7 research outputs found
Expanded CTG repeats trigger miRNA alterations in Drosophila that are conserved in myotonic dystrophy type 1 patients
Myotonic dystrophy type 1 (DM1) is caused by the expansion of CTG repeats in the 3' untranslated region of the DMPK gene. Several missplicing events and transcriptional alterations have been described in DM1 patients. A large number of these defects have been reproduced in animal models expressing CTG repeats alone. Recent studies have also reported miRNA dysregulation in DM1 patients. In this work, a Drosophila model was used to investigate miRNA transcriptome alterations in the muscle, specifically triggered by CTG expansions. Twenty miRNAs were differentially expressed in CTG-expressing flies. Of these, 19 were down-regulated, whereas 1 was up-regulated. This trend was confirmed for those miRNAs conserved between Drosophila and humans (miR-1, miR-7 and miR-10) in muscle biopsies from DM1 patients. Consistently, at least seven target transcripts of these miRNAs were up-regulated in DM1 skeletal muscles. The mechanisms involved in dysregulation of miR-7 included a reduction of its primary precursor both in CTG-expressing flies and in DM1 patients. Additionally, a regulatory role for Muscleblind (Mbl) was also suggested for miR-1 and miR-7, as these miRNAs were down-regulated in flies where Mbl had been silenced. Finally, the physiological relevance of miRNA dysregulation was demonstrated for miR-10, since over-expression of this miRNA in Drosophila extended the lifespan of CTG-expressing flies. Taken together, our results contribute to our understanding of the origin and the role of miRNA alterations in DM1. © The Author 2012. Published by Oxford University Press. All rights reserved.Fundacion Ramon Areces; Generalitat Valenciana (Prometeo/2010/081); Ministerio de Ciencia e Innovacion (SAF2006-09121) in collaboration with the biotechnology company Sistemas Genomicos S.L.; FIS (FIS09-00660) ; Isabel Gemio Foundation; Accion Especial de Enfermedades Raras âCetegenâ by Genoma Espana Foundation; Generalitat Valenciana; Fundacion Ramon Areces; Banca Civica; Basque Government (AE-BFI-08.164); ISCIII; Ministerio de Economia y CompetitividadPeer Reviewe
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A quantitative gibberellin signaling biosensor reveals a role for gibberellins in internode specification at the shoot apical meristem.
Acknowledgements: We thank the colleagues of the Laboratoire Reproduction et Développement des Plantes and the Institut de Biologie Moléculaire des Plantes for fruitful discussions on this work and for sharing material and advice. We thank Mathilde Sirlin-Josserand for her help with graphical representation and statistics in Figs. 1f, g, and 2c, and in Supplementary Fig. 2g. We also thank Miguel Perez-Amador, Jan Lohmann, Yvon Jaillais, Tai-ping Sun, Miguel Blazquez, David Alabadi, and the NASC for seeds and plasmids. We thank the personnel of SFR Biosciences (UMS3444/CNRS, US8/Inserm, ENS de Lyon, UCBL) facility PLATIM, and especially Jacques Brocard, for assistance with microscopy and image analysis. We also thank Claire Lionnet for her help with image analysis. This work was supported by the ANR-16-CE13-0014 (GrowthDynamics) grant to T.V. and P.A.; a European Research Council grant (GAtransport) to R.W.; a Guangdong Laboratory for Lingnan Modern Agriculture Grant NG2021001 to B.S.; a grant from the Israel Science Foundation (grant no. 1057/21) to R.W.Funder: Guangdong Laboratory for Lingnan Modern Agriculture Grant NG2021001Growth at the shoot apical meristem (SAM) is essential for shoot architecture construction. The phytohormones gibberellins (GA) play a pivotal role in coordinating plant growth, but their role in the SAM remains mostly unknown. Here, we developed a ratiometric GA signaling biosensor by engineering one of the DELLA proteins, to suppress its master regulatory function in GA transcriptional responses while preserving its degradation upon GA sensing. We demonstrate that this degradation-based biosensor accurately reports on cellular changes in GA levels and perception during development. We used this biosensor to map GA signaling activity in the SAM. We show that high GA signaling is found primarily in cells located between organ primordia that are the precursors of internodes. By gain- and loss-of-function approaches, we further demonstrate that GAs regulate cell division plane orientation to establish the typical cellular organization of internodes, thus contributing to internode specification in the SAM
Expanded CTG repeats trigger miRNA alterations in Drosophila that are conserved in myotonic dystrophy type 1 patients
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion