Skeletal muscle ribosome analysis: a comparison of common assay methods and utilization of a novel RiboAb antibody cocktail DOI Creative Commons
Joshua S. Godwin, J. Max Michel, C. Brooks Mobley

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 9, 2024

ABSTRACT Cellular and tissue total RNA concentrations have been widely reported to represent ribosome content, a metric that reflects the trophic state of skeletal muscle. Although various assays are used assess concentrations, there is need homologize quantification approaches. Thus, we analyzed C2C12 myotubes mouse muscle determine if provided through UV-Vis spectroscopy (UV), fluorometry only (Fluor), fluorometry-based microfluidic chip electrophoresis (MFGE) were representative cellular rRNA (i.e., MFGE 18S+28S rRNA, criterion content). We also sought whether novel ribosomal protein antibody cocktail (termed RiboAb ) corresponded with concentrations. Compared non-treated myotubes, 24-hour insulin-like growth factor-1 (IGF-1) treatments increased (∼2.0-fold; p<0.001) based on UV (∼1.9-fold; p<0.001), Fluor (∼2.3 fold; p=0.001), (∼2.1-fold, p<0.001). In C57BL/6 mice, 10 days mechanical overload (MOV) via synergist ablation elevated plantaris (∼1.7-fold; p=0.017) according (∼1.5-fold; p=0.033), (∼1.6-fold; (∼1.8-fold, p=0.017). both experiments, concentration data yielded from all three techniques exhibited significant positive correlations Ribosome pelleting experiments indicated proteins assayed (rps3/6 rpl5/11) exclusively associated pellet. Additionally, myotube levels higher IGF-1 MOV, respectively, relative controls (1.3-fold 1.7-fold, p<0.017), values correlated (r=0.637 r=0.853, p<0.005). These confirm UV, Fluor, valid surrogates cell/tissue content. propose may serve as surrogate for changes in content these models, although future research needed examine feasibility humans well utility other applications (e.g., immunohistochemistry and/or fractionation experiments).

Language: Английский

Microvascular Skeletal-Muscle Crosstalk in Health and Disease DOI Open Access
Gerald J. Pepe, Eugene D. Albrecht

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(13), P. 10425 - 10425

Published: June 21, 2023

As an organ system, skeletal muscle is essential for the generation of energy that underpins contraction, plays a critical role in controlling balance and insulin-dependent glucose homeostasis, as well vascular well-being, regenerates following injury. To achieve there requirement "cross-talk" between myogenic components their regulatory factors comprise muscle. Accordingly, this review will describe following: [a] embryonic cell-signaling events important establishing cell-lineage, cross-talk endothelial cells (EC) precursors underpinning development muscle, its vasculature satellite-stem-cell (SC) pool, EC-SC maintains SC quiescence localizes ECs to SCs angio-myogenesis postnatally; [b] vascular-myocyte actions insulin on vasodilation capillary surface area uptake glucose/insulin by myofibers microvascular-myocyte dysfunction characterizes resistance, diabetes hypertension, estrogen growth adults; [c] utero fetal skeletal-muscle microvascularization myofiber hypertrophy required metabolic/vascular homeostasis after birth; [d] interactions underpin regeneration post-injury; [e] Duchenne muscular dystrophy.

Language: Английский

Citations

9
Advances in tissue engineering approaches for repairing and rehabilitating the myotendinous junction DOI

Kariman A. Shama,

Mariah A. Turner,

Harrison B. Broadaway

et al.

Current Opinion in Biomedical Engineering, Journal Year: 2024, Volume and Issue: 30, P. 100532 - 100532

Published: March 29, 2024

Language: Английский

Citations

2
Differential impact of sex on regulation of skeletal muscle mitochondrial function and protein homeostasis by hypoxia‐inducible factor‐1α in normoxia DOI
Nicole Welch, Saurabh Mishra,

Annette Bellar

et al.

