23 research outputs found

    El Jardín Botánico Mediterráneo de la ETSI Agrónomos: Caracterización de la flora herbácea en relación a la flora arbóreo-arbustiva.

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    En este trabajo se ha caracterizado la flora herbácea del Jardín Botánico Mediterráneo situado en los Campos de Prácticas de la ETSI Agrónomos de Madrid. Se ha realizado tanto para el Jardín en su conjunto como para cada una de las formaciones vegetales representadas y de forma comparativa entre ellas. Considerando el Jardín en su conjunto o determinada comunidad vegetal, los resultados muestran un predominio de tres-cuatro familias botánicas. Predominan los biotipos terofítico y hemicriptofítico, la indiferencia edáfica y la clase fitosociológica Ruderali-Secalietea. Se ha podido corroborar que la flora herbácea del Jardín es principalmente de carácter arvense-ruderal, nitrófila e invasora

    The NAD+-Dependent SIRT1 Deacetylase Translates a Metabolic Switch into Regulatory Epigenetics in Skeletal Muscle Stem Cells

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    SummaryStem cells undergo a shift in metabolic substrate utilization during specification and/or differentiation, a process that has been termed metabolic reprogramming. Here, we report that during the transition from quiescence to proliferation, skeletal muscle stem cells experience a metabolic switch from fatty acid oxidation to glycolysis. This reprogramming of cellular metabolism decreases intracellular NAD+ levels and the activity of the histone deacetylase SIRT1, leading to elevated H4K16 acetylation and activation of muscle gene transcription. Selective genetic ablation of the SIRT1 deacetylase domain in skeletal muscle results in increased H4K16 acetylation and deregulated activation of the myogenic program in SCs. Moreover, mice with muscle-specific inactivation of the SIRT1 deacetylase domain display reduced myofiber size, impaired muscle regeneration, and derepression of muscle developmental genes. Overall, these findings reveal how metabolic cues can be mechanistically translated into epigenetic modifications that regulate skeletal muscle stem cell biology

    FoxO maintains a genuine muscle stem-cell quiescent state until geriatric age

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    Tissue regeneration declines with ageing but little is known about whether this arises from changes in stem-cell heterogeneity. Here, in homeostatic skeletal muscle, we identify two quiescent stem-cell states distinguished by relative CD34 expression: CD34High, with stemness properties (genuine state), and CD34Low, committed to myogenic differentiation (primed state). The genuine-quiescent state is unexpectedly preserved into later life, succumbing only in extreme old age due to the acquisition of primed-state traits. Niche-derived IGF1-dependent Akt activation debilitates the genuine stem-cell state by imposing primed-state features via FoxO inhibition. Interventions to neutralize Akt and promote FoxO activity drive a primed-to-genuine state conversion, whereas FoxO inactivation deteriorates the genuine state at a young age, causing regenerative failure of muscle, as occurs in geriatric mice. These findings reveal transcriptional determinants of stem-cell heterogeneity that resist ageing more than previously anticipated and are only lost in extreme old age, with implications for the repair of geriatric muscle.The authors acknowledge funding from MINECO-Spain (grant no. RTI2018-096068), ERC2016-AdG-741966, LaCaixa-HEALTH-HR17-00040, MDA, UPGRADE-H2020-825825, AFM and DPP-Spain to P.M.-C; María-de-Maeztu-Program for Units of Excellence to UPF (grant no. MDM-2014-0370) and the Severo-Ochoa-Program for Centers of Excellence to CNIC (grant no. SEV-2015-0505). This work was also supported by NIAMS IRP through NIH grants nos AR041126 and AR041164 to V.S. and utilized computational resources of the NIH HPC Biowulf cluster (http://hpc.nih.gov); ASI, Ricerca Finalizzata, Ateneo Sapienza to A.M.; AIRC (grant no. 23257); ASI (grant no. MARS-PRE, DC-VUM-2017-006); H2020-MSCA-RISE-2014 (645648) to M.S. and a FNR core grant (grant no. C15/BM/10397420) to A.d.S. L.G.P. was partially supported by an FPI fellowship and an EMBO fellowship (grant no. ALTF 420-2017); and S.C., X.H. and V.M. by FI, Severo-Ochoa and PFI Fellowships (Spain), respectively

    Mir-214-Dependent Regulation of the Polycomb Protein Ezh2 in Skeletal Muscle and Embryonic Stem Cells

