Mouse blastocysts release a lipid which activates anandamide hydrolase in intact uterus.
Anandamide (N-arachidonoylethanolamine, AEA) is a major endocannabinoid, known to impair mouse pregnancy and embryo development and to induce apoptosis in blastocysts. Here we show that mouse blastocysts rapidly (within 30 min of culture) release a soluble compound, that increases by approximately 2.5-fold the activity of AEA hydrolase (fatty acid amide hydrolase, FAAH) present in the mouse uterus, without affecting FAAH gene expression at the translational level. This "FAAH activator" was produced by both trophoblast and inner cell mass cells, and its initial biochemical characterization showed that it was fully neutralized by adding lipase to the blastocyst-conditioned medium (BCM), and was potentiated by adding trypsin to BCM. Other proteases, phospholipases A(2), C or D, DNAse I or RNAse A were ineffective. BCM did not affect the AEA-synthesizing phospholipase D, the AEA-binding cannabinoid receptors, or the selective AEA membrane transporter in mouse uterus. The FAAH activator was absent in uterine fluid from pregnant mice and could not be identified with any factor known to be released by blastocysts. In fact, platelet-activating factor inhibited non-competitively FAAH in mouse uterus extracts, but not in intact uterine horns, whereas leukotriene B(4) or prostaglandins E(2) and F(2)alpha had no effect. Overall, it can be suggested that blastocysts may protect themselves against the noxious effects of uterine endocannabinoids by locally releasing a lipid able to cross the cell membranes and to activate FAAH. The precise molecular identity of this activator, the first ever reported for FAAH, remains to be elucidated.
Department of Biomedical Sciences, University of Teramo, Piazza A. Moro 45, Teramo 64100, Italy. Maccarrone@vet.unite.it
M
M
Maccarrone
M
M
DeFelici
F G
FG
Klinger
N
N
Battista
F
F
Fezza
E
E
Dainese
G
G
Siracusa
A
A
Finazzi-Agrò
D000581:Amidohydrolases; D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D017077:Culture Media, Conditioned; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D005260:Female; D050356:Lipid Metabolism; D051379:Mice; D018345:Mice, Knockout; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D010739:Phospholipase D / Q000378:metabolism; D011247:Pregnancy; D043882:Receptors, Cannabinoid / Q000378:metabolism; D014599:Uterus / Q000378:metabolism*; D000099252:Fatty Acid Amide Hydrolases
D000581:Amidohydrolases; D017077:Culture Media, Conditioned; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D010739:Phospholipase D; D043882:Receptors, Cannabinoid; D000099252:Fatty Acid Amide Hydrolases