Skip to main content
SpringerLink
Log in

Extracellular stimulation with human “noisy” electromyographic patterns facilitates myotube activity

  • Original Paper
  • Published:
Journal of Muscle Research and Cell Motility Aims and scope Submit manuscript

Abstract

Electrical stimulation (ES) of skeletal muscle partially mimics the benefits of physical activity. However, the stimulation protocols applied clinically to date, often cause unpleasant symptoms and muscle fatigue. Here, we compared the efficiency of a “noisy” stimulus waveform derived from human electromyographic (EMG) muscle patterns, with stereotyped 45 and 1 Hz electrical stimulations applied to mouse myotubes in vitro. Human gastrocnemius medialis electromyograms recorded from volunteers during real locomotor activity were used as a template for a noisy stimulation, called EMGstim. The stimulus-induced electrical activity, intracellular Ca2+ dynamics and mechanical twitches in the myotubes were assessed using whole-cell perforated patch-clamp, Ca2+ imaging and optical visualization techniques. EMGstim was more efficient in inducing myotube cell firing, [Ca2+]i changes and contractions compared with more conventional electrical stimulation. Its stimulation strength was also much lower than the minimum required to induce contractions via stereotyped stimulation protocols. We conclude that muscle cells in vitro can be more efficiently depolarized using the “noisy” stochastic stimulation pattern, EMGstim, a finding that suggests a way to favor a higher level of electrical activity in a larger number of cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Agarwal S, Kobetic R, Nandurkar S, Marsolais EB (2003) Functional electrical stimulation for walking in paraplegia: 17-year follow-up of 2 cases. J Spinal Cord Med 26:86–91

    PubMed  Google Scholar 

  • Bergquist AJ, Clair JM, Lagerquist O, MangCS Okuma Y, Collins DF (2011) Neuromuscular electrical stimulation: implications of the electrically evoked sensory volley. Eur J Appl Physiol 111:2409–2426

    Article  CAS  PubMed  Google Scholar 

  • Bickel CS, Slade JM, Warren GL, Dudley GA (2003) Fatigability and variable-frequency train stimulation of human skeletal muscles. Phys Ther 83:366–373

    PubMed  Google Scholar 

  • Bickel CS, Gregory CM, Dean JC (2011) Motor unit recruitment during neuromuscular electrical stimulation: a critical appraisal. Eur J Appl Physiol 111:2399–2407

    Article  PubMed  Google Scholar 

  • Burch N, Arnold AS, Item F, Summermatter S, Brochmann Santana Santos G, Christe M et al (2010) Electric pulse stimulation of cultured murine muscle cells reproduces gene expression changes of trained mouse muscle. PLoS ONE 5:e10970

    Article  PubMed Central  PubMed  Google Scholar 

  • Buvinic S, Almarza G, Bustamante M, Casas M, Lopez J, Riquelme M et al (2009) ATP released by electrical stimuli elicits calcium transients and gene expression in skeletal muscle. J Biol Chem 284:34490–34505

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Clausen T (2013) Excitation-induced exchange of Na+, K+, Cl− in rat EDL muscle in vitro and in vivo: physiology and pathology. J Gen Physiol 141:179–192

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Delitto A, Strube MJ, Shulman AD, Minor SD (1992) A study of discomfort with electrical stimulation. Phys Ther 72:410–421

    CAS  PubMed  Google Scholar 

  • Dose F, Taccola G (2012) Co-application noisy patterned electrical stimuli and NMDA plus serotonin facilitates fictive locomotion in the rat spinal cord. J Neurophysiol 108:2977–2990

    Article  CAS  PubMed  Google Scholar 

  • Dose F, Menosso R, Taccola G (2013) Rat locomotor spinal circuits in vitro are activated by electrical stimulation with noisy waveforms sampled from human gait. Physiol Rep 1:1–15

    Article  Google Scholar 

  • Downey RJ, Tate M, Kawai H, Dixon WE (2014) Comparing the force ripple during asynchronous and conventional stimulation. Muscle Nerve. doi:10.1002/mus.24186

    PubMed  Google Scholar 

  • Eberstein A, Eberstein S (1996) Electrical stimulation of denervated muscle: is it worthwhile? Med Sci Sports Exerc 8:1463–1469

    Article  Google Scholar 

  • Gomis M, González LM, Querol F, Gallach JE, Toca-Herrera JL (2009) Effects of electrical stimulation on muscle trophism in patients with hemophilic arthropathy. Arch Phys Med Rehabil 90:1924–1930

