Examinando por Autor "Allende, Miguel L."

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    A high-throughput chemically induced inflammation assay in zebrafish
    (BMC, 2010-12-22) d'Alençon, Claudia A.; Peña, Oscar A.; Wittmann, Christine; Gallardo, Viviana E.; Jones, Rebecca A.; Loosli, Felix; Liebel, Urban; Grabher, Clemens; Allende, Miguel L.
    Background: Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening.Results: Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction.Conclusions: This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay.See Commentary article: http://www.biomedcentral.com/1741-7007/8/148. © 2010 d'Alençon et al; licensee BioMed Central Ltd.
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    Aloe vera reduces gut inflammation induced by soybean meal in Atlantic salmon (Salmo salar)
    (Frontiers Media S.A., 2022) Fehrmann-Cartes, Karen; Vega, Matías; Vera, Frank; Enríquez, Ricardo; Feijóo, Carmen G.; Allende, Miguel L.; Hernández, Adrián J.; Romero, Alex
    Plant-based protein sources, such as soybean, are widely used in fish nutrition due to their market availability, wide distribution and acceptable nutritional quality. However, in some fish species, soybean meal-based diets cause gut inflammation, decreasing both nutrient absorption and growth rates. A suitable alternative to avoid these problems could be the application of additives with anti-inflammatory activity to the diet. In this study, an Aloe vera (Aloe barbadensis Miller, AV) extract was analyzed as a dietary additive to reduce the gut inflammation in Atlantic salmon (Salmo salar) fed with soybean meal (SBM) diet. Fish were distributed in four duplicated groups and fed 28 days with fish meal control diet (FM), AV inclusion diet (AV), FM diet supplemented with AV (FM+AV), SBM diet to induce enteritis and SBM+AV. The fish gut response to these treatments was analyzed in distal intestine by histopathological scores, tissue morphometric measurements and immune gene expression parameters. The score results in fish fed with SBM-based diet clearly showed enteritis, meanwhile fish fed with AV supplemented diet significantly reduced the intestinal SBM signs of damage. These findings were associated to reduction of goblet cells number, lamina propria thickness and sub-epithelial mucosa size, with a significant decrease on pro-inflammatory cytokine il-1β to basal levels, similar to those present in fish fed FM diets. In conclusion, the administration of AV in salmon diet showed a protective intestinal activity against the detrimental effects of SBM, opening the possibility to improve its use as a feed additive in aquafeeds. Copyright © 2022 Fehrmann-Cartes, Vega, Vera, Enríquez, Feijóo, Allende, Hernández and Romero.
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    Drosophila DAxud1 Has a Repressive Transcription Activity on Hsp70 and Other Heat Shock Genes
    (MDPI, 2023-04) Zuñiga-Hernandez, Jorge; Meneses, Claudio; Bastias, Macarena; Allende, Miguel L.; Glavic, Alvaro
    Drosophila melanogaster DAxud1 is a transcription factor that belongs to the Cysteine Serine Rich Nuclear Protein (CSRNP) family, conserved in metazoans, with a transcriptional transactivation activity. According to previous studies, this protein promotes apoptosis and Wnt signaling-mediated neural crest differentiation in vertebrates. However, no analysis has been conducted to determine what other genes it might control, especially in connection with cell survival and apoptosis. To partly answer this question, this work analyzes the role of Drosophila DAxud1 using Targeted-DamID-seq (TaDa-seq), which allows whole genome screening to determine in which regions it is most frequently found. This analysis confirmed the presence of DAxud1 in groups of pro-apoptotic and Wnt pathway genes, as previously described; furthermore, stress resistance genes that coding heat shock protein (HSP) family genes were found as hsp70, hsp67, and hsp26. The enrichment of DAxud1 also identified a DNA-binding motif (AYATACATAYATA) that is frequently found in the promoters of these genes. Surprisingly, the following analyses demonstrated that DAxud1 exerts a repressive role on these genes, which are necessary for cell survival. This is coupled with the pro-apoptotic and cell cycle arrest roles of DAxud1, in which repression of hsp70 complements the maintenance of tissue homeostasis through cell survival modulation.
