Dataset Information

Electric pulse stimulation in human muscle cells

ABSTRACT: Electrical pulse stimulation (EPS) is used to study exercise-related adaptations in muscle cells in vitro. We compared the effects of two EPS protocols: 1. intermittent high-frequency or 2. continuous low-frequency stimulation, on glucose and fatty acid metabolism, muscle fiber-type markers, mitochondrial proteins, myokines and extracellular vesicles (EV) release. Both EPS protocols enhanced post-exercise glycogen synthesis, while an increase in oxidative capacity was observed only after the intermittent high-frequency stimulation. Both EPS protocols increased GDF11 protein and release of EVs, which were characterized by transmission electron microscopy, nanoparticle tracking analysis and proteomics.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Muscle

SUBMITTER: Václav Pustka

LAB HEAD: Zbynek Zdrahal

PROVIDER: PXD059967 | Pride | 2026-01-12

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	6260_TIMS1_RSLC-110m_TFA_DIA_BC1_1_GA2_1_46797.d.zip	Other
	6260_TIMS1_RSLC-110m_TFA_DIA_BE1_1_GA1_1_46805.d.zip	Other
	6260_TIMS1_RSLC-110m_TFA_DIA_SC1_1_GA4_1_46765.d.zip	Other
	6260_TIMS1_RSLC-110m_TFA_DIA_SC2_1_GA6_1_46789.d.zip	Other
	6260_TIMS1_RSLC-110m_TFA_DIA_SE1_1_GA3_1_46773.d.zip	Other

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Publications

Electrical pulse stimulation reflecting the episodic nature of real-life exercise modulates metabolic and secretory profile of primary human myotubes.

Gabrišová Klára K Kurdiová Tímea T Barkova Daria D Pálešová Natália N Babulicová Jana J Tyčiaková Silvia S Novotová Marta M Balážová Mária M Sabo Miroslav M Pustka Václav V Ukropec Jozef J Ukropcová Barbara B

FEBS open bio 20250909 1

Electrical pulse stimulation (EPS) represents a useful tool to study exercise-related adaptations of muscle cells in vitro. Here, we examine the metabolic and secretory response of primary human muscle cells from metabolically healthy individuals to the EPS protocol reflecting the episodic nature of real-life exercise training. This intermittent EPS protocol alternates high-frequency stimulation periods with low-frequency resting periods. Continuous EPS was used as a comparator. Radiometric asse ...[more]

PMID: 40923494

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Project description:Regular physical exercise evokes profound physiological responses which are strongly associated with many health benefits. However, the details of cellular networks and mechanisms underlying the adaptive responses to exercise remain to be elucidated. We have previously shown the usefulness of electrical pulse stimulation (EPS) to investigate metabolic effects of exercise in cultured human myotubes. The aim of the present study was to uncover networks of signaling pathways and regulatory molecules responsible for the metabolic effects of exercise in human skeletal muscle cells exposed to chronic EPS. Differentiated myotubes were subjected to two different EPS protocols (protocol 1; 2 ms, 10 V, 0.1 Hz for 24 h or protocol 2; 2 ms, 30 V, and 1 Hz for 48 h). Fuel handling was assessed using radiolabeled substrates. The transcriptome, cell proteome, and secreted proteins from EPS-treated and untreated myotubes were analyzed using a combination of high-throughput RNA sequencing, microarray, and high-resolution liquid chromatography mass spectrometry. Independent validation of selected putative myokines were measured using ELISA or multiplex assay. Oxidative metabolism was enhanced in human myotubes exposed to EPS protocol 1. A total of 81 differentially regulated proteins and 952 differentially expressed genes (DEGs) were observed in the cells after EPS protocol 1. Gene ontology (GO) analysis indicated that significantly regulated proteins and genes were enriched in biological processes related to glycolytic pathways, positive regulation of fatty acid oxidation, and oxidative phosphorylation, as well as muscle contraction, autophagy/mitophagy, and oxidative stress. Moreover, proteomic identification of secreted proteins revealed extracellular levels of 137 proteins were changed in myotubes subjected to EPS protocol 1. We also found some degree of overlap between the DEGs found in myotubes submitted to EPS protocol 1 and protocol 2. Among these DEGs was a myokine, leukemia inhibitory factor, which showed to enhance the glucose uptake in skeletal muscle cells, indicating autocrine mechanism to maintain metabolic homeostasis. Collectively, our data provides new insight into the transcriptome, proteome and secreted proteins alterations following in vitro exercise and is a valuable resource for understanding the molecular mechanisms and regulatory molecules mediating the beneficial metabolic effects of exercise.

Project description:Lifestyle disorders like obesity, type 2 diabetes (T2D), and cardiovascular diseases can be prevented and treated by regular physical activity. During exercise, skeletal muscles release signaling factors that communicate with other organs and mediate beneficial effects of exercise. These factors include myokines, metabolites, and extracellular vesicles (EVs). In the present study, we have examined how electrical pulse stimulation (EPS) of myotubes, a model of exercise, affects the cargo of released EVs. Chronic low frequency EPS was applied for 24 h to human myotubes isolated and differentiated from biopsy samples from 6 morbidly obese females with T2D, and EVs, both exosomes and microvesicles (MV), were isolated from cell media 24 h thereafter. Size and concentration of EV subtypes were characterized by nanoparticle tracking analysis, surface markers were examined by flow cytometry and Western blotting, and morphology was confirmed by transmission electron microscopy. Protein content was assessed by high-resolution proteomic analysis (LC-MS/MS), non-coding RNA was quantified by Affymetrix microarray, and selected microRNAs (miRs) validated by real time RT-qPCR. The size and concentration of exosomes and MV were unaffected by EPS. Of the 400 miRs identified in the EVs, EPS significantly changed the level of 15 exosome miRs, of which miR-1233-5p showed the highest fold change. The miR pattern of MV was unaffected by EPS. Totally, about 1000 proteins were identified in exosomes and 2000 in MV. EPS changed the content of 73 proteins in exosomes, 97 in MVs, and of these 4 were changed in both exosomes and MV (GANAB, HSPA9, CNDP2, and ATP5B). By matching the EPS-changed miRs and proteins in exosomes, 31 targets were identified, and among these several promising signaling factors. Of particular interest were CNDP2, an enzyme that generates the appetite regulatory metabolite Lac-Phe, and miR-4433b-3p, which targets CNDP2. Several of the regulated miRs, such as miR-92b-5p, miR-320b, and miR-1233-5p might also mediate interesting signaling functions.