Mouse soleus (slow) muscle shows greater intramyocellular lipid droplet accumulation than EDL (fast) muscle: fiber type-specific analysis

Komiya, Yusuke; Sawano, Shoko; Mashima, Daisuke; Ichitsubo, Riho; Nakamura, Mako; Tatsumi, Ryuichi; Ikeuchi, Yoshihide; Mizunoya, Wataru

doi:10.1007/s10974-017-9468-6

Cited by 47 publications

(48 citation statements)

References 31 publications

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“…TAGs, an important energy source, are one of the markers of slow‐twitch fibers because IMCL droplets were more prevalent in the soleus than in EDL tissue (Figure 1, supporting information). Furthermore, Komiya et al have demonstrated that the TAG contents of Type IIa fibers were higher than those of Type I fibers . However, TAGs visualized with fluorescent probes include multiple molecular species.…”

Section: Discussionmentioning

confidence: 99%

Characterization of myofiber‐type‐specific molecules using mass spectrometry imaging

Goto‐Inoue

Morisasa

Machida

et al. 2018

Rapid Comm Mass Spectrometry

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Rationale:In skeletal muscles, there are four myofiber types, Types I, IIa, IIx, and IIb, which show different contraction characteristics and have different metabolic statuses. To understand muscle function, it is necessary to analyze myofiber-specific metabolic changes. However, these fibers are heterogeneous and are hard to discriminate by conventional analyses using tissue extracts. In this study, we found myofiber-specific molecules and molecular markers of other cells such as smooth muscle cells, fat cells, and motor neurons, and visualized them within muscle sections. Methods:We used three different muscle tissues, namely extensor digitorum longus, soleus, and gastrocnemius tissues, from ICR mice. After the muscles had been harvested, cross-sections were prepared using a cryostat and analyzed using matrixassisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI), and conventional immunofluorescence imaging.Results: By comparing the MALDI MSI results with the immunofluorescence imaging results, we were able to identify each fiber and cell-specific ion. It was especially important that we could find Type IIa and IIb specific ions, because these were difficult to distinguish. Conclusions:Through MSI analyses, we performed a comprehensive survey to identify cell-and myofiber-specific molecular markers. In conclusion, we assigned muscle fiber Type I, IIa, and IIb-specific molecular ions at m/z 856.6, 872.6, and 683.8, respectively. These molecular markers might be useful for verifying changes that occur due to exercise and/or disease.

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Section: Discussionmentioning

confidence: 99%

Characterization of myofiber‐type‐specific molecules using mass spectrometry imaging

Goto‐Inoue

Morisasa

Machida

et al. 2018

Rapid Comm Mass Spectrometry

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“…Lipid droplets were stained with BODIPY 493/503 (Life Technologies) diluted in 10% (v/v) dimethyl sulfoxide in PBS to a concentration of 1 mg/mL [17]. Frozen cardiac tissue or NRCMs were fixed with 4% paraformaldehyde for 20 min and then immersed in BODIPY 493/503 solution for 30 min at 37 °C.…”

Section: Bodipy 493/503 Stainingmentioning

confidence: 99%

Exogenous H2S Induces Hrd1 S-sulfhydration and Prevents CD36 Translocation via VAMP3 Ubiquitylation in Diabetic Hearts

