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Originally published In Press as doi:10.1074/jbc.M004446200 on July 19, 2000

J. Biol. Chem., Vol. 275, Issue 45, 35215-35223, November 10, 2000
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Inhibition of Neuronal Apoptosis by Docosahexaenoic Acid (22:6n-3)
ROLE OF PHOSPHATIDYLSERINE IN ANTIAPOPTOTIC EFFECT*

Hee-Yong KimDagger , Mohammed Akbar, Audrey Lau, and Lisa Edsall

From the Section of Mass Spectrometry, Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, Maryland 20852

Enrichment of Neuro 2A cells with docosahexaenoic acid (22:6n-3) decreased apoptotic cell death induced by serum starvation as evidenced by the reduced DNA fragmentation and caspase-3 activity. The protective effect of 22:6n-3 became evident only after at least 24 h of enrichment before serum starvation and was potentiated as a function of the enrichment period. During enrichment 22:6n-3 incorporated into phosphatidylserine (PS) steadily, resulting in a significant increase in the total PS content. Similar treatment with oleic acid (18:1n-9) neither altered PS content nor resulted in protective effect. Hindering PS accumulation by enriching cells in a serine-free medium diminished the protective effect of 22:6n-3. Membrane translocation of Raf-1 was significantly enhanced by 22:6n-3 enrichment in Neuro 2A cells. Consistently, in vitro biomolecular interaction between PS/phosphatidylethanolamine /phosphatidylcholine liposomes, and Raf-1 increased in a PS concentration-dependent manner. Collectively, enrichment of neuronal cells with 22:6n-3 increases the PS content and Raf-1 translocation, down-regulates caspase-3 activity, and prevents apoptotic cell death. Both the antiapoptotic effect of 22:6n-3 and Raf-1 translocation are sensitive to 22:6n-3 enrichment-induced PS accumulation, strongly suggesting that the protective effect of 22:6n-3 may be mediated at least in part through the promoted accumulation of PS in neuronal membranes.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Section of Mass Spectrometry, LMBB/NIAAA, NIH, 12420 Parklawn Dr., Rockville, MD 20852. Tel.: 301-402-8746; Fax: 301-594-0035; E-mail: hykim@nih.gov.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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