The existence of a third non-coding exon in the human neprilysin-encoding gene (h-NEP), positionally located as exon 3, has been demonstrated by reverse transcription of RNA from human kidney and lung, coupled with the polymerase chain reaction. Comparison of nucleotide sequences between h-NEP and the rat NEP (r-NEP) genes shows a high degree of sequence conservation within noncoding exons 1 and 2. In contrast, the region of the gene containing exon 3 is highly divergent. Two transcripts derived from exon 2 by alternative splicing, type-2a and type-2b, were demonstrated in human kidney and lung. In contrast, only the type-2b transcript was present in these same tissues in the rat. The type-1 transcript was detected in human kidney, lung and brain, this transcript appearing to be the major species in brain.