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Titolo:
Changes in elemental content during apoptotic cell death studied by electron probe X-ray microanalysis
Autore:
Fernandez-Segura, E; Canizares, FJ; Cubero, MA; Warley, A; Campos, A;
Indirizzi:
Univ Granada, Sch Med, Dept Cell Biol, E-18071 Granada, Spain Univ Granada Granada Spain E-18071 ept Cell Biol, E-18071 Granada, Spain Univ Granada, Inst Neurosci, E-18071 Granada, Spain Univ Granada GranadaSpain E-18071 Inst Neurosci, E-18071 Granada, Spain St Thomas Hosp, Rayne Inst, Dept Ophthalmol, EM Unit, London SE1 7EH, England St Thomas Hosp London England SE1 7EH , EM Unit, London SE1 7EH, England
Titolo Testata:
EXPERIMENTAL CELL RESEARCH
fascicolo: 2, volume: 253, anno: 1999,
pagine: 454 - 462
SICI:
0014-4827(199912)253:2<454:CIECDA>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
ENDOGENOUS ENDONUCLEASE ACTIVATION; INDUCED THYMOCYTE APOPTOSIS; DNA FRAGMENTATION; BIOCHEMICAL PATHWAYS; SMOOTH-MUSCLE; T-LYMPHOCYTES; PROTEASE; K+; DEGRADATION; POTASSIUM;
Keywords:
apoptosis; electron probe X-ray microanalysis; cell shrinkage; sodium; potassium; chlorine;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
61
Recensione:
Indirizzi per estratti:
Indirizzo: Fernandez-Segura, E Univ Granada, Sch Med, Dept Cell Biol, E-18071 Granada, Spain Univ Granada Granada Spain E-18071 71 Granada, Spain
Citazione:
E. Fernandez-Segura et al., "Changes in elemental content during apoptotic cell death studied by electron probe X-ray microanalysis", EXP CELL RE, 253(2), 1999, pp. 454-462

Abstract

Recent data suggest that changes in ionic content, primarily potassium, play a pivotal role in the progression of apoptosis. However, the changes in total element content, i.e., sodium (Na), magnesium (Mg), phosphorous (P), chlorine (CI), potassium (K), and calcium (Ca), during apoptosis have not been evaluated. Electron probe X-ray microanalysis (EPXMA) was used to measure total element content in U937 cells before and after the induction of apoptosis. As an experimental model we used U937 cells irradiated with ultraviolet (UV) light. Apoptosis was evaluated with phase-contrast microscopy, with scanning and transmission electron microscopy, and with the fluorescentdye bis-benzimide (Hoechst 33342).]Plasma membrane permeability as a measure of cell death was determined by trypan blue dye exclusion. To investigate element content with EPXMA, cells were cryoprepared, i.e., cryofixed and freeze-dried, and analyzed as whole cells using a scanning electron microscope. We found that the UV irradiation induced rapid (within 2 h) morphological changes associated with apoptosis, such as plasma membrane blebbing, condensation of the chromatin, and the formation of membrane-bound apoptotic bodies. At this time, 95% of the apoptotic cells excluded trypan blue dye. EPXMA results demonstrated that UV light-irradiated apoptotic cells (cells with membrane-bound apoptotic bodies) had a lower Cl content (P < 0.001) and K content (P < 0.001) and, a higher Na content (P < 0.001) in comparison with nonirradiated control cells. Also, P and Ca content was higher in apoptotic cells than in control cells, but this difference did not reach statistical significance. No differences were found in Mg. These data indicated that morphological changes characteristic of apoptotic cell death are related with significant changes in sodium, chlorine, and potassium content. In addition, we demonstrated that these changes in elemental composition were not associated with loss of cell membrane integrity. (C) 1999 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/09/20 alle ore 19:28:01