electrical pulse stimulation
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Author(s):  
Alec Brendan Chaves ◽  
Edwin R. Miranda ◽  
Jacob T. Mey ◽  
Brian K. Blackburn ◽  
Kelly N.Z Fuller ◽  
...  

Thioredoxin-interacting protein (TXNIP) negatively effects the redox state and growth signaling via its interactions with thioredoxin (TRX) and regulated in development and DNA damage response 1 (REDD1), respectively. TXNIP expression is downregulated by pathways activated during aerobic exercise (AE), via posttranslational modifications (PTMs; serine phosphorylation and ubiquitination). The purpose of this investigation was to determine the effects of acute AE on TXNIP expression, posttranslational modifications, and its interacting partners, REDD1 and TRX. Fifteen healthy adults performed 30 minutes of aerobic exercise (80% VO2max) with muscle biopsies taken before, immediately following, and three hours following the exercise bout. To explore potential mechanisms underlying our in vivo findings, primary human myotubes were exposed to two models of exercise, electrical pulse stimulation (EPS) and palmitate-forskolin-ionomycin (PFI). Immediately following exercise, TXNIP protein decreased, but returned to pre-exercise levels three hours post exercise. These results were replicated in our PFI exercise model only. Although not statistically significant, there was a trending main effect in serine-phosphorylation status of TXNIP (p=0.07) immediately following exercise. REDD1 protein decreased three hours after exercise. AE had no effect on TRX protein expression, gene expression or the activity of its reducing enzyme, thioredoxin reductase. Consequently, AE had no effect on the TRX: TXNIP interaction. Our results indicate that AE leads to acute reductions in TXNIP and REDD1 protein expression. However, these changes did not result in alterations in the TRX: TXNIP interaction and could not be entirely explained by alterations in TXNIP PTMs or changes in TRX expression or activity.



Life ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1227
Author(s):  
Eleni Nintou ◽  
Eleni Karligiotou ◽  
Maria Vliora ◽  
Ioannis G. Fatouros ◽  
Athanasios Z. Jamurtas ◽  
...  

The crosstalk between the exercising muscle and the adipose tissue, mediated by myokines and metabolites, derived from both tissues during exercise has created a controversy between animal and human studies with respect to the impact of exercise on the browning process. The aim of this study was to investigate whether co-culturing of C2C12 myotubes and 3T3-L1 adipocytes under the stimuli of electrical pulse stimulation (EPS) mimicking muscle contraction can impact the expression of UCP1, PGC-1a, and IL-6 in adipocytes, therefore providing evidence on the direct crosstalk between adipocytes and stimulated muscle cells. In the co-cultured C2C12 cells, EPS increased the expression of PGC-1a (p = 0.129; d = 0.73) and IL-6 (p = 0.09; d = 1.13) protein levels. When EPS was applied, we found that co-culturing led to increases in UCP1 (p = 0.044; d = 1.29) and IL-6 (p = 0.097; d = 1.13) protein expression in the 3T3-L1 adipocytes. The expression of PGC-1a increased by EPS but was not significantly elevated after co-culturing (p = 0.448; d = 0.08). In vitro co-culturing of C2C12 myotubes and 3T3-L1 adipocytes under the stimuli of EPS leads to increased expression of thermogenic proteins. These findings indicate changes in the expression pattern of proteins related to browning of adipose tissue, supporting the use of this in vitro model to study the crosstalk between adipocytes and contracting muscle.



2021 ◽  
Vol 11 (21) ◽  
pp. 10436
Author(s):  
Taku Fukushima ◽  
Miho Takata ◽  
Ayano Kato ◽  
Takayuki Uchida ◽  
Takeshi Nikawa ◽  
...  

