Measurement of Cellular Diacylglycerol Content

With the discovery of the role of polyphosphoinositide hydrolysis in mediating the action of calcium-mobilizing hormones, growth factors, and neurotransmitters (reviewed in ref. 1 ), the development of methods for measuring the formation of the resultant second messengers, inositol 1,4,5-trisphosphate and diacylglycerol (DAG), became essential. The additional discovery of an effector enzyme, protein kinase C, for transducing the changes in DAG levels (reviewed in ref. 2 ) underlined the importance of this second messenger. A sensitive and reliable protocol currently in use in many laboratories is described below for the measurement of DAG levels in stimulated cells.