Regional distribution and age‐dependent expression of N‐acylphosphatidylethanolamine‐hydrolyzing phospholipase D in rat brain

• Published in: Journal of neurochemistry Vol. 94; no. 3; pp. 753 - 762
• Main Authors: Morishita, Jun, Okamoto, Yasuo, Tsuboi, Kazuhito, Ueno, Masaki, Sakamoto, Haruhiko, Maekawa, Nobuhiro, Ueda, Natsuo
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.08.2005; Blackwell; Blackwell Publishing Ltd
• Subjects: Age; Age Factors; Aging - physiology; anandamide; Animals; Animals, Newborn; Biochemistry and metabolism; Biological and medical sciences; Blotting, Western - methods; Brain; Brain - anatomy & histology; Brain - enzymology; Brain Chemistry - physiology; Central nervous system; Cercopithecus aethiops; Cetrimonium Compounds - metabolism; COS Cells; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; endocannabinoid; Enzymes; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation, Enzymologic - physiology; Male; Medical sciences; Neurology; N‐acylethanolamine; N‐acylphosphatidylethanolamine; N‐acylphosphatidylethanolamine‐hydrolyzing phospholipase D; p-Chloromercuribenzoic Acid - metabolism; Phosphatidylethanolamines - genetics; Phosphatidylethanolamines - metabolism; Phospholipase D - genetics; Phospholipase D - metabolism; rat brain; Rats; Recombinant Proteins - metabolism; Reverse Transcriptase Polymerase Chain Reaction - methods; Rodents; Subcellular Fractions - enzymology; Vertebrates: nervous system and sense organs; Heart; endocannabinoid; Rat; Central nervous system; Esterases; Phosphoric diester hydrolases; Encephalon; N-acylethanol- amine; Chlorides; Development; Immunoblotting assay; Thalamus; N-acylphosphatidylethanolamine; anandamide; Age; Phosphatidylethanolamine; Enzyme; Rodentia; rat brain; Phospholipase D; Protein; Vertebrata; Mammalia; Hydrolases; Circulatory system; N-acylphosphatidyl- ethanolamine-hydrolyzing phospholipase D
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/j.1471-4159.2005.03234.x

The endocannabinoid anandamide (N‐arachidonoylethanolamine) and other bioactive long‐chain N‐acylethanolamines are thought to be formed from their corresponding N‐acylphosphatidylethanolamines by a specific phospholipase D (NAPE‐PLD) in the brain as well as other tissues. However, regional distribution of NAPE‐PLD in the brain has not been examined. In the present study, we investigated the expression levels of NAPE‐PLD in nine different regions of rat brain by enzyme assay, western blotting and real‐time PCR. The NAPE‐PLD activity was detected in all the tested brain regions with the highest activity in thalamus. Similar distribution patterns of NAPE‐PLD were observed at protein and mRNA levels. We also found a remarkable increase in the expression levels of protein and mRNA of the brain NAPE‐PLD with development, which was in good agreement with the increase in the activity. The age‐dependent increase was also seen with several brain regions and other NAPE‐PLD‐enriched organs (heart and testis). p‐Chloromercuribenzoic acid and cetyltrimethylammonium chloride, which inhibited recombinant NAPE‐PLD dose‐dependently, strongly inhibited the enzyme of all the brain regions. These results demonstrated wide distribution of NAPE‐PLD in various brain regions and its age‐dependent expression, suggesting the central role of this enzyme in the formation of anandamide and other N‐acylethanolamines in the brain.