Exercise has beneficial effects on human health and is affected by two different pathways; motoneuron and endocrine. For the advancement of exercise research, in vitro exercise models are essential. We established two in vitro exercise models using C2C12 myotubes; EPS (electrical pulse stimulation) for a motoneuron model and clenbuterol, a specific β2 adrenergic receptor agonist, treatment for an endocrine model. For clenbuterol treatment, we found that Ppargc1a was induced only in low glucose media (1 mg/mL) using a 1-h treatment of 30 ng/mL clenbuterol. Global transcriptional changes of clenbuterol treatment were analyzed by RNA-seq and gene ontology analyses and indicated that mitogenesis and the PI3K-Akt pathway were enhanced, which is consistent with the effects of exercise. Cxcl1 and Cxcl5 were identified as candidate myokines induced by adrenaline. As for the EPS model, we compared 1 Hz of 1-pulse EPS and 1 Hz of 10-pulse EPS for 24 h and determined Myh gene expressions. Ten-pulse EPS induced higher Myh2 and Myh7 expression. Global transcriptional changes of 10-pulse EPS were also analyzed using RNA-seq, and gene ontology analyses indicated that CaMK signaling and hypertrophy pathways were enhanced, which is also consistent with the effects of exercise. In this paper, we provided two transcriptome results of in vitro exercise models and these databases will contribute to advances in exercise research.



Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3002
Author(s):  
Matthias Catteau ◽  
Emilie Passerieux ◽  
Léo Blervaque ◽  
Farés Gouzi ◽  
Bronia Ayoub ◽  
...  

Among the comorbidities associated with chronic obstructive pulmonary disease (COPD), skeletal muscle weakness and atrophy are known to affect patient survival rate. In addition to muscle deconditioning, various systemic and intrinsic factors have been implicated in COPD muscle dysfunction but an impaired COPD muscle adaptation to contraction has never been extensively studied. We submitted cultured myotubes from nine healthy subjects and nine patients with COPD to an endurance-type protocol of electrical pulse stimulation (EPS). EPS induced a decrease in the diameter, covered surface and expression of MHC1 in COPD myotubes. Although the expression of protein degradation markers was not affected, expression of the protein synthesis marker mTOR was not induced in COPD compared to healthy myotubes after EPS. The expression of the differentiation markers p16INK4a and p21 was impaired, while expression of Myf5 and MyoD tended to be affected in COPD muscle cells in response to EPS. The expression of mitochondrial biogenesis markers PGC1α and MFN2 was affected and expression of TFAM and COX1 tended to be reduced in COPD compared to healthy myotubes upon EPS. Lipid peroxidation was increased and the expression of the antioxidant enzymes SOD2 and GPx4 was affected in COPD compared to healthy myotubes in response to EPS. Thus, we provide evidence of an impaired response of COPD muscle cells to contraction, which might be involved in the muscle weakness observed in patients with COPD.



Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2556
Author(s):  
Kantaro Yoshioka ◽  
Akira Ito ◽  
Masanobu Horie ◽  
Kazushi Ikeda ◽  
Sho Kataoka ◽  
...  

Duchenne muscular dystrophy (DMD) is a genetic disorder that results from deficiency of the dystrophin protein. In recent years, DMD pathological models have been created using induced pluripotent stem (iPS) cells derived from DMD patients. In addition, gene therapy using CRISPR-Cas9 technology to repair the dystrophin gene has been proposed as a new treatment method for DMD. However, it is not known whether the contractile function of myotubes derived from gene-repaired iPS cells can be restored. We therefore investigated the maturation of myotubes in electrical pulse stimulation culture and examined the effect of gene repair by observing the contractile behaviour of myotubes. The contraction activity of myotubes derived from dystrophin-gene repaired iPS cells was improved by electrical pulse stimulation culture. The iPS cell method used in this study for evaluating muscle contractile activity is a useful technique for analysing the mechanism of hereditary muscular disease pathogenesis and for evaluating the efficacy of new drugs and gene therapy.



PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247377
Author(s):  
Vid Jan ◽  
Katarina Miš ◽  
Natasa Nikolic ◽  
Klemen Dolinar ◽  
Metka Petrič ◽  
...  

