Figure 3.
Long-chain ascarosides identified in the daf-22-mutant metabolome and interaction with NEA biosynthesis. (a) HPLC-MS analysis confirms long-chain ascarosides and ethanolamides in the daf-22 metabolome that are absent in wild-type. (b) daf-22 mutant-specific VLCAs identified via mvaDANS and subsequent HPLC-MS analysis. (c) Quantitative distribution of daf-22-upregulated ascarosyl methylketones. (d) Mutation of maoc-1, dhs-28, and daf-22 greatly reduces EPEA production and addition of ascarosides (2.5 μM ascr#3 and ascr#9, see reference (4) for structures) to daf-22(m130) cultures does not rescue EPEA production.* P<0.05, ** P<0.01, *** P<0.001. (e) Interactions of peroxisomal β-oxidation and endocannabinoid biosynthesis. Mutation of daf-22 abolishes processing of long-chain ascaroside CoA esters, whose conversion into ethanolamides is associated with reduced endocannabinoid production (13), likely due to depletion of phosphatidylethanolamine pools.