Investigation of endocannabinoid 2-AG and its hydrolases for regulation on sperm motility and acrosome reaction in Kunming mice

CONTEXT: A key endocannabinoid, 2-arachidonoyl glycerol (2-AG), regulates sperm hyperactivation in humans via alpha/beta hydrolase domain-containing protein 2 (ABHD2)-mediated CatSper calcium channel activation. However, the role of ABHD2 in mice is unclear, whereas monoacylglycerol lipase (MAGL), ABHD-containing protein 6 (ABHD6), and ABHD-containing protein 12 (ABHD12) modulate 2-AG in the mouse central nervous system (CNS). AIMS: This study explored the impact of 2-AG on mouse sperm motility, its regulatory mechanisms, and the involvement of MAGL, ABHD6, and ABHD12. METHODS: 2-AG concentrations in caput and caudal epididymal sperm from Kunming mice were quantified. ABHD2/6/12 and MAGL were identified, localized, and assessed for roles in motility and calcium (Ca2+) influx. KEY RESULTS: Caput sperm exhibited higher 2-AG contents than did caudal sperm. Four hydrolases regulated motility via 2-AG hydrolysis to induce calcium influx. MAGL showed the strongest hydrolytic activity, ABHD2 the weakest. Subcellular localization showed MAGL in mid-piece, ABHD12 in mid-piece and principal piece, and ABHD2/ABHD6 in acrosomal region. CONCLUSIONS: 2-AG modulates mouse sperm motility via calcium signaling. MAGL, ABHD6, and ABHD12 drive 2-AG hydrolysis, while ABHD2 is linked to acrosome exocytosis, not 2-AG breakdown. IMPLICATIONS: Targeting MAGL/ABHD12 offers non-hormonal contraceptive strategies or assisted reproductive technology (ART) improvements. Monitoring 2-AG and hydrolase activity could aid in diagnosing male infertility.