Two mammalian isoforms of phospholipase D, PLD1 and PLD2, have recently been characterized at the molecular level. Effects of physiologic agonists on PLD activity in intact cells, as characterized in earlier studies, have generally not been attributed to specific PLD isoforms. Recent work has established that expression of PLD1 and PLD2 varies within tissues and between cell lines. A single cell type can express one, both, or neither isoform, although most cells co-express PLD1 and PLD2. Lymphocytes often lack expression of one or both isoforms of PLD. Relative levels of PLD mRNA expression vary considerably between established cell lines. Expression of transcripts for both PLD1 and PLD2 can be regulated at the transcriptional level by growth and differentiation factors in cultured cells. Thus, it is apparent that the known mammalian PLD isoforms are subject to regulation at the transcriptional level. The available data do not conclusively establish whether PLD1 and PLD2 are the only isoforms responsible for agonist-mediated PLD activation. Further studies of the regulation of expression of PLD isoforms should provide insight into the roles of PLD1 and PLD2 in physiologic responses, and may suggest whether additional forms of PLD remain to be characterized.