The Journal of Physiology, Journal Year: 2024, Volume and Issue: 602(12), P. 2763 - 2806

Published: May 18, 2024

Hypoxia-inducible factor (HIF)-1α is continuously synthesized and degraded in normoxia. During hypoxia, HIF1α stabilization restricts cellular/mitochondrial oxygen utilization. Cellular stressors can stabilize even during However, less known about function(s) sex-specific effects normoxia the basal state. Since skeletal muscle largest protein store mammals homeostasis has high energy demands, we determined function at baseline muscle. Untargeted multiomics data analyses were followed by experimental validation differentiated murine myotubes with loss/gain of from mice without/with post-natal muscle-specific Hif1a deletion (Hif1a

Language: Английский

Citations

2
Does sex matter in the cheetah? Insights into the skeletal muscle of the fastest land animal DOI Creative Commons
Tertius A. Kohn,

Samantha Knobel,

Byron Donaldson

et al.

Journal of Experimental Biology, Journal Year: 2024, Volume and Issue: 227(15)

Published: July 18, 2024

The cheetah is considered the fastest land animal, but studies on their skeletal muscle properties are scarce. Vastus lateralis biopsies, obtained from male and female cheetahs as well humans, were analysed compared for fibre type size, metabolism. Overall, had predominantly IIX fibres, which was confirmed by myosin heavy chain isoform content (mean±s.d. I: 17±8%, IIA: 21±6%, IIX: 62±12%), whereas human contained I IIA fibres (type 49±14%, 43±8%, 7±7%). Cheetahs smaller than with larger in males counterparts. Citrate synthase (16±6 versus 28±7 µmol min-1 g-1 protein, P<0.05) 3-hydroxyacyl co-enzyme A dehydrogenase (30±11 47±15 activities lower lactate activity 6 times higher (2159±827 382±161 P<0.001). of creatine kinase (4765±1828 6485±1298, P<0.05 protein) phosphorylase (111±29 216±92 irrespective cheetahs. Superoxide dismutase catalase, markers antioxidant capacity, overall capacity To conclude, type, size metabolism differ between limited differences sexes.

Language: Английский

Citations

2
Muscle miRNAs are influenced by sex at baseline and in response to exercise DOI Creative Commons
Danielle Hiam, Shanie Landen, Macsue Jacques

et al.

BMC Biology, Journal Year: 2023, Volume and Issue: 21(1)

Published: Nov. 27, 2023

Language: Английский

Citations

6
Metabolic dysregulation and decreased capillarization in skeletal muscles of male adolescent offspring rats exposed to gestational intermittent hypoxia DOI Creative Commons

Wirongrong Wongkitikamjorn,

E. Wada, Jun Hosomichi

et al.

Frontiers in Physiology, Journal Year: 2023, Volume and Issue: 14

Published: Jan. 12, 2023

Gestational intermittent hypoxia (IH) is a hallmark of obstructive sleep apnea that occurs frequently during pregnancy, and effects caused by this environmental change pregnancy may be transmitted to the offspring. In study, we aimed clarify IH in pregnant rats on skeletal muscle adolescent offspring rats. Mother underwent from gestation day 7-21, their 5-weeks-old male were analyzed. All born raised under normoxia conditions. Although no general growth retardation was observed, found exposure gestational reduces endurance running capacity Both respiratory (diaphragm; DIA) limb (tibialis anterior; TA) showed histological abnormalities, including fiber size type distribution. To identify possible mechanism underlying reduced capacity, regulatory factors associated with energy metabolism analyzed different parts muscles. Compared conditions normoxia, significantly lower expression genes glucose lipid metabolism, protein levels phosphorylated AMPK AKT. Furthermore, gene adiponectin receptors one two decreased DIA TA addition, had capillary density as result IH. However, these changes not observed sucking (geniohyoid) masticating (masseter) These results suggest muscles are vulnerable IH, which induces altered aerobic motor function. partially owing

Language: Английский

Citations

4
Lactobacillus gasseri BNR17 Ameliorates Dexamethasone-Induced Muscle Loss in BALB/c Mice and C2C12 Myotubes DOI

Hyeon-Yeong Lee,

Jong‐Kyu Lee,

Hyemi Lim

et al.

Journal of Medicinal Food, Journal Year: 2024, Volume and Issue: 27(5), P. 385 - 395

Published: April 4, 2024

This study aimed to investigate the effects and mechanism of

Language: Английский

Citations

1
Have a little heart (or not): highly minimized skeletal muscle regulatory cassettes with low or no activity in heart DOI
Charis L. Himeda, Takako I. Jones, Peter L. Jones

et al.