    Article  PubMed  Google Scholar 

  • Gregory CM, Bickel CS (2005) Recruitment patterns in human skeletal muscle during electrical stimulation. Phys Ther 85:358–364

    PubMed  Google Scholar 

  • Gregory CM, Gregory CM, Bickel CS (2005) Recruitment patterns in human skeletal muscle during electrical stimulation. Phys Ther 85:358–364

    PubMed  Google Scholar 

  • Hayashibe M, Zhang Q, Guiraud D, Fattal C (2011) Evoked EMG-based torque prediction under muscle fatigue in implanted neural stimulation. J Neural Eng 8:064001

    Article  PubMed  Google Scholar 

  • Irintchev A, Langer M, Zweyer M, Theisen R, Wernig A (1997) Functional improvement of damaged adult mouse muscle by implantation of primary myoblasts. J Physiol 500:775–785

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Juretic N, Urzúa Munroe DJ, Jaimovich E, Riveros N (2007) Differential gene expression in skeletal muscle cells after membrane depolarization. J Cell Physiol 210:819–830

    Article  CAS  PubMed  Google Scholar 

  • Kern H, Salmons S, Mayr W, Rossini K, Carraro U (2005) Recovery of long-term denervated human muscles induced by electrical stimulation. Muscle Nerve 31:98–101

    Article  PubMed  Google Scholar 

  • Kubis HP, Hanke N, Scheibe RJ, Meissner JD, Gros G et al (2003) Ca2+ transients activate calcineurin/NFATc1 and initiate fast-to-slow transformation in a primary skeletal muscle culture. Am J Physiol Cell Physiol 285:C56–C63

    Article  CAS  PubMed  Google Scholar 

  • Langelaan ML, Boonen KJ, Rosaria-Chak KY, van der Schaft DW, Post MJ, Baaijens FPJ (2011) Advanced maturation by electrical stimulation: differences in response between C2C12 and primary muscle progenitor cells. Tissue Eng Regen Med 5:529–539

  • Manabe Y, Miyatake S, Takagi M, Nakamura M, Okeda A, Nakano T, Hirshman MF, Goodyear LJ, Fujii NL (2012) Characterization of an acute muscle contraction model using cultured C2C12 myotubes. PLoS One 7(12):e52592

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Martínez L, Pérez T, Mirasso CR, Manjarrez E (2007) Stochastic resonance in the motor system: effects of noise on the monosynaptic reflex pathway of the cat spinal cord. J Neurophysiol 97:4007–4016

  • McDonnell MD, Abbott D (2009) What is stochastic resonance? Definitions, misconceptions, debates, and its relevance to biology. PLoS Comput Biol 5:e1000348

    Article  PubMed Central  PubMed  Google Scholar 

  • Naaman SC, Stein RB, Thomas C (2000) Minimizing discomfort with surface neuromuscular stimulation. Neurorehabil Neural Repair 14:223–228

    CAS  PubMed  Google Scholar 

  • Nikolić N, Bakke SS, Kase ET, Rudberg I, Halle IF, Rustan AC et al (2012) Electrical pulse stimulation of cultured human skeletal muscle cells as an in vitro model of exercise. PLoSOne 7:e33203

    Article  Google Scholar 

  • Noto Y, Misawa S, Kanai K, Sato Y, Shibuya K, Isose S et al (2011) Activity-dependent changes in impulse conduction of single human motor axons: a stimulated single fiber electromyography study. Clin Neurophysiol 122:2512–2517

    Article  PubMed  Google Scholar 

  • Olson EN, Williams RS (2000) Remodeling muscles with calcineurin. BioEssays 22:510–519

    Article  CAS  PubMed  Google Scholar 

  • O’Reilly C, Pette D, Ohlendieck K (2003) Increased expression of the nicotinic acetylcholine receptor in stimulated muscle. Biochem Biophys Res Commun 300:585–591

  • Pedrotty DM, Koh J, Davis BH, Taylor DA, Wolf P, Niklason LE (2005) Engineering skeletal myoblasts: roles of three-dimensional culture and electrical stimulation. Am J Physiol Heart CircPhysiol 288:H1620–1626

    Article  CAS  Google Scholar 

  • Prior C, Dempster J, Marshall IG (1993) Electrophysiological analysis of transmission at the skeletal neuromuscular junction. J Pharmacol Toxicol Methods 30:1–17

    Article  CAS  PubMed  Google Scholar 

  • Robinson LR, Nielsen VK (1990) Limits of normal nerve function during high-frequency stimulation. Muscle Nerve 13:279–285

