Variations Of Intracellular Ca2+ Mobilization Initiated By Nanosecond And Microsecond Electrical Pulses In Hela Cells

Goal: Herein, the variations in transient Ca2+ mobilizations in HeLa cells exposed to a single, non-thermal pulsed electric field (PEF) are described. Methods: Three PEF waveforms categorized by pulse duration and intensity were used to deduce the kinetics involved in Ca2+ mobilization. A fast microscopic fluorescent imaging system and a fluorescent molecular probe were used to observe transient intracellular Ca2+ mobilization after pulse exposure. The sources and pathways in the transient Ca2+ mobilizations were investigated using an inhibitor of inositol-1,4,5-trisphosphate receptor (IP3 R) on the endoplasmic reticulum (ER) along with a Ca2+-free buffer. Results: When exposed to the 10-mu s-long PEF, the Ca2+ concentration increased mainly at the cathodic region near the membrane. However, Ca2+ concentration increased at both anodic and cathodic regions when Na+ concentration in the buffer was reduced. Ca2+ concentration increased only in the presence of extracellular Ca2+. Conclusion: These results suggest that the 10-mu s PEF takes a large amount of extracellular Na+ into the cell through the electropermeabilized plasma membrane, especially at the anodic side, resulting in the suppression of the Ca2+ influx. On the contrary, the 20-ns-long PEF increased Ca2+ concentration in the surrounding region of the nucleus only in the presence of extracellular Ca2+. The PEF exposure with inhibition of the IP3R indicates that increased Ca2+ ions are released from the ER via the activated IP3 R. Significance: These mechanisms could induce specific cell responses, such as Ca2+ oscillations, Ca2+ waves, and Ca2+ puffs.