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Location-dependent photogeneration of calcium waves in HeLa cells

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Abstract

The calcium ion (Ca2+) concentrations in a cell are responsible for the control of vital cellular functions and have been widely studied as a means to investigate and control cell activities. Here, we demonstrate Ca2+ wave generation in HeLa cells by femtosecond laser irradiation and show unexpected properties of the Ca2+ release and propagation. When the laser was focused in the cell cytoplasm, Ca2+ release was independent of both external Ca2+ influx and the phosphoinositide-phospholipase C (PLC) signaling pathway. The nucleus was not a susceptible target for laser-induced Ca2+ release, whereas irradiation of the plasma membrane produced evidence of transient poration, through which the extracellular solution could enter the cell. By chelating extracellular Ca2+, we found that laser-induced influx of ethylene glycol tetra-acetic acid (EGTA) can compete with calcium-induced calcium release and significantly delay or suppress the onset of the Ca2+ wave in the target cell. Intercellular Ca2+ propagation was adenosine triphosphate-dependent and could be observed even when the target cell cytosolic Ca2+ rise was suppressed by influx of EGTA. The irradiation effect on overall cell viability was also tested and found to be low (85% at 6h after irradiation by 60 mW average power). Laser-induced Ca2+ waves can be reliably generated by controlling the exposure and focal position and do not require the presence of caged Ca2+. The technique has the potential to replace other methods of Ca2+ stimulation, which either require additional caged molecules in the cell or do not have an interaction that is as well localized.

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Authors and Affiliations

  1. Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, 565-0871, Suita, Osaka, Japan

    Shigeki Iwanaga, Tomoyuki Kaneko, Katsumasa Fujita, Osamu Nakamura & Satoshi Kawata

  2. Department of Frontier Biosciences, Graduate School of Frontier Biosciences, Osaka University, 2-1 Yamadaoka, 565-0871, Suita, Osaka, Japan

    Nicholas Smith & Osamu Nakamura

  3. Department of Pathology and Cell Regulation, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamikyo-Ku, 602-0841, Kyoto, Japan

    Tetsuro Takamatsu

  4. RIKEN, 2-1 Hirosawa, 351-0198, Wako, Saitama, Japan

    Satoshi Kawata

Authors
  1. Shigeki Iwanaga
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  2. Tomoyuki Kaneko
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  3. Katsumasa Fujita
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  4. Nicholas Smith
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  5. Osamu Nakamura
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  6. Tetsuro Takamatsu
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  7. Satoshi Kawata
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Correspondence to Nicholas Smith.

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Iwanaga, S., Kaneko, T., Fujita, K. et al. Location-dependent photogeneration of calcium waves in HeLa cells. Cell Biochem Biophys 45, 167–176 (2006). https://doi.org/10.1385/CBB:45:2:167

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  • DOI: https://doi.org/10.1385/CBB:45:2:167

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