Calcium influx affects intracellular transport and membrane repair following nanosecond pulsed electric field exposure
| Autor: | Marjorie A. Kuipers, Caleb C. Roth, Gary L. Thompson, Danielle R. Dalzell, Bennett L. Ibey |
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| Rok vydání: | 2014 |
| Předmět: |
Biomedical Engineering
chemistry.chemical_element Nanotechnology Pyridinium Compounds CHO Cells Calcium Microtubules Calcium in biology law.invention Biomaterials chemistry.chemical_compound Cricetulus Electricity Confocal microscopy law Cricetinae Animals Propidium iodide Cytoskeleton Fluorescent Dyes Calcium metabolism Microscopy Confocal LAMP1 Chemistry Cell Membrane Biological Transport Atomic and Molecular Physics and Optics Electronic Optical and Magnetic Materials Quaternary Ammonium Compounds Membrane Biophysics Lysosomes Porosity |
| Zdroj: | Journal of biomedical optics. 19(5) |
| ISSN: | 1560-2281 |
| Popis: | The cellular response to subtle membrane damage following exposure to nanosecond pulsed electric fields (nsPEF) is not well understood. Recent work has shown that when cells are exposed to nsPEF, ion permeable nanopores (2 nm) are created in the plasma membrane in contrast to larger diameter pores (2 nm) created by longer micro- and millisecond duration pulses. Nanoporation of the plasma membrane by nsPEF has been shown to cause a transient increase in intracellular calcium concentration within milliseconds after exposure. Our research objective is to determine the impact of nsPEF on calcium-dependent structural and repair systems in mammalian cells. Chinese hamster ovary (CHO-K1) cells were exposed in the presence and absence of calcium ions in the outside buffer to either 1 or 20, 600-ns duration electrical pulses at 16.2 kV/cm, and pore size was determined using propidium iodide and calcium green. Membrane organization was observed with morphological changes and increases in FM1-43 fluorescence. Migration of lysosomes, implicated in membrane repair, was followed using confocal microscopy of red fluorescent protein-tagged LAMP1. Microtubule structure was imaged using mEmerald-tubulin. We found that at high 600-ns PEF dosage, calcium-induced membrane restructuring and microtubule depolymerization coincide with interruption of membrane repair via lysosomal exocytosis. |
| Databáze: | OpenAIRE |
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