Dietary manipulation produces marked alterations in desaturase activities of rat liver microsomes with no concomitant changes in acyltransferase activities. Desaturation of stearoyl-CoA (delta 9-desaturase), linoleoyl-CoA (delta 6-desaturase), eicosatrienoyl-CoA (delta 5-desaturase) and eicosatrienoyl-phosphatidylcholine (delta 5-desaturase) was elevated in animals fed a corn oil diet and lowered in those fed a coconut oil diet compared to control animals. The delta 5-desaturase activities were also lowered in starved animals and elevated in starved animals refed a fat-free diet. However, no changes in acyl-CoA:1-acyl-sn-glycero-3-phosphocholine acyltransferase activity were observed in the membranes of animals maintained on any of the dietary regimens studied. These observations suggest that the desaturases of rat liver microsomes are regulated independently of the acyltransferases and that desaturation of eicosatrienoyl-phosphatidylcholine is regulated at the level of the desaturase itself and not by availability of the phospholipid substrate.