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    Epigenetic control of the bone-master Runx2 gene during osteoblast-lineage commitment by the histone demethylase JARID1B/KDM5B
    (American Society for Biochemistry and Molecular Biology Inc., 2015-12) Rojas, Adriana; Aguilar, Rodrigo; Henriquez, Berta; Lian, Jane B.; Stein, Janet L.; Stein, Gary S.; Van Wijnen, Andre J.; Van Zundert, Brigitte; Allende, Miguel L.; Montecino, Martin
    Transcription factor Runx2 controls bone development and osteoblast differentiation by regulating expression of a significant number of bone-related target genes. Here, we report that transcriptional activation and repression of the Runx2 gene via its osteoblast-specific P1 promoter (encoding mRNA for the Runx2/p57 isoform) is accompanied by selective deposition and elimination of histone marks during differentiation of mesenchymal cells to the osteogenic and myoblastic lineages. These epigenetic profiles are mediated by key components of the Trithorax/COMPASS-like and Polycomb group complexes together with histone arginine methylases like PRMT5 and lysine demethylases like JARID1B/KDM5B. Importantly, knockdown of the H3K4me2/3 demethylase JARID1B, but not of the demethylases UTX and NO66, prevents repression of the Runx2 P1 promoter during myogenic differentiation of mesenchymal cells. The epigenetically forced expression of Runx2/p57 and osteocalcin, a classical bone-related target gene, under myoblastic-differentiation is accompanied by enrichment of the H3K4me3 and H3K27ac marks at the Runx2 P1 promoter region. Our results identify JARID1B as a key component of a potent epigenetic switch that controls mesenchymal cell fate into myogenic and osteogenic lineages. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
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    Genomes of the Orestias pupfish from the Andean Altiplano shed light on their evolutionary history and phylogenetic relationships within Cyprinodontiformes
    (BioMed Central Ltd, 2024-12) Morales, Pamela; Gajardo, Felipe; Valdivieso, Camilo; Valladares, Moisés A.; Di Genova, Alex; Orellana, Ariel; Gutiérrez, Rodrigo A.; González, Mauricio; Montecino, Martin; Maass, Alejandro; Méndez, Marco A.; Allende, Miguel L.
    Background: To unravel the evolutionary history of a complex group, a comprehensive reconstruction of its phylogenetic relationships is crucial. This requires meticulous taxon sampling and careful consideration of multiple characters to ensure a complete and accurate reconstruction. The phylogenetic position of the Orestias genus has been estimated partly on unavailable or incomplete information. As a consequence, it was assigned to the family Cyprindontidae, relating this Andean fish to other geographically distant genera distributed in the Mediterranean, Middle East and North and Central America. In this study, using complete genome sequencing, we aim to clarify the phylogenetic position of Orestias within the Cyprinodontiformes order. Results: We sequenced the genome of three Orestias species from the Andean Altiplano. Our analysis revealed that the small genome size in this genus (~ 0.7 Gb) was caused by a contraction in transposable element (TE) content, particularly in DNA elements and short interspersed nuclear elements (SINEs). Using predicted gene sequences, we generated a phylogenetic tree of Cyprinodontiformes using 902 orthologs extracted from all 32 available genomes as well as three outgroup species. We complemented this analysis with a phylogenetic reconstruction and time calibration considering 12 molecular markers (eight nuclear and four mitochondrial genes) and a stratified taxon sampling to consider 198 species of nearly all families and genera of this order. Overall, our results show that phylogenetic closeness is directly related to geographical distance. Importantly, we found that Orestias is not part of the Cyprinodontidae family, and that it is more closely related to the South American fish fauna, being the Fluviphylacidae the closest sister group. Conclusions: The evolutionary history of the Orestias genus is linked to the South American ichthyofauna and it should no longer be considered a member of the Cyprinodontidae family. Instead, we submit that Orestias belongs to the Orestiidae family, as suggested by Freyhof et al. (2017), and that it is the sister group of the Fluviphylacidae family, distributed in the Amazonian and Orinoco basins. These two groups likely diverged during the Late Eocene concomitant with hydrogeological changes in the South American landscape. © The Author(s) 2024.
