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The Journal of Lipid Research, Vol. 40, 495-503, March 1999
Copyright © 1999 by Lipid Research, Inc.

Original Article

Plasmalogen status influences docosahexaenoic acid levels in a macrophage cell line: insights using ether lipid-deficient variants

Correspondence to: Raphael A. Zoeller

Previously, this laboratory reported the isolation of variants, RAW.12 and RAW.108, from the macrophage-like cell line RAW 264.7 that are defective in plasmalogen biosynthesis [Zoeller, R.A. et al. 1992. J. Biol. Chem. 267: 8299–8306]. Fatty acid analysis showed significant changes in the mutants in the ethanolamine phospholipids (PE), the only phospholipid class in which the plasmalogen species, plasmenylethanolamine, contributes significantly. Within the PE fraction, docosapentaenoic (DPA; 22:5n–3) and docosahexaenoic (DHA; 22:6n–3) acids were reduced by approximately 50% in the variants while the levels of arachidonic acid (AA; 20:4n–6) remained unaffected. The decrease in DHA was accompanied by a 50% decrease in labeling PE with [³H]DHA over a 90-min period. Restoration of plasmenylethanolamine by supplementing the growth medium with sn -1-hexadecylglycerol (HG) completely reversed these changes in RAW.108. Pre-existing pools of plasmenylethanolamine were not required for restoration of normal [³H]DHA labeling; addition of HG only during the labeling period was sufficient. Due to the loss of 1'-desaturase in RAW.12, HG supplementation resulted in the accumulation of plasmenylethanolamine's immediate biosynthetic precursor, plasmanylethanolamine. Even though this latter phospholipid contained only the ether functionality (lacking the vinyl ether double bond) it was sufficient to restore wild type-like fatty acid composition and DHA labeling of the ethanolamine phospholipids, identifying the ether bond as a structural determinant for this specificity.

In summary, we have used these mutants to establish that the plasmalogen status of a cell can influence the levels of certain polyunsaturated fatty acids. These results support the notion that certain polyunsaturated fatty acids, such as DHA, can be selectively targeted to plasmalogens and that this targeting occurs during de novo biosynthesis, or shortly thereafter, through modification of nascent plasmalogen pools.—Gaposchkin, D. P., and R. A. Zoeller. Plasmalogen status influences docosahexaenoic acid levels in a macrophage cell line: insights using ether lipid-deficient variants. J. Lipid Res. 1999. 40: 495–503.

Supplementary key words: arachidonic acid, docosahexaenoic acid, ether lipid, plasmalogen, sn -1-hexadecylglycerol

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