Alterations in the muscle force transfer apparatus in aged rats during unloading and reloading: impact of microRNA‐31

Hughes, David C.; Marcotte, George R.; Baehr, Leslie M.; West, Daniel W. D.; Marshall, Andrea G.; Ebert, Scott M.; Davidyan, Arik; Adams, Christopher M.; Bodine, Sue C.; Baar, Keith

doi:10.1113/jp275833

Cited by 23 publications

(25 citation statements)

References 61 publications

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“…For these experiments as a result of reference gene instability, after first assessing total RNA per mg tissue and then taking the C t expression value, the mRNA was described relative to tissue weight and subsequently reported as the fold change relative to control muscles, as described previously (Heinemeier et al 2009). An analysis that has previously been used extensively by the group (Baehr et al 2016;Baehr et al 2017;Hughes et al 2018).…”

Section: Strength Measurementmentioning

confidence: 99%

“…740424.50; Macherey-Nagel, Düren, Germany) in accordance with the manufacturer's instructions. The electroporation of DNA plasmids was then performed as described previously (Ebert et al 2010;Hughes et al 2018). Briefly, after 2 h of pretreatment with 0.4 U µL -1 of hyaluronidase, 20 µg of mIR RNAi or negative control plasmid was injected into the TA muscle in C57/BL6 male mice (12-16 weeks old, n = 5; purchased from Charles River Laboratories) and the hind limbs were placed between two-paddle electrodes and subjected to 10 pulses (20 ms) of 175 V cm -1 using an ECM-830 electroporator (BTX Harvard Apparatus, Holliston, MA, USA).…”

Section: Muscle Specific Knockdown Of Ubr5 For Antibody Validationmentioning

confidence: 99%

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UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

Seaborne

Hughes

Turner

et al. 2019

The Journal of Physiology

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Key points We have recently identified that a HECT domain E3 ubiquitin ligase, named UBR5, is altered epigenetically (via DNA methylation) after human skeletal muscle hypertrophy, where its gene expression is positively correlated with increasing lean leg mass after training and retraining. In the present study we extensively investigate this novel and uncharacterised E3 ubiquitin ligase (UBR5) in skeletal muscle atrophy, recovery from atrophy and injury, anabolism and hypertrophy. We demonstrated that UBR5 was epigenetically altered via DNA methylation during recovery from atrophy. We also determined that UBR5 was alternatively regulated versus well characterised E3 ligases, MuRF1/MAFbx, at the gene expression level during atrophy, recovery from atrophy and hypertrophy. UBR5 also increased at the protein level during recovery from atrophy and injury, hypertrophy and during human muscle cell differentiation. Finally, in humans, genetic variations of the UBR5 gene were strongly associated with larger fast‐twitch muscle fibres and strength/power performance versus endurance/untrained phenotypes. Abstract We aimed to investigate a novel and uncharacterized E3 ubiquitin ligase in skeletal muscle atrophy, recovery from atrophy/injury, anabolism and hypertrophy. We demonstrated an alternate gene expression profile for UBR5 vs. well characterized E3‐ligases, MuRF1/MAFbx, where, after atrophy evoked by continuous‐low‐frequency electrical‐stimulation in rats, MuRF1/MAFbx were both elevated, yet UBR5 was unchanged. Furthermore, after recovery of muscle mass post TTX‐induced atrophy in rats, UBR5 was hypomethylated and increased at the gene expression level, whereas a suppression of MuRF1/MAFbx was observed. At the protein level, we also demonstrated a significant increase in UBR5 after recovery of muscle mass from hindlimb unloading in both adult and aged rats, as well as after recovery from atrophy evoked by nerve crush injury in mice. During anabolism and hypertrophy, UBR5 gene expression increased following acute loading in three‐dimensional bioengineered mouse muscle in vitro, and after chronic electrical stimulation‐induced hypertrophy in rats in vivo, without increases in MuRF1/MAFbx. Additionally, UBR5 protein abundance increased following functional overload‐induced hypertrophy of the plantaris muscle in mice and during differentiation of primary human muscle cells. Finally, in humans, genetic association studies (>700,000 single nucleotide polymorphisms) demonstrated that the A alleles of rs10505025 and rs4734621 single nucleotide polymorphisms in the UBR5 gene were strongly associated with larger cross‐sectional area of fast‐twitch muscle fibres and favoured strength/power vs. endurance/untrained phenotypes. Overall, we suggest that: (i) UBR5 comprises a novel E3 ubiquitin ligase that is inversely regulated to MuRF1/MAFbx; (ii) UBR5 is epigenetically regulated; and (iii) UBR5 is elevated at both the gene expression and protein level during recovery from skeletal muscle atrophy and hypertrophy.

