Article ID abbi.1999.1410, available online at http://www.idealibrary.com on
Biosynthesis and Inactivation of
-Docosahexaenoylethanolaminein Bovine Retina
I. Delton-Vandenbroucke,† A. Milone,*
M. Lagarde,† and V. Di Marzo*
Istituto per la Chimica di Molecole di Interesse Biologico, CNR, Via Toiano 6, 80072 Arco Felice, Napoli, Italy;and
Institut National de la Sante´ et de la Recherche Me´dicale, U352 Biochimie and Pharmacologie, INSA de Lyon, 20, avenue A. Einstein, 69621 Villeurbanne (Cedex), France
Received June 10, 1999, and in revised form July 26, 1999
-Arachidonoylethanolamine (anandamide; AEA)and 2-arachidonoylglycerol (2-AG), the two proposedendogenous agonists of cannabinoid receptors, andthe putative AEA biosynthetic precursor,
-arachido-noylphosphatidylethanolamine (NArPE), were identi-ﬁed in bovine retina by means of gas chromatography– electron impact mass spectrometry (GC-EIMS). Thistechnique also allowed us to identify
-docosahex-anoylethanolamine (DHEA) and 2-docosahexanoylg-lycerol (2-DHG), two derivatives of docosahexaenoicacid (DHA), one of the most abundant fatty acids es-teriﬁed in retina phospholipids and necessary for op-timal retinal function.
-Docosahexaenoylphosphati-dylethanolamine (NDHPE), the potential biosyntheticprecursor for DHEA, was also found. The fatty acidcomposition of the
-2 positions of bovineretina’s most abundant phospholipid classes, also de-termined here, were in agreement with a phospholip-id-dependent mechanism for 2-AG, 2-DHG, AEA, andDHEA biosynthesis, as very high levels of polyunsatu-rated fatty acids, including DHA, were found on the
-2 position of phosphatidylcholine (PC) and -etha-nolamine (PE), and measurable amounts of di-docosa-hexanoyl-PC and -PE, two potential biosynthetic pre-cursors of NDHPE, were detected. Accordingly, wefound that isolated particulate fractions from bovineretina could release AEA and DHEA in a time-depen-dent fashion. Finally, a fatty acid amide hydrolase(FAAH)-like activity with subcellular distribution andpH dependency similar to those reported for the brainenzyme was also detected in bovine retina. This activ-ity was inhibited by FAAH inhibitors, phenylmethyl-sulfonyl ﬂuoride and arachidonoyltriﬂuoromethyl-ketone, and appeared to recognize DHEA with a lowerefﬁciency than AEA. These data indicate that AEA andits congeners may play a physiological role in themammalian eye.© 1999 Academic Press
cannabinoids; anandamide; 2-arachido-noylglycerol; bovine retina; docosahexaenoic acid;eye.