Human Gene Therapy, Journal Year: 2024, Volume and Issue: 35(15-16), P. 543 - 554

Published: Aug. 1, 2024

Adeno-associated virus-mediated gene therapies for certain muscle disorders require regulatory cassettes that provide high-level, striated muscle-specific activity. However, cardiotoxicity has emerged as a serious concern in clinical trials Duchenne muscular dystrophy and X-linked myotubular myopathy. While this may be caused by systemic inflammatory effects of the treatment, high transgene expression heart also play role. Thus, modulated level therapeutic heart, while others not any cardiac at all. Additionally, size some cargos requires to small enough large cDNAs other payloads can accommodated. we have performed enhancer/promoter optimization develop highly minimized are active skeletal muscles, with either low or no detectable activity muscle. Our No-heart (NH) cassette is most but exhibits only very extensor digitorum longus (EDL), soleus, diaphragm, heart. By contrast, our Have Little Heart (HLH) displays comparable ∼800-bp CK8 cassette, increased EDL, Due their size, these used strategies both flexible (

Language: Английский

Citations

1
Switching ubiquitous and muscle‐specific isoforms of mitochondrial respiratory complex IV in skeletal muscle fine‐tunes complex IV activity DOI
Xianglai Ye,

Yaojun Xie,

Yu Shi

et al.

The FASEB Journal, Journal Year: 2023, Volume and Issue: 37(4)

Published: March 24, 2023

Abstract Respiratory complex IV (CIV, cytochrome c oxidase) is the terminal enzyme of mitochondrial electron transport chain. Some CIV subunits have two or more isoforms, which are ubiquitously expressed in specific tissues like lung, muscle, and testis. Among tissue‐specific muscle‐specific isoforms adult cardiac skeletal muscles. To date, physiological biochemical association between aerobic respiration muscles remains unclear. In this study, we profiled organization expression pattern different mouse muscle tissues. We found extensive CIV‐containing supramolecular murine musculature at advanced developmental stages, while a switch from ubiquitous to was also detected. Such confirmed during vitro differentiation C2C12 myoblasts. Unexpectedly, decrease observed differentiation, probably due small increase coupled with dramatic isoforms. that enzymatic activity containing isoform COX6A2 higher than COX6A1 engineered HEK293T cells. Overall, our results indicate switching indispensable for optimized oxidative phosphorylation mature note model not suitable study muscular insufficient

Language: Английский

Citations

2
Different responses of skeletal muscles to femoral artery ligation-induced ischemia identified in BABL/c and C57BL/6 mice DOI Creative Commons

Huiyin Tu,

Junliang Qian, Dongze Zhang

et al.

Frontiers in Physiology, Journal Year: 2022, Volume and Issue: 13

Published: Sept. 16, 2022

Peripheral arterial disease (PAD) is a common circulatory problem in lower extremities, and the murine ischemic model used to reproduce human PAD. To compare strain differences of skeletal muscle responses ischemia, left femoral artery was blocked by ligation reduce blood flow limb BALB/c C57BL/6 mice. After 6 weeks ligation, functional morphological changes gastrocnemius were evaluated. mice displayed serious muscular dystrophy, including smaller myofibers (524.3 ± 66 µM2), accumulation adipose-liked tissue (17.8 0.9%), fibrosis (6.0 0.5%), compared (1,328.3 76.3 µM2, 0.27 0.09%, 1.56 0.06%, respectively; p < 0.05). About neuromuscular junctions (NMJs) muscle, induced more damage than that mice, demonstrated fragment number nicotinic acetylcholine receptor (nAChR) clusters (8.8 1.3 vs. 2.5 0.7 0.05) amplitude sciatic nerve stimulated-endplate potentials (EPPs) (9.29 1.34 mV 20.28 1.42 More importantly, significantly weakened nerve-stimulated contraction whereas it didn't alter These results suggest useful animal develop new therapeutic approaches improve structure function PAD, although mechanisms about ischemia are unclear.

Language: Английский

Citations

4