{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T21:03:34Z","timestamp":1789506214198,"version":"build-2803163510"},"reference-count":56,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["AT011162"],"award-info":[{"award-number":["AT011162"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2026,9]]},"abstract":"\n Abstract<\/jats:title>\n \n The endogenous cannabinoid signalling system consists of G protein\u2010coupled receptors, messengers \u2013 2\u2010arachidonoylglycerol (2\u2010AG) and anandamide \u2013 and enzymatic machinery to synthesize and metabolize these messengers. Anandamide is physiologically important, but its synthesis is incompletely understood.\n N<\/jats:italic>\n \u2010Arachidonoyl\u2010phosphatidylethanolamide phospholipase D (NAPE\u2010PLD) synthesizes acylethanolamines, including anandamide, but how is NAPE\u2010PLD activated? We used genetically encoded G protein\u2010coupled receptor based (GRAB) sensors for endocannabinoids (eCBs) and immunohistochemistry to investigate. Human embryonic kidney 293 (HEK293) cells natively express G\n q<\/jats:sub>\n \u2010coupled muscarinic M\n 3<\/jats:sub>\n receptors. The muscarinic agonist oxotremorine\u2010M stimulated the GRAB\n eCB<\/jats:sub>\n sensor only when HEK293 cells were cotransfected with NAPE\u2010PLD. This signal was reduced by the NAPE\u2010PLD inhibitor LEI401 and required both phospholipase C and internal calcium stores. G\n q<\/jats:sub>\n \u2010coupled mGluR5 glutamate receptors effectively substitute for M\n 3<\/jats:sub>\n receptors. Thus M\n 3<\/jats:sub>\n receptors stimulate NAPE\u2010PLD synthesis of acylethanolamines such as anandamide via G\n q<\/jats:sub>\n signalling pathways resembling those for 2\u2010arachidonoylglycerol. Parasympathetic activation stimulates tearing and salivation via M\n 3<\/jats:sub>\n receptors on myoepithelial cells. The cannabinoid signalling system may act as a feedback inhibitor to inhibit acetylcholine release, but the identity and source of the endogenous cannabinoid messenger are uncertain. NAPE\u2010PLD and M\n 3<\/jats:sub>\n proteins colocalize in myoepithelial cells; fatty acid amide hydrolase (FAAH) resides in glandular acinar cells. Oxotremorine\u2010M stimulation of lacrimal or submandibular salivary glands co\u2010cultured with HEK293\u2010GRAB\n eCB<\/jats:sub>\n cells stimulated GRAB\n eCB<\/jats:sub>\n responses. This response was diminished by LEI401 and was also present in cells expressing GRAB\n AEA<\/jats:sub>\n , an anandamide\u2010specific sensor, but not those expressing GRAB\n 2AG<\/jats:sub>\n . We conclude that muscarinic M\n 3<\/jats:sub>\n receptors stimulate NAPE\u2010PLD to produce anandamide in cell lines and exocrine glands. This means of stimulating anandamide synthesis may play many roles in the body.\n \n \n image<\/jats:alt-text>\n <\/jats:graphic>\n <\/jats:boxed-text>\n <\/jats:p>\n <\/jats:sec>\n \n Key points<\/jats:title>\n \n \n \n The cannabinoid signalling system plays important roles in the body and in doing so makes use of two endogenous messengers, 2\u2010arachidonoylglycerol (2\u2010AG) and anandamide. The regulation of anandamide synthesis is still poorly understood.<\/jats:p>\n <\/jats:list-item>\n \n \n This study demonstrates that muscarinic G\n q<\/jats:sub>\n \u2010coupled GPCRs can stimulate anandamide production by activating the enzyme\n N<\/jats:italic>\n \u2010arachidonoyl\u2010phosphatidylethanolamide phospholipase D (NAPE\u2010PLD) and further shows that this occurs natively in salivary and lacrimal glands.\n <\/jats:p>\n <\/jats:list-item>\n \n \n Muscarinic G\n q<\/jats:sub>\n \u2010coupled G protein\u2010coupled receptor (GPCR) activation of NAPE\u2010PLD to induce anandamide synthesis may play many roles in the body.\n <\/jats:p>\n <\/jats:list-item>\n <\/jats:list>\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1113\/jp291508","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T04:20:25Z","timestamp":1788409225000},"page":"7721-7738","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["N<\/i>\n \u2010Arachidonoyl\u2010phosphatidylethanolamide phospholipase D activation by muscarinic receptors"],"prefix":"10.1113","volume":"604","author":[{"given":"Natalia","family":"Murataeva","sequence":"first","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Connor","family":"Schmitt","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elyssa","family":"Hyman","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jim","family":"Wager\u2010Miller","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Matthews","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruyi","family":"Cai","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Membrane Biology Peking University School of Life Sciences Beijing China"},{"name":"PKU\u2010IDG\/McGovern Institute for Brain Research Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shangxuan","family":"Cai","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Membrane Biology Peking University School of Life Sciences Beijing China"},{"name":"PKU\u2010IDG\/McGovern Institute for Brain Research Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulong","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Membrane Biology Peking University School of Life Sciences Beijing China"},{"name":"PKU\u2010IDG\/McGovern Institute for Brain Research Beijing China"},{"name":"Peking\u2010Tsinghua Center for Life Sciences, New Cornerstone Science Laboratory Academy for Advanced Interdisciplinary Studies Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ken","family":"Mackie","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0567-5317","authenticated-orcid":false,"given":"Alex","family":"Straiker","sequence":"additional","affiliation":[{"name":"Department of Psychological and Brain Sciences Indiana University Bloomington Indiana USA"},{"name":"Gill Institute for Neuroscience Indiana University Bloomington Indiana USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-17987-2"},{"issue":"1","key":"e_1_2_5_3_1","first-page":"14","article-title":"Expression of G protein\u2010coupled receptors and related proteins in HEK293, AtT20, BV2, and N18 cell lines as revealed by microarray analysis","volume":"12","author":"Atwood B. 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