The PDE1A gene encodes a Ca2+/calmodulin-stimulated 3',5'-cyclic nucleotide phosphodiesterase (PDE). We have performed 5' and 3' RACE and identified two additional 5'-splice variants and one additional 3'-splice variant of the human PDE1A gene. The three known 5'-splice variants and the two known 3'-splice variants combine to generate six different PDE1A mRNAs. However, one of the 5'-splice variants exhibits alternate splicing in the 5' untranslated region. Thus the six mRNAs encode four different PDE1A proteins. Recombinant forms of the different human PDE1A isoforms were expressed in Sf9 cells. The kinetic properties and inhibitor sensitivities of the four PDE1A isoforms are very similar to one another.