Plasma membrane phospholipid asymmetry precedes DNA fragmentation in different apoptotic cell models.
Biochemical alterations occurring in many cell types during apoptosis include the loss of plasma membrane phospholipid asymmetry and nuclear DNA fragmentation. Annexin V staining detects phosphatidylserine translocation into the outer plasma membrane layer occurring during cell death, while the in situ tailing (IST or TUNEL) reaction labels the DNA strand breaks typical of apoptosis. To compare the time course of these processes we investigated methylprednisolone-induced apoptosis of rat thymocytes, topoisomerase inhibitor-induced apoptosis in the human histiocytic lymphoma cell line U937, and serum deprivation-induced apoptosis in the rat pheochromocytoma cell line, PC12. At all time points, FACS analysis and quantitative fluorescence light microscopy showed a higher proportion of annexin V-positive than IST-positive cells, with significantly different time courses in the apoptotic cell models investigated (Anova test). Results were confirmed by confocal microscopy. Our data indicate that the exposure of phosphatidylserine, a potential phagocyte recognition signal on the cell surface of apoptotic cells in vivo, precedes DNA strand breaks during apoptosis in different cell types.
Chan
A
A
Department of Neurology, Clinical Research Group for Multiple Sclerosis, Julius-Maximilians-University, Würzburg, Germany.
Reiter
R
R
Wiese
S
S
Fertig
G
G
Gold
R
R
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D002166:Camptothecin / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects; D053938:DNA Fragmentation; D004273:DNA, Neoplasm / Q000378:metabolism; D005434:Flow Cytometry; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D008563:Membrane Lipids / Q000378:metabolism*; D008775:Methylprednisolone / Q000494:pharmacology; D018613:Microscopy, Confocal; D016716:PC12 Cells / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew; D013950:Thymus Gland / Q000166:cytology; D020298:U937 Cells / Q000378:metabolism
D017304:Annexin A5; D004273:DNA, Neoplasm; D008563:Membrane Lipids; D010718:Phosphatidylserines; D008775:Methylprednisolone; D002166:Camptothecin