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    Polycomb PRC2 complex mediates epigenetic silencing of a critical osteogenic master regulator in the hippocampus
    (Elsevier B.V., 2016-08) Aguilar, Rodrigo; Bustos, Fernando J.; Saez, Mauricio; Rojas, Adriana; Allende, Miguel L.; van Wijnen, Andre J.; van Zundert, Brigitte; Montecino, Martin
    During hippocampal neuron differentiation, the expression of critical inducers of non-neuronal cell lineages must be efficiently silenced. Runx2 transcription factor is the master regulator of mesenchymal cells responsible for intramembranous osteoblast differentiation and formation of the craniofacial bone tissue that surrounds and protects the central nervous system (CNS) in mammalian embryos. The molecular mechanisms that mediate silencing of the Runx2 gene and its downstream target osteogenic-related genes in neuronal cells have not been explored. Here, we assess the epigenetic mechanisms that mediate silencing of osteoblast-specific genes in CNS neurons. In particular, we address the contribution of histone epigenetic marks and histone modifiers on the silencing of the Runx2/p57 bone-related isoform in rat hippocampal tissues at embryonic to adult stages. Our results indicate enrichment of repressive chromatin histone marks and of the Polycomb PRC2 complex at the Runx2/p57 promoter region. Knockdown of PRC2 H3K27-methyltransferases Ezh2 and Ezh1, or forced expres sion of the Trithorax/COMPASS subunit Wdr5 activates Runx2/p57 mRNA expression in both immature and mature hippocampal cells. Together these results indicate that complementary epigenetic mechanisms progressively and efficiently silence critical osteoblastic genes during hippocampal neuron differentiation. © 2016 Elsevier B.V. All rights reserved.
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    Whole Genome Sequence, Variant Discovery and Annotation in Mapuche-Huilliche Native South Americans
    (Nature Publishing Group, 2019-12) Vidal, Elena A.; Moyano, Tomás C.; Bustos, Bernabé I.; Pérez-Palma, Eduardo; Moraga, Carol; Riveras, Eleodoro; Montecinos, Alejandro; Azócar, Lorena; Soto, Daniela C.; Vidal, Mabel; Genova, Alex Di; Puschel, Klaus; Nürnberg, Peter; Buch, Stephan; Hampe, Jochen; Allende, Miguel L.; Cambiazo, Verónica; González, Mauricio; Hodar, , Christian; Montecino, Martín; Muñoz-Espinoza, Claudia; Orellana, Ariel; Reyes-Jara, Angélica; Travisany, Dante; Vizoso, Paula; Moraga, Mauricio; Eyheramendy, Susana; Maass, Alejandro; Ferrari, Giancarlo V. De; Miquel, Juan Francisco; Gutiérrez, Rodrigo A.
    Whole human genome sequencing initiatives help us understand population history and the basis of genetic diseases. Current data mostly focuses on Old World populations, and the information of the genomic structure of Native Americans, especially those from the Southern Cone is scant. Here we present annotation and variant discovery from high-quality complete genome sequences of a cohort of 11 Mapuche-Huilliche individuals (HUI) from Southern Chile. We found approximately 3.1 × 10 6 single nucleotide variants (SNVs) per individual and identified 403,383 (6.9%) of novel SNVs events. Analyses of large-scale genomic events detected 680 copy number variants (CNVs) and 4,514 structural variants (SVs), including 398 and 1,910 novel events, respectively. Global ancestry composition of HUI genomes revealed that the cohort represents a sample from a marginally admixed population from the Southern Cone, whose main genetic component derives from Native American ancestors. Additionally, we found that HUI genomes contain variants in genes associated with 5 of the 6 leading causes of noncommunicable diseases in Chile, which may have an impact on the risk of prevalent diseases in Chilean and Amerindian populations. Our data represents a useful resource that can contribute to population-based studies and for the design of early diagnostics or prevention tools for Native and admixed Latin American populations. © 2019, The Author(s).