Denervation reduces the abundance of Na+,K+-ATPase (NKA) in skeletal muscle, while reinnervation increases it. Primary human skeletal muscle cells, the most widely used model to study human skeletal muscle in vitro, are usually cultured as myoblasts or myotubes without neurons and typically do not contract spontaneously, which might affect their ability to express and regulate NKA. We determined how differentiation, de novo innervation, and electrical pulse stimulation affect expression of NKA (α and β) subunits and NKA regulators FXYD1 (phospholemman) and FXYD5 (dysadherin). Differentiation of myoblasts into myotubes under low serum conditions increased expression of myogenic markers CD56 (NCAM1), desmin, myosin heavy chains, dihydropyridine receptor subunit α1S, and SERCA2 as well as NKAα2 and FXYD1, while it decreased expression of FXYD5 mRNA. Myotubes, which were innervated de novo by motor neurons in co-culture with the embryonic rat spinal cord explants, started to contract spontaneously within 7–10 days. A short-term co-culture (10–11 days) promoted mRNA expression of myokines, such as IL-6, IL-7, IL-8, and IL-15, but did not affect mRNA expression of NKA, FXYDs, or myokines, such as musclin, cathepsin B, meteorin-like protein, or SPARC. A long-term co-culture (21 days) increased the protein abundance of NKAα1, NKAα2, FXYD1, and phospho-FXYD1Ser68 without attendant changes in mRNA levels. Suppression of neuromuscular transmission with α-bungarotoxin or tubocurarine for 24 h did not alter NKA or FXYD mRNA expression. Electrical pulse stimulation (48 h) of non-innervated myotubes promoted mRNA expression of NKAβ2, NKAβ3, FXYD1, and FXYD5. In conclusion, low serum concentration promotes NKAα2 and FXYD1 expression, while de novo innervation is not essential for upregulation of NKAα2 and FXYD1 mRNA in cultured myotubes. Finally, although innervation and EPS both stimulate contractions of myotubes, they exert distinct effects on the expression of NKA and FXYDs.



2021 ◽  
Vol 7 ◽  
Author(s):  
Pim G. Boute ◽  
Maarten Soetaert ◽  
José A. Reid Navarro ◽  
Martin J. Lankheet

Electrical pulse trawling is an alternative to conventional beam trawling for common sole (Solea solea), with the potential for higher revenues and less impact on the marine ecosystem. Concerns exist, however, that benthic invertebrates might be seriously affected by pulse fishing. Even if direct injuries and mortality were limited, changes in behaviour might compromise their survival, with potentially large impacts on food webs. Here, we investigate effects of electrical pulses on locomotion behaviour and 14-days survival of six invertebrate species from four phyla that may encounter pulse fishing gears. Electrical stimulation consisted of a Pulsed Bipolar Current at 200 V m–1, 30 Hz, 0.33 ms pulse width, and 3 s duration. We quantified species-specific behaviours before, during, and after electrical stimulation and compared these to a non-exposed control group. Responses during stimulation varied from no visible effect (echinoderms) to squirming (sea mouse) and retractions (whelk and crustaceans). Within 30 s after stimulation, all animals resumed normal behavioural patterns, without signs of lasting immobilisation. Starfish, serpent star, whelk and sea mouse showed no change in movement patterns after stimulation, whereas flying crab and hermit crab showed significant changes in activity that were indicative of increased shelter behaviour. For none of the species, survival at 14-days after stimulation was negatively affected. These findings suggest that changes in locomotion behaviour due to electrical stimulation as used in pulse trawling are unlikely to substantially compromise survival of the investigated species.



2021 ◽  
Vol 35 (11) ◽  
pp. 1326-1327
Author(s):  
Somen Baidya ◽  
Ahmed Hassan

Exposing cells to ultrashort electric field pulses can lead to the permeabilization of the membranes of the internal cell organelles, such as the nucleus, which is termed as supra-electroporation. In this paper, we study the supra-electroporation of stem cells with realistic morphologies under nanosecond electrical pulse stimulation. For such short pulses, the cell and the nucleus membranes exhibit non-linearity in their conductance. Therefore, we used a non-linear model for the cells’ membranes, coupled with a quasi-static electromagnetic solver, and obtained the solution using a commercial Finite Element Method (FEM) solver. The results show that the outer shape of the cell has a strong effect on the magnitude and the spatiotemporal patterns of the electric field inside the cell, which affects the rate of the supra-electroporation of the nucleus. These variations in the rate of the supra-electroporation of the nucleus can guide the selective targeting of desired cells with specific shapes.



2020 ◽  
Vol 28 (1) ◽  
pp. 7-11
Author(s):  
Michael J. Eckert ◽  
Kartik Iyer ◽  
David R. Euston ◽  
Masami Tatsuno


2020 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Benjamin A. Kugler ◽  
Wenqian Deng ◽  
Bergomi Francois ◽  
Meaghan Nasta ◽  
J. Matthew Hinkley ◽  
...  


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