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In clinically healthy cows, lipolysis intensity is reduced throughout lactation; however, if FFA release exceeds tissue demands or the liver\u2019s metabolic capacity, lipid byproducts accumulate, increasing cows\u2019 risk of metabolic and infectious disease. Endocannabinoids (eCBs) and their congeners,\n                      <jats:italic>N-<\/jats:italic>\n                      acylethanolamines (NAEs), are lipid-based compounds that modulate metabolism and inflammation. Their synthesis and release depend upon the availability of FFA precursors and the abundance of synthesizing and degrading enzymes and transporters. Therefore, we hypothesized that eCB production and transcription of endocannabinoid system components are modulated by lipolysis pathways in adipocytes. To test this hypothesis, we stimulated canonical (isoproterenol, 1\u00a0\u00b5mol\/L; ISO) and inflammatory (lipopolysaccharide, 1\u00a0\u00b5g\/mL; LPS) lipolysis pathways in adipocytes isolated from the AT of 5 Holstein dairy cows. Following, we assessed lipolysis intensity, adipocytes\u2019 release of eCBs, and transcription of endocannabinoid system components.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      We found that ISO and LPS stimulated lipolysis at comparable intensities. Exposure to either treatment tended to elevate the release of eCBs and NAEs by cultured adipocytes; however, specific eCBs and NAEs and the transcriptional profiles differed by treatment. On one hand, ISO enhanced adipocytes\u2019 release of 2-arachidonoylglycerol (2-AG) but reduced NAE production. Notably, ISO enhanced the cells\u2019 expression of enzymes associated with 2-AG biosynthesis (\n                      <jats:italic>INPP5F<\/jats:italic>\n                      ,\n                      <jats:italic>GDPD5<\/jats:italic>\n                      ,\n                      <jats:italic>GPAT4<\/jats:italic>\n                      ), transport (\n                      <jats:italic>CD36<\/jats:italic>\n                      ), and adipogenesis (\n                      <jats:italic>PPARG<\/jats:italic>\n                      ). Conversely, LPS enhanced adipocytes\u2019 synthesis and release of\n                      <jats:italic>N-<\/jats:italic>\n                      arachidonoylethanolamide (AEA). This change coincided with enhanced transcription of the NAE-biosynthesizing enzyme,\n                      <jats:italic>PTPN22<\/jats:italic>\n                      , and adipocytes\u2019 transcription of genes related to eCB degradation (\n                      <jats:italic>PTGS2<\/jats:italic>\n                      ,\n                      <jats:italic>MGLL<\/jats:italic>\n                      ,\n                      <jats:italic>CYP27B1<\/jats:italic>\n                      ). Furthermore, LPS enhanced adipocytes\u2019 transcription of eCB and NAE transporters (\n                      <jats:italic>HSPA1A<\/jats:italic>\n                      ,\n                      <jats:italic>SCP2<\/jats:italic>\n                      ) and the expression of the anti-adipogenic ion channel,\n                      <jats:italic>TRPV3<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our data provide evidence for distinct modulatory roles of canonical and inflammatory lipolysis pathways over eCB release and transcriptional regulation of biosynthesis, degradation, transport, and ECS signaling in cows\u2019 adipocytes. Based on our findings, we conclude that, within adipocytes, eCB production and ECS component expression are, at least in part, mediated by lipolysis in a pathway-dependent manner. These findings contribute to a deeper understanding of the molecular mechanisms underlying metabolic regulation in dairy cows\u2019 AT, with potential implications for prevention and treatment of inflammatory and metabolic disorders.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s40104-024-01062-z","type":"journal-article","created":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T20:02:04Z","timestamp":1722628924000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Lipolysis pathways modulate lipid mediator release and endocannabinoid system signaling in dairy cows\u2019 adipocytes"],"prefix":"10.1186","volume":"15","author":[{"given":"Madison N.","family":"Myers","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel","family":"Chirivi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeff C.","family":"Gandy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Tam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maya","family":"Zachut","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4969-2178","authenticated-orcid":false,"given":"G. 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