microvilli. Ca2+ imaging showed that P2X7 receptors expressed on cilia are indeed functional. As ependymal cells are believed to function as partner cells in the SVZ neurogenic niche, P2X7 receptors may play a role in neural progenitor response to injury and inflammation

FACS isolation of endothelial cells and pericytes from mouse brain microregions

The vasculature is emerging as a key contributor to brain function during neurodevelopment and in mature physiological and pathological states. The brain vasculature itself also exhibits regional heterogeneity, highlighting the need to develop approaches for purifying cells from different microregions. Previous approaches for isolation of endothelial cells and pericytes have predominantly required transgenic mice and large amounts of tissue, and have resulted in impure populations. In addition, the prospective purification of brain pericytes has been complicated by the fact that widely used pericyte markers are also expressed by other cell types in the brain. Here, we describe the detailed procedures for simultaneous isolation of pure populations of endothelial cells and pericytes directly from adult mouse brain microregions using fluorescence-activated cell sorting (FACS) with antibodies against CD31 (endothelial cells) and CD13 (pericytes). This protocol is scalable and takes ∼5 h, including microdissection of the region of interest, enzymatic tissue dissociation, immunostaining, and FACS. This protocol allows the isolation of brain vascular cells from any mouse strain under diverse conditions; these cells can be used for multiple downstream applications, including in vitro and in vivo experiments, and transcriptomic, proteomic, metabolomic, epigenomic, and single-cell analysis