Effects of exogenous linoleic acid on fatty acid composition, receptor-mediated cAMP formation, and transport functions in rat astrocytes in primary culture.

Abstract

We have examined the effects of culturing neonatal rat-brain astrocytes in medium containing delipidated serum, with or without added linoleic acid (LA, 18:2 omega 6), on membrane fatty-acid composition and functions. After 18-21 days in culture, polyunsaturated fatty acids (PUFA) constituted approximately equal to 24 mol% of the total fatty acids in the astrocytes grown in delipidated media ("controls'); these proportions were increased by 35-40% to approximately equal to 33 mol% when the cells were supplemented with 35 microM LA. Notable differences in the PUFA profiles of the cells cultured with or without added LA included: (a) higher proportions of omega 6 PUFA in the LA-supplemented astrocytes (approximately equal to 25%, relative to approximately equal to 10% in controls) that were accompanied by an increase in the ratio of omega 6/omega 3 PUFA (from < 2 in controls to approximately equal to 5), and (b) higher proportions of 20:3 omega 9 and 22:3 omega 9 in the control astrocytes (> 5%) relative to the LA-supplemented cells (approximately equal to 1%). The major metabolites in the omega 6 PUFA-enriched cells were arachidonic (20:4 omega 6), adrenic (22:4 omega 6) and docosapentaenoic (22:5 omega 6) acids (15, 5 & 3 mol%, respectively). Enrichment of the astrocytes in omega 6 PUFA did not alter basal levels of cAMP, nor did it affect the amounts of cAMP formed in response to forskolin, isoproterenol, adenosine or histamine. However, dopamine-dependent increases in cAMP formation in the presence of the phosphodiesterase inhibitor, Ro 20-1724, were reduced by approximately equal to 25% relative to those in controls. LA supplementation modified uptake of [3H]adenosine into the astrocytes; values for Kt for a high affinity transport were increased relative to controls, and maximum capacity of a lower affinity process was reduced. Uptake of [3H]glutamate was not altered in the omega 6 PUFA-enriched astrocytes. This study demonstrated that cultured astrocytes take up exogenous linoleic acid and incorporate its metabolites into phospholipid, and that the resulting changes in membrane PUFA composition modify only specific cell functional properties.