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Proceedings Paper

Cytosolic acidification and mitochondrial dysfunction during apoptosis as probed by dual emission probes and microspectrofluorometry
Author(s): Rajae Belhoussine; Valerie Palissot; Hamid Morjani; Jean Dufer; Michel Manfait
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Paper Abstract

Cytosolic acidification and mitochondrial dysfunction have been reported to be concomitant with apoptosis. In our study, we report on the use of carboxy-SNARF-1-AM and JC1 probes in association with confocal scanning microspectrofluorometry to examine respectively cytosolic pH and mitochondrial membrane potential during camptothecin (CPT) and daunorubicin (DNR) induced apoptosis in human leukemic HL60 cells. Emission intensity ratios (I580 nm/I640 nm) of carboxy-SNARF-1 spectra lead to determine cytosolic pH. Whereas, JC1 was able to form J-aggregates respectively from green (530 nm) to yellow-orange (590 nm) as mitochondrial membrane becomes more polarized. Our results show that control cells presented a neutral cytosolic pH (7.26 plus or minus 0.08). The cells induced in apoptosis in presence of 1 (mu) M CPT or DNR during 6hrs, undergo significant acidification of the cytosol (7.00 plus or minus 0.06 and 7.02 plus or minus 0.05 respectively). Decrease of I590 nm/I530 nm emission ratio of JC1 spectra (64 plus or minus 5% compared to control) was observed only with CPT. We conclude that cytosolic acidification and mitochondrial dysfunction are early events in apoptosis and may be essential for genome destruction. Confocal scanning microspectrofluorometry and dual ratiometric fluorescent probes appear to be a powerful approach for the study of ion response in living cells.

Paper Details

Date Published: 29 April 1998
PDF: 4 pages
Proc. SPIE 3260, Optical Investigations of Cells In Vitro and In Vivo, (29 April 1998); doi: 10.1117/12.307086
Show Author Affiliations
Rajae Belhoussine, Faculte de Pharmacie/Univ. de Reims Champagne-Ardenne (France)
Valerie Palissot, Faculte de Pharmacie/Univ. de Reims Champagne-Ardenne (France)
Hamid Morjani, Faculte de Pharmacie/Univ. de Reims Champagne-Ardenne (France)
Jean Dufer, Faculte de Pharmacie/Univ. de Reims Champagne-Ardenne (France)
Michel Manfait, Faculte de Pharmacie/Univ. de Reims Champagne-Ardenne (France)


Published in SPIE Proceedings Vol. 3260:
Optical Investigations of Cells In Vitro and In Vivo
Robert C. Leif; Daniel L. Farkas; Robert C. Leif; Bruce J. Tromberg, Editor(s)

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