Spatially Resolved Mapping of Monoacylglycerol Lipase Activity in the Brain

van der Vliet, Daan and Klinkenberg, Alex X.Y. and Platte, Rik and Higgins, Kieran and Prokop, Susanne Clara and Huizenga, Mirjam C.W. and Kraaijevanger, Lars and van Egmond, Noëlle and Straub, Verena M. and Kole, Maarten H.P. and Pacher, Pál and Katona, István and Huitinga, Inge and van der Stelt, Mario (2025) Spatially Resolved Mapping of Monoacylglycerol Lipase Activity in the Brain. ACS CHEMICAL NEUROSCIENCE, 16 (24). pp. 4622-4635. ISSN 1948-7193

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Abstract

Visualizing signaling systems in the brain with high spatial resolution is critical to understanding brain function and to develop therapeutics. Especially, enzymes are often regulated on the post-translational level, resulting in a disconnect between protein levels and activity. Conventional antibody-based methods have limitations, including potential cross-reactivity and the inability of antibodies to discriminate between active and inactive enzyme states. Monoacylglycerol lipase (MAGL), an enzyme degrading the neuroprotective endocannabinoid 2-arachidonoylglycerol, is the target of inhibitors currently in clinical trials for the treatment of several neurological disorders. To support translational and (pre)clinical studies and fully realize the therapeutic opportunities of MAGL inhibitors, it is essential to map the spatial distribution of MAGL activity throughout the brain in both health and disease. Here, we introduce selective fluorescent activitybased probes for MAGL enabling direct visualization of its enzymatic activity in lysates, cultured cells, and tissue sections. We show that oxidative stress, which inactivates MAGL through the oxidation of regulatory cysteines, reduces probe labeling, thereby validating the probes activity-dependence. Extending this approach, we developed an activity-based histology protocol to visualize MAGL activity in fresh-frozen mouse and human brain tissues. This approach revealed robust MAGL activity in astrocytes and presynaptic terminals within the mouse hippocampus and further allows detection of MAGL activity in the human cerebral cortex. Collectively, these findings establish selective activity-based probes as powerful tools mapping MAGL activity with high spatial resolution across mammalian brain tissue.

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