Regional distribution and age-dependent expression of N-acylphosphatidylethanolamine-hydrolyzing phospholipase D in rat brain.
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.
Department of Biochemistry, Kagawa University School of Medicine, Kagawa, Japan.
Jun
J
Morishita
Yasuo
Y
Okamoto
Kazuhito
K
Tsuboi
Masaki
M
Ueno
Haruhiko
H
Sakamoto
Nobuhiro
N
Maekawa
Natsuo
N
Ueda
D000367:Age Factors; D000375:Aging / Q000502:physiology*; D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000033:anatomy & histology / Q000201:enzymology*; D001923:Brain Chemistry / Q000502:physiology; D019556:COS Cells; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000378:metabolism; D002522:Chlorocebus aethiops; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D008297:Male; D010714:Phosphatidylethanolamines / Q000235:genetics / Q000378:metabolism*; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013347:Subcellular Fractions / Q000201:enzymology; D020245:p-Chloromercuribenzoic Acid / Q000378:metabolism
D002593:Cetrimonium Compounds; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; D020245:p-Chloromercuribenzoic Acid; D010739:Phospholipase D; D000077286:Cetrimonium