    Article  CAS  PubMed  Google Scholar 

  • Sciancalepore M, Afzalov R, Buzzin V, Jurdana M, Lorenzon P, Ruzzier F (2005) Intrinsic ionic conductances mediate the spontaneous electrical activity of cultured mouse myotubes. Biochim Biophys Acta 1720:117–124

    Article  CAS  PubMed  Google Scholar 

  • Sciancalepore M, Luin E, Parato G, Ren E, Giniatullin R, Fabbretti E et al (2012) Reactive oxygen species contribute to the promotion of the ATP-mediated proliferation of mouse skeletal myoblasts. Free RadicBiol Med 53:1392–1398

  • Stein RB, Chong SL, James KB, Kido A, Bell GJ, Tubman LA et al (2002) Electrical stimulation for therapy and mobility after spinal cord injury. Prog Brain Res 137:27–34

    Article  PubMed  Google Scholar 

  • Taccola G (2011) The locomotor central pattern generator of the rat spinal cord in vitro is optimally activated by noisy dorsal root waveforms. J Neurophysiol 106:872–884

    Article  PubMed  Google Scholar 

  • Thrasher A, Geoffrey MG, Popovic MR (2005) Reducing muscle fatigue due to functional electrical stimulation using random modulation of stimulation parameters. Artif Organs 29:453–458

    Article  PubMed  Google Scholar 

  • Valdes JA, Gaggero E, Hidalgo J, Leal N, Jaimovich E, Carrasco MA (2008) NFAT activation by membrane potential follows a calcium pathway distinct from other activity-related transcription factors in skeletal muscle cells. Am J Physiol Cell Physiol 294:C715–C725

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Zhang Y, Wang W, Cao Y, Han JS (2005) Effects of synchronous or asynchronous electroacupuncture stimulation with low versus high frequency on spinal opioid release and tail flick nociception. Exp Neurol 192:156–162

    Article  CAS  PubMed  Google Scholar 

  • Watson BV, Brown WF, Doherty TJ (2006) Frequency-dependent conduction block in carpal tunnel syndrome. Muscle Nerve 33:619–626

    Article  PubMed  Google Scholar 

  • Wehrle U, Dusterhoft S, Pette D (1994) Effects of chronic electrical stimulation on myosin heavy chain expression in satellite cell cultures derived from rat muscles of different fiber-type composition. Differentiation 58:37–46

    Article  CAS  PubMed  Google Scholar 

  • Zeng S, Holmes WR (2010) The effect of noise on CaMKII activation in a dendritic spine during LTP induction. J Neurophysiol 103:1798–1808

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Ziegler MD, Zhong H, Roy RR, Edgerton VR (2010) Why variability facilitates spinal learning. J Neurosci 30:10720–10726

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by Fondazione Kathleen Foreman Casali, Trieste, FRA2013-University of Trieste and Beneficentia Stiftung-Vaduz (Lichtenstein). The authors are particularly grateful to Dr. Andrew Constanti (UCL School of Pharmacy, UK) for critically reading the manuscript. We are also grateful to the physical therapists Rachele Menosso, Chiara Pinzini and Giuliana De Maio for the technical assistance during EMG acquisition.

Author information

Authors and Affiliations

  1. Department of Life Sciences & BRAIN Center, University of Trieste, Via Fleming 22, 34127, Trieste, Italy

    M. Sciancalepore, P. Lorenzon & G. Ziraldo

  2. Department of Neuroscience, SISSA, Trieste, Italy

    T. Coslovich & G. Taccola

Authors
  1. M. Sciancalepore
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Coslovich
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Lorenzon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Ziraldo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G. Taccola
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Sciancalepore.

Electronic supplementary material

Below is the link to the electronic supplementary material.

10974_2015_9424_MOESM1_ESM.tif

Supplementary Figure: At the end of the experiment, the stimulating electrode was moved far from the cells, although remaining in the recording bath (a). In this case, only stimulus artifact interference was recorded from the myotube (b). Band-pass filtering in b1 revealed the suppression of the interference. In c, a representative recording of spontaneous electrical activity elicited by a single myotube is shown and filtered with a band-pass filtering at 0.1–5 Hz in c1. It is clear that the information relative to the action potential frequency is not lost. Supplementary material 1 (TIFF 775 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sciancalepore, M., Coslovich, T., Lorenzon, P. et al. Extracellular stimulation with human “noisy” electromyographic patterns facilitates myotube activity. J Muscle Res Cell Motil 36, 349–357 (2015). https://doi.org/10.1007/s10974-015-9424-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10974-015-9424-2

Keywords

Search

Navigation