# Differential control of 2-AG’s activity at CB1R by ABHD6 and MAGL

> **NIH NIH R21** · UNIVERSITY OF WASHINGTON · 2024 · $233,250

## Abstract

Summary
The most abundant endocannabinoid (eCB) in brain, 2-arachidonoyl glycerol (2-AG), is inactivated by two
enzymes: monoacylglycerol lipase (MAGL) and α/β hydrolase domain-contain 6 (ABHD6) that differ in their
hydrolyzing activities and subcellular localization (presynaptic and postsynaptic, respectively). Accordingly,
selective inhibition of each enzyme results in different spatiotemporal enhancement of 2-AG-CB1R signaling in
the brain, and potentially synergistic therapeutic benefits.
 We recently gathered results showing that stimulated increase in 2-AG production is reliably measured using
GRABeCB2.0, a recently developed 2-AG sensor. Remarkably, metabotropic receptor mediated increase in 2-AG
is controlled by ABHD6, whereas ionotropic receptor-dependent increase in 2-AG is not controlled by ABHD6.
These results raise the question of how MAGL controls stimulation-dependent 2-AG production?
 Demonstrating that receptor-dependent increases in 2-AG production and activity at CB1R signaling are
differentially controlled by ABHD6 and MAGL would provide an additional level of mechanistic distinction
between these eCB-hydrolyzing enzymes. To increase our understanding of the respective roles of ABHD6 and
MAGL in controlling 2-AG-CB1R signaling in brain, we initiated an effort and have now successfully validated the
GRABeCB2.0 pharmacological profile in neural cells in culture and identified several stimuli that increase 2-AG
production. Based on this premise, we propose to address the following two questions in mouse neurons in
primary culture and striatal slices using live-cell fluorescence microscopy, two-photon microscopy and
electrophysiology:
Aim 1: Which stimuli increase 2-AG production and GRABeCB2.0 signal in neurons?
Aim 2: How do ABHD6 and MAGL differentially control stimuli-dependent increases in 2-AG-CB1R
signaling in neurons?
Completion of the work outlined in this new R21 grant proposal will provide a comprehensive understanding of
the respective role of ABHD6 and MAGL in controlling 2-AG-CB1R signaling in the brain. Our long-term goal is
to increase our understanding of the molecular, cellular, and system’s level differences by which ABHD6 and
MAGL control eCB signaling in brain, a body of work that will help develop novel therapeutics with reduced
potential for abuse and adverse effects produced by classic cannabinoid agonists.

## Key facts

- **NIH application ID:** 10814943
- **Project number:** 5R21DA056816-02
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Nephi Stella
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $233,250
- **Award type:** 5
- **Project period:** 2023-04-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10814943

## Citation

> US National Institutes of Health, RePORTER application 10814943, Differential control of 2-AG’s activity at CB1R by ABHD6 and MAGL (5R21DA056816-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10814943. Licensed CC0.

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