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N -acylethanolamines are an important class of lipid signaling molecules found in many species, including the nematode  Caenorhabditis elegans  ( C. elegans ) where they are involved in development and adult lifespan. In mammals, the relative activity of the biosynthetic enzyme  N -acyl phosphatidylethanolamine-specific phospholipase-D and the hydrolytic enzyme fatty acid amide hydrolase determine  N -acylethanolamine levels.  C. elegans  has two  N -acyl phosphatidylethanolamine-specific phospholipase-D orthologs,  nape-1  and  nape-2 , that are likely to have arisen from a gene duplication event. Here, we find that recombinant  C. elegans  NAPE-1 and NAPE-2 are capable of generating  N -acylethanolamines  in vitro , confirming their functional conservation.  In vivo , they exhibit overlapping expression in the pharynx and the nervous system, but are also expressed discretely in these and other tissues, suggesting divergent roles. Indeed,  nape-1  over-expression results in delayed growth and shortened lifespan only at 25°C, while  nape-2  over-expression results in significant larval arrest and increased adult lifespan at 15°C. Interestingly, deletion of the  N -acylethanolamine degradation enzyme  faah-1  exacerbates  nape-1  over-expression phenotypes, but suppresses the larval arrest phenotype of  nape-2  over-expression, suggesting that  faah-1  is coupled to  nape-2 , but not  nape-1 , in a negative feedback loop. We also find that over-expression of either  nape-1  or  nape-2  significantly enhances recovery from the dauer larval stage in the insulin signaling mutant  daf-2(e1368) , but only  nape-1  over-expression reduces  daf-2  adult lifespan, consistent with increased levels of the  N -acylethanolamine eicosapentaenoyl ethanolamine. These results provide evidence that  N -acylethanolamine biosynthetic enzymes in  C. elegans  have conserved function and suggest a temperature-dependent, functional divergence between the two isoforms.

Characterization of N-Acyl Phosphatidylethanolamine-Specific Phospholipase-D Isoforms in the Nematode Caenorhabditis elegans