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The Journal of Membrane Biology

, Volume 248, Issue 1, pp 117–123 | Cite as

Cell Death Parameters as Revealed by Whole-Cell Patch-Clamp and Interval Weighted Spectra Averaging: Changes in Membrane Properties and Current Frequency of Cultured Mouse Microglial Cells Induced by Glutaraldehyde

  • Aleksandar Kalauzi
  • Ljiljana Nikolić
  • Danijela Savić
  • Ksenija Radotić
Article

Abstract

The physiological and biochemical factors that lead to cell death have not been recognized completely. To our knowledge, there are no data on the bioelectric parameters that characterize early period of cell death, as well as on the appearance of related membrane current frequencies. We studied early parameters of glutaraldehyde (GA)-induced cell death, by examining the membrane properties of mouse microglia using the whole-cell patch-clamp technique. In addition, we investigated the GA-induced changes in the membrane current frequency, to see if characteristic frequencies would appear in dying cell. For data analysis, we applied a new approach, an improved multiple moving window length analysis and interval weighted spectra averaging (IWSA). We chose GA for its ability to induce almost instantaneous cell death. The 0.6 % GA did not induce changes in the bioelectric membrane properties of microglia. However, the 3 % GA caused significant decrease of membrane capacitance and resistance accompanied by the prominent increase in the membrane currents and nearly ohmic current response of microglial cells. These data indicate that 3 % GA caused complete loss of the membrane function consequently inducing instantaneous cell death. The membrane function loss was characterized by appearance of the 1.26–4.62 Hz frequency peak in the IWSA spectra, while no significant increase of amplitudes could be observed for cells treated with 0.6 % GA. To our knowledge, this is the first record of a frequency associated with complete loss of the membrane function and thus can be considered as an early indicator of cell death.

Keywords

Microglia Interval weighted spectra averaging Membrane current frequency Patch-clamp Whole-cell current 

Notes

Acknowledgments

This work was supported by the Grants III45012, III41014, OI 173022, from the Ministry of Education, Science and Technological Development of the Republic of Serbia. The authors are grateful to Sutter Instrument Company for donating the MP-285 Micromanipulator System, and Tecella Company for donating the Tecella Pico 2 amplifier.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Aleksandar Kalauzi
    • 1
  • Ljiljana Nikolić
    • 2
  • Danijela Savić
    • 2
  • Ksenija Radotić
    • 1
  1. 1.Institute for Multidisciplinary ResearchUniversity of BelgradeBelgradeSerbia
  2. 2.Institute for Biological Research “Siniša Stanković”University of BelgradeBelgradeSerbia

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