AbstractOne of the principal monoacylglycerol (MAG) species in animal tissues is 2‐arachidonoylglycerol (2‐AG), and the diacylglycerol lipase (DAGL) pathway is the most important 2‐AG biosynthetic pathway proposed to date. Lysophosphatidate phosphatase (LPAase) activity is part of another 2‐AG‐forming pathway in which monoacylglycerol lipase (MAGL) is the major degrading enzyme. The purpose of this study was to analyze the manner in which DAGL, LPAase, and MAGL enzymes are modified in the central nervous system (CNS) during aging. To this end, diacylglycerols (DAGs) and MAGs of different composition were used as substrates of DAGL and MAGL, respectively. All enzymatic activities were evaluated in membrane and soluble fractions as well as in synaptic terminals from the cerebral cortex (CC) of adult and aged rats. Results related to 2‐AG metabolism show that aging: (a) decreases DAGL‐α expression in the membrane fraction whereas in synaptosomes it increases DAGL‐β and decreases MAGL expression; (b) decreases LPAase activity in both membrane and soluble fractions; (c) decreases DAGL and stimulates LPAase activities in CC synaptic terminals; (d) stimulates membrane‐associated MAGL‐coupled DAGL activity; and (e) stimulates MAGL activity in CC synaptosomes. Our results also reveal that during aging the net balance between the enzymatic activities involved in 2‐AG synthesis and breakdown is low availability of 2‐AG in CC membrane fractions and synaptic terminals. Taken together, our results lead us to conclude that these enzymes play crucial roles in the regulation of 2‐AG tissue levels during aging. © 2012 BioFactors, 2013