Evaluating muscular calcium dynamics upon pulsed electric field exposure

James Mancillas; Reinhardt Knerr; Anna V. Sedelnikova; Bryan Gamboa; Mara D. Casebeer; Ronald Barnes Jr.; Gleb Tolstykh; Bennett Ibey; Christopher M. Valdez

doi:10.1117/12.2553090

11 March 2020 Evaluating muscular calcium dynamics upon pulsed electric field exposure

James Mancillas, Reinhardt Knerr, Anna V. Sedelnikova, Bryan Gamboa, Mara D. Casebeer, Ronald Barnes Jr., Gleb Tolstykh, Bennett Ibey, Christopher M. Valdez

Author Affiliations +

Proceedings Volume 11238, Optical Interactions with Tissue and Cells XXXI; 112380Y (2020) https://doi.org/10.1117/12.2553090
Event: SPIE BiOS, 2020, San Francisco, California, United States

Abstract

Nanosecond pulsed electric fields (nsPEF) are high voltage (1-15 kV/cm) nanosecond energy waveforms that can impact cellular activity. On a physical level, nsPEF generates transient membrane perturbations in the form of nanopores to allow cation influx resulting in localized membrane depolarization. On a physiological level, nsPEF exposure can activate second messenger cascades resulting in subcellular modulation that lasts beyond the nsPEF duration. An ongoing challenge is to characterize the physiological events induced by nsPEF exposure, and potential to interplay with physical effects induced by the pulse. In our laboratory, C2C12 immortalized mouse myoblast cells have been demonstrated to be a useful in vitro model, by differentiating these progenitors into terminally transformed myotubes. We are not only able to further investigate the fundamental subcellular mechanisms activated by pulsed electric fields, but monitor muscle contraction as a functional output. From our previous efforts, we quantified calcium-green uptake as a measurement of cellular calcium uptake across a sweep of applied pulsed electric field voltages. To extend on these findings, we evaluated calcium dynamics in the intracellular space of myotubes. Given that sarcoplasmic reticulum efflux is required for muscle contraction, we tested the physiological role of the ryanodine receptor during pulsed electric field exposure on myotubes. By blocking the Ryanodine receptor with a competitive antagonist, we reduced nsPEF -induced calcium dynamics activation by 58.36% in media with calcium. Our results are the first to demonstrate that the Ryanodine receptor complex is a subcellular candidate responsible for generating calcium responses upon nsPEF exposure in myotubes.

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James Mancillas, Reinhardt Knerr, Anna V. Sedelnikova, Bryan Gamboa, Mara D. Casebeer, Ronald Barnes Jr., Gleb Tolstykh, Bennett Ibey, and Christopher M. Valdez "Evaluating muscular calcium dynamics upon pulsed electric field exposure", Proc. SPIE 11238, Optical Interactions with Tissue and Cells XXXI, 112380Y (11 March 2020); https://doi.org/10.1117/12.2553090

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