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    Arthur Manuscript date: 2010 October 9Polycomb group (PcG) proteins exert essential functions in the most disparate biological processes. The contribution of PcG proteins to cell commitment and differentiation relates to their ability to repress transcription of developmental regulators in embryonic stem (ES) cells and in committed cell lineages, including skeletal muscle cells (SMC). PcG proteins are preferentially removed from transcribed regions, but the underlying mechanisms remain unclear. Here, PcG proteins are found to occupy and repress transcription from an intronic region containing the microRNA miR-214 in undifferentiated SMC. Differentiation coincides with PcG disengagement, recruitment of the developmental regulators MyoD and myogenin, and activation of miR-214 transcription. Once transcribed, miR-214 negatively feeds back on PcG by targeting the Ezh2 3′UTR, the catalytic subunit of the PRC2 complex. miR-214-mediated Ezh2 protein reduction accelerates SMC differentiation and promotes unscheduled transcription of developmental regulators in ES cells. Thus, miR-214 and Ezh2 establish a regulatory loop controlling PcG-dependent gene expression during differentiation.National Institute of Arthritis and Musculoskeletal and Skin Diseases (U.S.) (Intramural Research Program

    Protocol for RNA-seq library preparation starting from a rare muscle stem cell population or a limited number of mouse embryonic stem cells

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    It remains challenging to generate reproducible, high-quality cDNA libraries from RNA derived from rare cell populations. Here, we describe a protocol for high-throughput RNA-seq library preparation, including isolation of 200 skeletal muscle stem cells from mouse tibialis anterior muscle by fluorescence-activated cell sorting and cDNA preparation. We also describe RNA extraction and cDNA preparation from differentiating mouse embryonic stem cells. For complete details on the use and execution of this protocol, please refer to Juan et al. (2016) and Garcia-Prat et al. (2016).Funds to V.S. were provided by the NIAMS-IRP through NIH grants AR041126 and AR041164. Work in P.M.-C’s laboratory was supported by MINECO-RTI2018-096068, ERC-AdG-741966, LaCaixa-HEALTH-HR17-00040, UPGRADE-H2020-825825 and Marató-TV3, MDM-2014-0370, and SEV-2015-0505. Authors acknowledge technical support from the UPF/CRG and CNIC flow cytometry units

    Rapid deglaciation of the La Vega gorge (Sierra de Gredos, Iberian Peninsula) at the end of the global Last Glacial Maximum

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    International audienceAbstract Previous studies from the Iberian Central System and other mountains of the Iberian Peninsula and Europe suggest that deglaciation in this area occurred at the end or immediately after the global Last Glacial Maximum (LGM, 26.5–19 ka). In this research, we investigate the timing and speed of deglaciation of the palaeoglacier in La Vega gorge (Iberian Central system) since the global LGM, dating the outer moraines, glacially polished bedrock and glaciofluvial deposits by means of in situ cosmogenic 10 Be and optically stimulated luminescence. The results show that one intermediate arc located at the outer frontal moraine system has an age of ~21 ka, being consistent with the global LGM. Deglaciation began at ~21–19 ka, speeding up at ~19 ka. We estimate that around 4–5 km of the palaeoglacier receded in ~1–2 kyrs since ~19 ka, leaving La Vega gorge probably ice‐free at ~18 ka. Therefore, our data confirm that deglaciation in Sierra de Gredos began at the end of the global LGM, characterised by a rapid and massive retreat of glaciers

    Polycomb EZH2 controls self-renewal and safeguards the transcriptional identity of skeletal muscle stem cells

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    Satellite cells (SCs) sustain muscle growth and empower adult skeletal muscle with vigorous regenerative abilities. Here, we report that EZH2, the enzymatic subunit of the Polycomb-repressive complex 2 (PRC2), is expressed in both Pax7+/Myf5− stem cells and Pax7+/Myf5+ committed myogenic precursors and is required for homeostasis of the adult SC pool. Mice with conditional ablation of Ezh2 in SCs have fewer muscle postnatal Pax7+ cells and reduced muscle mass and fail to appropriately regenerate. These defects are associated with impaired SC proliferation and derepression of genes expressed in nonmuscle cell lineages. Thus, EZH2 controls self-renewal and proliferation, and maintains an appropriate transcriptional program in SCs

    Roles of H3K27me2 and H3K27me3 Examined during Fate Specification of Embryonic Stem Cells

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    Summary: The polycomb repressive complex 2 (PRC2) methylates lysine 27 of histone H3 (H3K27) through its catalytic subunit Ezh2. PRC2-mediated di- and tri-methylation (H3K27me2/H3K27me3) have been interchangeably associated with gene repression. However, it remains unclear whether these two degrees of H3K27 methylation have different functions. In this study, we have generated isogenic mouse embryonic stem cells (ESCs) with a modified H3K27me2/H3K27me3 ratio. Our findings document dynamic developmental control in the genomic distribution of H3K27me2 and H3K27me3 at regulatory regions in ESCs. They also reveal that modifying the ratio of H3K27me2 and H3K27me3 is sufficient for the acquisition and repression of defined cell lineage transcriptional programs and phenotypes and influences induction of the ESC ground state. : Juan et al. use genome editing to subtly modify H3K27 methylation states (H3K27me2/H3K27me3) and report that such perturbation influences the ability of ESCs to differentiate into preferential cell lineages and acquire a “ground state.” Keywords: polycomb proteins, H3K27 methylation, embryonic stem cell
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