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Section: Strength Measurementmentioning

confidence: 99%

Section: Muscle Specific Knockdown Of Ubr5 For Antibody Validationmentioning

confidence: 99%

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

Seaborne

Hughes

Turner

et al. 2019

The Journal of Physiology

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“…We also observed significant reductions in dystrophin protein content in Fbxl22 transfected muscles. Dystrophin is essential for lateral force transmission and protecting skeletal muscle from contraction-induced injury (29,(39)(40)(41). The alterations in these sarcomeric proteins coincided with the development of a degenerative muscle fiber phenotype in Fbxl22 transfected tissues.…”

Section: Discussionmentioning

confidence: 98%

“…As a result of reference gene instability, gene expression was normalized to tissue weight and subsequently reported as the fold change relative to control muscles, as described previously (67). This type of analysis has previously been used extensively by our group (40,52,53).…”

Section: Gene Expression By Quantitative Rt-pcr In Skeletal Musclementioning

confidence: 99%

Identification and Characterization of Fbxl22, a novel skeletal muscle atrophy-promoting E3 ubiquitin ligase

Hughes

Baehr

Driscoll

et al. 2020

Preprint

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Muscle-specific E3 ubiquitin ligases have been identified in muscle atrophy-inducing conditions. The purpose of the current study was to explore the functional role of Fbxl22, and a newly identified splice variant , in skeletal muscle homeostasis and neurogenic muscle atrophy. The full-length gene and novel splice variant are transcriptionally induced early (after 3 days) during neurogenic muscle atrophy. In vivo overexpression of Fbxl22 isoforms in mouse skeletal muscle lead to evidence of myopathy/atrophy suggesting that both are involved in the process of neurogenic muscle atrophy. Knockdown of Fbxl22 in MuRF1 KO muscles resulted in significant additive muscle sparing at 7 days of denervation. Targeting two E3 ubiquitin ligases appears to have a strong additive effect on protecting muscle mass loss with denervation and these findings have important implications in the development of therapeutic strategies to treat muscle atrophy.

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“…Primer sequences for genes of interest and reference genes across mouse, rat and human samples are included in As above, melt curve analysis at the end of the PCR cycling protocol yielded a single peak. For these experiments due to reference gene instability, after first assessing total RNA/mg tissue and then taking the ∆ Ct expression value, the mRNA was described relative to tissue weight, and then described as fold change relative to control muscles as previously described (Heinemeier et al, 2009) an analysis used extensively by the group previously (Baehr et al, 2016;Baehr et al, 2017;Hughes et al, 2018).…”

Section: Strength Measurementmentioning

confidence: 99%

UBR5 is a Novel E3 Ubiquitin Ligase involved in Skeletal Muscle Hypertrophy and Recovery from Atrophy

Seaborne

Turner

et al. 2019

Preprint

View full text Add to dashboard Cite

We aimed to investigate a novel and uncharacterised E3 ubiquitin ligase in skeletal muscle atrophy, recovery from atrophy/injury, anabolism and hypertrophy. We demonstrated an alternate gene expression profile for UBR5 versus well characterised E3-ligases, MuRF1/MAFbx, where after atrophy evoked by continuous-low-frequency electrical-stimulation in rats, MuRF1/MAFbx were both elevated yet UBR5 was unchanged. Furthermore, after recovery of muscle mass post tetrodotoxin (TTX) induced-atrophy in rats, UBR5 was hypomethylated and increased at the gene expression level, while a suppression of MuRF1/MAFbx was observed. At the protein level, we also demonstrated a significant increase in UBR5 after recovery of muscle mass from hindlimb unloading in both adult and aged rats, and after recovery from atrophy evoked by nerve crush injury in mice. During anabolism and hypertrophy, UBR5 gene expression increased following acute loading in three-dimensional bioengineered mouse muscle in-vitro, and after chronic electrical-stimulation-induced hypertrophy in rats in-vivo, without increases in MuRF1/MAFbx. Additionally, UBR5 protein abundance increased following functional overload-induced hypertrophy of the plantaris muscle in mice and during differentiation of primary human muscle cells. Finally, in humans, genetic association studies (>700,000 SNPs) demonstrated that the A alleles of rs10505025 and rs4734621 SNPs in the UBR5 gene were strongly associated with larger cross-sectional area of fast-twitch muscle fibres and favoured strength/power versus endurance/untrained phenotypes. Overall, we suggest that UBR5 is a novel E3 ubiquitin ligase that is inversely regulated to MuRF1/MAFbx, is epigenetically regulated, and is elevated at both the gene expression and protein level during recovery from skeletal muscle atrophy and hypertrophy.

show abstract

Alterations in the muscle force transfer apparatus in aged rats during unloading and reloading: impact of microRNA‐31

Cited by 23 publications

References 61 publications

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

Identification and Characterization of Fbxl22, a novel skeletal muscle atrophy-promoting E3 ubiquitin ligase

UBR5 is a Novel E3 Ubiquitin Ligase involved in Skeletal Muscle Hypertrophy and Recovery from Atrophy

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