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Department of Paediatrics, University of British Columbia, Vancouver, BC, Canada, V5Z 4H4
1 To whom correspondence should be addressed. e-mail: sinnis{at}nutrition.ubc.ca
The phospholipids, particularly phosphatidylethanolamine, of brain gray matter are enriched with docosahexaenoic acid (22:6n-3). The importance of uptake of preformed 22:6n-3 from plasma compared with synthesis from the 
-linolenic acid (18:3n-3) precursor in brain is not known. Deficiency of 18:3n-3 results in a compensatory increase in the n-6 docosapentaenoic acid (22:5n-6) in brain, which could be formed from the precursor linoleic acid (18:2n-6) in liver or brain. We studied n-3 and n-6 fatty acid incorporation in brain astrocytes cultured in chemically defined medium using delipidated serum supplemented with specific fatty acids. High performance liquid chromatography with evaporative light scattering detection and gas liquid chromatography were used to separate and quantify cell and media lipids and fatty acids. Although astrocytes are able to form 22:6n-3, incubation with 18:3n-3 or eicosapentaenoic acid (20:5n-3) resulted in a time and concentration dependent accumulation of 22:5n-3 and decrease in 22:6n-3 g/g cell fatty acids. Astrocytes cultured with 18:2n-6 failed to accumulate 22:5n-6.
Astrocytes secreted cholesterol esters (CE) and phosphatidylethanolamine containing saturated and monounsaturated fatty acids, and arachidonic acid (20:4n-6) and 22:6n-3. These studies suggest conversion of 22:5n-3 limits 22:6n-3 synthesis, and show astrocytes release fatty acids in CE.
Abbreviations: CE, cholesterol ester; dbc-AMP, dibutyryl cyclic AMP; ELSD, evaporative light scattering detection; FCS, fetal calf serum; GFAP, glial fibrillary acidic protein; GLC, gas liquid chromatography; HPLC, high performance liquid chromatography; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; TG, triglyceride
Supplementary key words astrocyte • docosahexaenoic acid • docosapentaenoic acid • alpha linolenic acid • desaturation • elongation • cholesterol ester • phosphatidylethanolamine
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