Methods to Assess Subcellular Compartments of Muscle in &lt;em&gt;C. elegans&lt;/em&gt;

Gaffney, Christopher; Bass, J.; Barratt, Thomas F.; Szewczyk, Nathaniel J.

doi:10.3791/52043-v

Cited by 2 publications

(2 citation statements)

References 16 publications

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“…To assess protein degradation, the animals were stained with X-gal. 30 Briefly, the worms were washed down onto slides and dried using a dryer for 30 minutes, then the slides were submerged in ice-cold acetone for 3.5 min and stained with X-gal solution for 2 hours in a humid environment, and then images were obtained using a fluorescence microscope (Nikon, Eclipse Ti–S). The staining area was analyzed manually using ImageJ software ( n > 30, three independent trials).…”

Section: Methodsmentioning

confidence: 99%

Linolenic acid ameliorates sarcopenia in C. elegans by promoting mitophagy and fighting oxidative stress

Zhang

et al. 2023

Food Funct.

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Section: Methodsmentioning

confidence: 99%

Linolenic acid ameliorates sarcopenia in C. elegans by promoting mitophagy and fighting oxidative stress

Zhang

et al. 2023

Food Funct.

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“…The video was then analysed using ImageJ and wrMTrck (wrMTrck multiple object tracker ‐ phage.dk). 13 …”

Section: Methodsmentioning

confidence: 99%

Development of a traditional Chinese medicine‐based agent for the treatment of cancer cachexia

Chu

Cheng

et al. 2022

J cachexia sarcopenia muscle

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Background Despite recent advances in understanding the pathophysiology of cancer cachexia, prevention/treatment of this debilitating disease remains an unmet medical need. Methods We developed an integrated, multi‐tiered strategy involving both in vitro and in vivo muscle atrophy platforms to identify traditional Chinese medicine (TCM)‐based anti‐cachectic agents. In the initial screening, we used inflammatory cytokine‐induced atrophy of C2C12 myotubes as a phenotypic screening platform to assess the protective effects of TCMs. The selected TCMs were then evaluated for their abilities to protect Caenorhabditis elegans from age‐related reduction of mobility and contractility, followed by the C‐26 colon adenocarcinoma mouse model of cachexia to confirm the anti‐muscle atrophy effects (body/skeletal muscle weights, fibre size distribution, grip strengths, and serum IL‐6). Transcriptome analysis, quantitative real‐time polymerase chain reaction, and immunoblotting were performed to gain understanding of the potential mechanism(s) by which effective TCM protected against C26 tumour‐induced muscle atrophy. Results Of 29 widely used TCMs, Dioscorea radix (DR) and Mu Dan Pi (MDP) showed a complete protection (all P values, 0.0002) vis‐à‐vis C26 conditioned medium control in the myotube atrophy platform. MDP exhibited a unique ability to ameliorate age‐associated decreases in worm mobility, accompanied by improved total body contractions, relative to control (P < 0.0001 and <0.01, respectively), which, however, was not noted with DR. This differential in vivo protective effect between MDP and DR was also confirmed in the C‐26 mouse model. MDP at 1000 mg/kg (MDP‐H) was effective in protecting body weight loss (P < 0.05) in C‐26 tumour‐bearing mice without changing food or water intake, accompanied by the restoration of the fibre size distribution of hindleg skeletal muscles (P < 0.0001) and the forelimb grip strength (P < 0.05). MDP‐treated C‐26‐tumour‐bearing mice were alert, showed normal posture and better body conditions, and exhibited lower serum IL‐6 levels (P = 0.06) relative to vehicle control. This decreased serum IL‐6 was associated with the in vitro suppressive effect of MDP (25 and 50 μg/mL) on IL‐6 secretion into culture medium by C26 cells. RNA‐seq analysis, followed by quantitative real‐time polymerase chain reaction and/or immunoblotting, shows that MDP's anti‐cachectic effect was attributable to its ability to reverse the C‐26 tumour‐induced re‐programming of muscle homoeostasis‐associated gene expression, including that of two cachexia drivers (MuRF1 and Atrogin‐1), in skeletal muscles. Conclusions All these findings suggest the translational potential of MDP to foster new strategies for the prevention and/or treatment of cachexia. The protective effect of MDP on other types of muscle atrophy such as sarcopenia might warrant investigations.

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Methods to Assess Subcellular Compartments of Muscle in <em>C. elegans</em>

Cited by 2 publications

References 16 publications

Linolenic acid ameliorates sarcopenia in C. elegans by promoting mitophagy and fighting oxidative stress

Linolenic acid ameliorates sarcopenia in C. elegans by promoting mitophagy and fighting oxidative stress

Development of a traditional Chinese medicine‐based agent for the treatment of cancer cachexia

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