1H NMR-visible mobile lipid domains correlate with cytoplasmic lipid bodies in apoptotic T-lymphoblastoid cells.
The presence of nuclear magnetic resonance (NMR)-visible mobile lipid (ML) domains in apoptotic lymphoblasts suggests alterations in neutral lipid metabolism and compartmentation during programmed cell death. The detection of similar ML signals in activated lymphocytes raises questions about common mechanisms of ML formation during apoptosis and upon lymphoblast stimulation. Structure and subcellular localization of ML domains were therefore investigated by NMR, fluorescence and electron microscopy in Jurkat T-lymphoblasts either induced to apoptosis (by anthracyclines or dexamethasone or by serum deprivation) or activated by phorbol myristate acetate (PMA) plus ionomycin. ML contents in drug-treated cells correlated linearly with apoptosis, irrespective of the specific inducer and cell cycle arrest phase (r = 0.993, P < 0.001). Similar ML levels were measured in drug-induced apoptotic cells (A approximately 30-40%) and in non-apoptotic PMA/ionomycin-treated lymphoblasts (72 h). Lower ML contents were instead formed in serum-deprived apoptotic cells, with respect to controls. Increases in ML signals were associated, in either apoptotic or activated cells, with the accumulation of cytoplasmic, osmophilic lipid bodies (diameter < or = 1.0 microm), surrounded by own membrane, possessing intramembrane particles. The results support the hypothesis that ML are formed in the cytoplasm of drug-induced apoptotic cells during an early, 'biochemically active' phase of programmed cell death.
Di Vito
M
M
Laboratory of Cell Biology, Istituto Superiore di Sanità, Rome, Italy.
Lenti
L
L
Knijn
A
A
Iorio
E
E
D'Agostino
F
F
Molinari
A
A
Calcabrini
A
A
Stringaro
A
A
Meschini
S
S
Arancia
G
G
Bozzi
A
A
Strom
R
R
Podo
F
F
D017209:Apoptosis; D003593:Cytoplasm / Q000378:metabolism*; D005456:Fluorescent Dyes; D005614:Freeze Fracturing; D006801:Humans; D015759:Ionomycin; D019169:Jurkat Cells; D008055:Lipids / Q000032:analysis*; D008213:Lymphocyte Activation; D009682:Magnetic Resonance Spectroscopy; D008854:Microscopy, Electron; D010078:Oxazines; D013601:T-Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D013755:Tetradecanoylphorbol Acetate
D005456:Fluorescent Dyes; D008055:Lipids; D010078:Oxazines; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate; C044808:nile red