Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization
In this study we surveyed a rat skeletal muscle RNA-Seq for genes that are induced by hindlimb immobilization and, in turn, become attenuated by leucine supplementation. This approach, in search of leucine-atrophy protection mediating genes, identified histone deacetylase 4 (<i>HDAC4</i>...
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2020-12-01
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author | Paula K. N. Alves André Cruz William J. Silva Siegfried Labeit Anselmo S. Moriscot |
author_facet | Paula K. N. Alves André Cruz William J. Silva Siegfried Labeit Anselmo S. Moriscot |
author_sort | Paula K. N. Alves |
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description | In this study we surveyed a rat skeletal muscle RNA-Seq for genes that are induced by hindlimb immobilization and, in turn, become attenuated by leucine supplementation. This approach, in search of leucine-atrophy protection mediating genes, identified histone deacetylase 4 (<i>HDAC4</i>) as highly responsive to both hindlimb immobilization and leucine supplementation. We then examined the impact of leucine on <i>HDAC4</i> expression, tissue localization, and target genes. A total of 76 male Wistar rats (~280 g) were submitted to hindlimb immobilization and/or leucine supplementation for 3, 7 and 12 days. These animals were euthanized, and soleus muscle was removed for further analysis. RNA-Seq analysis of hindlimb immobilized rats indicated a sharp induction (log2 = 3.4) of <i>HDAC4</i> expression which was attenuated by leucine supplementation (~50%). Real-time PCR and protein expression analysis by Western blot confirmed increased <i>HDAC4</i> mRNA after 7 days of hindlimb immobilization and mitigation of induction by leucine supplementation. Regarding the <i>HDAC4</i> localization, the proportion of positive nuclei was higher in the immobilized group and decreased after leucine supplementation. Also, we found a marked decrease of myogenin and <i>MAFbx</i>-<i>atrogin-1</i> mRNA levels upon leucine supplementation, while <i>CAMKII</i> and <i>DACH2</i> mRNA levels were increased by leucine supplementation. Our data suggest that <i>HDAC4</i> inhibition might be involved in the anti-atrophic effects of leucine. |
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spelling | doaj.art-b882c8203cf9491da5868f42034001952023-11-20T23:14:28ZengMDPI AGCells2073-44092020-12-01912258210.3390/cells9122582Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb ImmobilizationPaula K. N. Alves0André Cruz1William J. Silva2Siegfried Labeit3Anselmo S. Moriscot4Department of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo 05508000, BrazilDepartment of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo 05508000, BrazilDepartment of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo 05508000, BrazilFaculty for Clinical Medicine Mannheim of the University of Heidelberg, Institute for Integrative Pathophysiology, Universitätsmedizin Mannheim, 68169 Mannheim, GermanyDepartment of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo 05508000, BrazilIn this study we surveyed a rat skeletal muscle RNA-Seq for genes that are induced by hindlimb immobilization and, in turn, become attenuated by leucine supplementation. This approach, in search of leucine-atrophy protection mediating genes, identified histone deacetylase 4 (<i>HDAC4</i>) as highly responsive to both hindlimb immobilization and leucine supplementation. We then examined the impact of leucine on <i>HDAC4</i> expression, tissue localization, and target genes. A total of 76 male Wistar rats (~280 g) were submitted to hindlimb immobilization and/or leucine supplementation for 3, 7 and 12 days. These animals were euthanized, and soleus muscle was removed for further analysis. RNA-Seq analysis of hindlimb immobilized rats indicated a sharp induction (log2 = 3.4) of <i>HDAC4</i> expression which was attenuated by leucine supplementation (~50%). Real-time PCR and protein expression analysis by Western blot confirmed increased <i>HDAC4</i> mRNA after 7 days of hindlimb immobilization and mitigation of induction by leucine supplementation. Regarding the <i>HDAC4</i> localization, the proportion of positive nuclei was higher in the immobilized group and decreased after leucine supplementation. Also, we found a marked decrease of myogenin and <i>MAFbx</i>-<i>atrogin-1</i> mRNA levels upon leucine supplementation, while <i>CAMKII</i> and <i>DACH2</i> mRNA levels were increased by leucine supplementation. Our data suggest that <i>HDAC4</i> inhibition might be involved in the anti-atrophic effects of leucine.https://www.mdpi.com/2073-4409/9/12/2582atrophyskeletal muscle<i>HDAC4</i><i>MAFbx</i><i>MYOG</i>leucine supplementation |
spellingShingle | Paula K. N. Alves André Cruz William J. Silva Siegfried Labeit Anselmo S. Moriscot Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization Cells atrophy skeletal muscle <i>HDAC4</i> <i>MAFbx</i> <i>MYOG</i> leucine supplementation |
title | Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization |
title_full | Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization |
title_fullStr | Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization |
title_full_unstemmed | Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization |
title_short | Leucine Supplementation Decreases <i>HDAC4</i> Expression and Nuclear Localization in Skeletal Muscle Fiber of Rats Submitted to Hindlimb Immobilization |
title_sort | leucine supplementation decreases i hdac4 i expression and nuclear localization in skeletal muscle fiber of rats submitted to hindlimb immobilization |
topic | atrophy skeletal muscle <i>HDAC4</i> <i>MAFbx</i> <i>MYOG</i> leucine supplementation |
url | https://www.mdpi.com/2073-4409/9/12/2582 |
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