Docosahexaenoic acid (DHA) alters the structure and composition of membranous vesicles exfoliated from the surface of a murine leukemia cell line

AbstractMembrane lipid microdomains are regions of the membrane thought to be functionally important, but which have remained poorly characterized because they have proven to be difficult to isolate. The exfoliation of small membranous vesicles from the cell surface is a continuous and normal activity in many cells. If microdomains are relatively large or stable, they may influence the structure and composition of exfoliated vesicles, which are easy to isolate. We tested the ability of docosahexaenoic acid (DHA), a fatty acid proposed to alter the structure of microdomains, to change the structure and composition of vesicles exfoliated from a murine leukemia cell line. Cells were cultured in normal and DHA-enriched media for 72 h, then washed and given a 15-h exfoliation period. Afterwards, the pooled vesicles and their parent plasma membrane were collected and analyzed. Vesicles and plasma membrane from cells grown in normal culture medium had similar fatty acid compositions, including equal, and low, proportions of DHA, but the vesicles had much more cholesterol and displayed higher anisotropy than the plasma membrane. When cells were grown in DHA-enriched medium, both the plasma membrane and exfoliated vesicles had 10-fold elevated levels of DHA in their phospholipids, with the DHA displacing other polyunsaturates. These cells released vesicles having significantly reduced levels of cholesterol and monoenoic fatty acids than those in normal culture. The anisotropy of these vesicles was also dramatically reduced. These data are consistent with DHA altering the structure and composition of membrane microdomains on the cell surface, and suggest that exfoliated vesicles may prove useful in the further study of membrane microdomains