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A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle

Muscular manifestation of myotonic dystrophy type 1 (DM1), a common inheritable degenerative multisystem disorder, is mainly caused by expression of RNA from a (CTG·CAG)(n)-expanded DM1 locus. Here, we report on comparative profiling of expression of normal and expanded endogenous or transgenic tran...

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Autores principales: Gudde, Anke E. E. G., González-Barriga, Anchel, van den Broek, Walther J. A. A., Wieringa, Bé, Wansink, Derick G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4805313/
https://www.ncbi.nlm.nih.gov/pubmed/26908607
http://dx.doi.org/10.1093/hmg/ddw042
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author Gudde, Anke E. E. G.
González-Barriga, Anchel
van den Broek, Walther J. A. A.
Wieringa, Bé
Wansink, Derick G.
author_facet Gudde, Anke E. E. G.
González-Barriga, Anchel
van den Broek, Walther J. A. A.
Wieringa, Bé
Wansink, Derick G.
author_sort Gudde, Anke E. E. G.
collection PubMed
description Muscular manifestation of myotonic dystrophy type 1 (DM1), a common inheritable degenerative multisystem disorder, is mainly caused by expression of RNA from a (CTG·CAG)(n)-expanded DM1 locus. Here, we report on comparative profiling of expression of normal and expanded endogenous or transgenic transcripts in skeletal muscle cells and biopsies from DM1 mouse models and patients in order to help us in understanding the role of this RNA-mediated toxicity. In tissue of HSA(LR) mice, the most intensely used ‘muscle-only’ model in the DM1 field, RNA from the α-actin (CTG)250 transgene was at least 1000-fold more abundant than that from the Dmpk gene, or the DMPK gene in humans. Conversely, the DMPK transgene in another line, DM500/DMSXL mice, was expressed ∼10-fold lower than the endogenous gene. Temporal regulation of expanded RNA expression differed between models. Onset of expression occurred remarkably late in HSA(LR) myoblasts during in vitro myogenesis whereas Dmpk or DMPK (trans)genes were expressed throughout proliferation and differentiation phases. Importantly, quantification of absolute transcript numbers revealed that normal and expanded Dmpk/DMPK transcripts in mouse models and DM1 patients are low-abundance RNA species. Northern blotting, reverse transcriptase–quantitative polymerase chain reaction, RNA-sequencing and fluorescent in situ hybridization analyses showed that they occur at an absolute number between one and a few dozen molecules per cell. Our findings refine the current RNA dominance theory for DM1 pathophysiology, as anomalous factor binding to expanded transcripts and formation of soluble or insoluble ribonucleoprotein aggregates must be nucleated by only few expanded DMPK transcripts and therefore be a small numbers game.
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spelling pubmed-48053132016-03-24 A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle Gudde, Anke E. E. G. González-Barriga, Anchel van den Broek, Walther J. A. A. Wieringa, Bé Wansink, Derick G. Hum Mol Genet Articles Muscular manifestation of myotonic dystrophy type 1 (DM1), a common inheritable degenerative multisystem disorder, is mainly caused by expression of RNA from a (CTG·CAG)(n)-expanded DM1 locus. Here, we report on comparative profiling of expression of normal and expanded endogenous or transgenic transcripts in skeletal muscle cells and biopsies from DM1 mouse models and patients in order to help us in understanding the role of this RNA-mediated toxicity. In tissue of HSA(LR) mice, the most intensely used ‘muscle-only’ model in the DM1 field, RNA from the α-actin (CTG)250 transgene was at least 1000-fold more abundant than that from the Dmpk gene, or the DMPK gene in humans. Conversely, the DMPK transgene in another line, DM500/DMSXL mice, was expressed ∼10-fold lower than the endogenous gene. Temporal regulation of expanded RNA expression differed between models. Onset of expression occurred remarkably late in HSA(LR) myoblasts during in vitro myogenesis whereas Dmpk or DMPK (trans)genes were expressed throughout proliferation and differentiation phases. Importantly, quantification of absolute transcript numbers revealed that normal and expanded Dmpk/DMPK transcripts in mouse models and DM1 patients are low-abundance RNA species. Northern blotting, reverse transcriptase–quantitative polymerase chain reaction, RNA-sequencing and fluorescent in situ hybridization analyses showed that they occur at an absolute number between one and a few dozen molecules per cell. Our findings refine the current RNA dominance theory for DM1 pathophysiology, as anomalous factor binding to expanded transcripts and formation of soluble or insoluble ribonucleoprotein aggregates must be nucleated by only few expanded DMPK transcripts and therefore be a small numbers game. Oxford University Press 2016-04-15 2016-02-16 /pmc/articles/PMC4805313/ /pubmed/26908607 http://dx.doi.org/10.1093/hmg/ddw042 Text en © The Author 2016. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Articles
Gudde, Anke E. E. G.
González-Barriga, Anchel
van den Broek, Walther J. A. A.
Wieringa, Bé
Wansink, Derick G.
A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title_full A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title_fullStr A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title_full_unstemmed A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title_short A low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
title_sort low absolute number of expanded transcripts is involved in myotonic dystrophy type 1 manifestation in muscle
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4805313/
https://www.ncbi.nlm.nih.gov/pubmed/26908607
http://dx.doi.org/10.1093/hmg/ddw042
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