Wang

et al. 2020

Aging and disease

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Hydrogen sulfide (H2S) plays physiological roles in vascular tone regulation, cytoprotection, and ATP synthesis. HMG-CoA reductase degradation protein (Hrd1), an E3 ubiquitin ligase, is involved in protein trafficking. H2S may play a role in controlling fatty acid uptake in diabetic cardiomyopathy (DCM) in a manner correlated with modulation of Hrd1 S-sulfhydration; however, this role remains to be elucidated. The aim of the present study was to examine whether H2S can attenuate lipid accumulation and to explain the possible mechanisms involved in the regulation of the H2S-Hrd1/VAMP3 pathway. Db/db mice and neonatal rat cardiomyocytes treated with high glucose, palmitate and oleate were used as animal and cellular models of type 2 diabetes, respectively. The expression of cystathionine-γ-lyase (CSE), Hrd1, CD36 and VAMP3 was detected by Western blot analysis. In addition, Hrd1 was mutated at Cys115, and Hrd1 S-sulfhydration was examined using an S-sulfhydration assay. VAMP3 ubiquitylation was investigated by immunoprecipitation. Lipid droplet formation was tested by TEM, BODIPY 493/503 staining and oil red O staining. The expression of CSE and Hrd1 was decreased in db/db mice compared to control mice, whereas CD36 and VAMP3 expression was increased. NaHS administration reduced droplet formation, and exogenous H2S restored Hrd1 expression, modified Ssulfhydration, and decreased VAMP3 expression in the plasma membrane. Using LC-MS/MS analysis, we identified 85 proteins with decreased ubiquitylation, including 3 vesicle-associated membrane proteins, in the cardiac tissues of model db/db mice compared with NaHS-treated db/db mice. Overexpression of Hrd1 mutated at Cys115 diminished VAMP3 ubiquitylation, whereas it increased CD36 and VAMP3 expression and droplet formation. siRNA-mediated Hrd1 deletion increased the expression of CD36 in the cell membrane. These findings suggested that H2S regulates VAMP3 ubiquitylation via Hrd1 S-sulfhydration at Cys115 to prevent CD36 translocation in diabetes.

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“…And EN occupied the largest number of specific PASs that uniquely expressed. Moreover, we performed a further comparative analysis of tissue-specific PASs between SM and EDL in 1 d and adult periods (366 and 278), which were often used as representative muscles dominated by slowtwitch (type I) and fast-twitch (type II) fibers in animal experiments [44]. Gene ontology analysis revealed that these genes containing tissue-specific PASs (p < 0.05) were mainly involved in oxidation-reduction process, cell division, response to drug, apoptotic process, and inflammatory response in terms of molecular function (Figure 4(b)).…”

Section: Tissue Specificity Analysis Of Passmentioning

confidence: 99%

Characterization and Functional Analysis of Polyadenylation Sites in Fast and Slow Muscles

Deng

Zou

et al. 2020

BioMed Research International

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Many increasing documents have proved that alternative polyadenylation (APA) events with different polyadenylation sites (PAS) contribute to posttranscriptional regulation. However, little is known about the detailed molecular features of PASs and its role in porcine fast and slow skeletal muscles through microRNAs (miRNAs) and RNA binding proteins (RBPs). In this study, we combined single-molecule real-time sequencing and Illumina RNA-seq datasets to comprehensively analyze polyadenylation in pigs. We identified a total of 10,334 PASs, of which 8734 were characterized by reference genome annotation. 32.86% of PASassociated genes were determined to have more than one PAS. Further analysis demonstrated that tissue-specific PASs between fast and slow muscles were enriched in skeletal muscle development pathways. In addition, we obtained 1407 target genes regulated by APA events through potential binding 69 miRNAs and 28 RBPs in variable 3′ UTR regions and some are involved in myofiber transformation. Furthermore, the de novo motif search confirmed that the most common usage of canonical motif AAUAAA and three types of PASs may be related to the strength of motifs. In summary, our results provide a useful annotation of PASs for pig transcriptome and suggest that APA may serve as a role in fast and slow muscle development under the regulation of miRNAs and RBPs.

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Mouse soleus (slow) muscle shows greater intramyocellular lipid droplet accumulation than EDL (fast) muscle: fiber type-specific analysis

Cited by 47 publications

References 31 publications

Characterization of myofiber‐type‐specific molecules using mass spectrometry imaging

Characterization of myofiber‐type‐specific molecules using mass spectrometry imaging

Exogenous H2S Induces Hrd1 S-sulfhydration and Prevents CD36 Translocation via VAMP3 Ubiquitylation in Diabetic Hearts

Characterization and Functional Analysis of Polyadenylation Sites in Fast and Slow Muscles

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