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Objective: Cell shrinkage has been documented as the earliest marker of apoptosis in all types of cells studied so far. Several research teams proposed that apoptotic volume decrease (AVD) contributes to triggering and/or development of the cell death machinery. This study examines the role of AVD in apoptosis of serum-deprived vascular smooth muscle cells (VSMC).

Design and Methods: VSMC were obtained from rat aorta and used between 10 and 16 passages. Cells transfected with a functional c-Myc analogue E1A-adenoviral protein (VSMC-E1A) were selected in the presence of 500 ug/ml geneticin. Apoptosis was estimated by phase-contrast microscopy, chromatin cleavage, DNA degradation, DAPI staining and caspase-3 activity. Cell volume was measured as [14C]-urea available space.

Results: Elevation of medium osmolality by addition of 300 mM mannitol led to reduction of VSMC volume by∼ 35 …