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Role of chloride ions on the cell death caused by the increase of the plasma membrane permeability
Author(s) -
Hazama Akihiro,
Miyake Masao,
Saito Akiko,
Suzuki Daiji,
Wada Ikuo
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1463-b
Subject(s) - dids , membrane , lysosome , chemistry , biophysics , nucleus , membrane permeability , programmed cell death , cell membrane , staining , fluorescence , hela , fluorescein , permeability (electromagnetism) , cell , microbiology and biotechnology , biochemistry , apoptosis , biology , enzyme , physics , quantum mechanics , genetics
It is well known that the increase of the plasma membrane permeability causes the cell death. Detail mechanisms of this type of cell death are still unclear. We use amphotericin B (amB) for the membrane pore formation and investigate the ionic dependency on the cell death caused by amB. First we apply propidium iodede (PI) to the HeLa cells together with 10 μg/ml amB and observe the staining nucleus by PI using the fluorescent microscope. 2 hours after amB application, we could observe the PI signal in the nucleus, indicating that the large pores were formed after amB application. Cl − replacement by gluconate or Cl − channel blocker DIDS (0.5mM) inhibited the staining of nucleus after amB application. Next we stained the lysosome by FL‐labeled pepstatin A. We could observed fluorescent dots around the nucleus, which gradually disappeared after amB application. Low Cl − conditions or DIDS application inhibited the disappearance of the fluorescent dots by amB. These results suggest that the Cl − ions can enter into the cell after amB application and those ions may cause the disruption of lysosome, which enhances the membrane permeability increase by the attacking membrane proteins by the lysosomal enzymes