We found that immunoreactive endothelin (ET) is present in seminal fluid in very large amounts (500-5000 ng/L; quantification based on ET-1(1-21) standard). This immunoreactive ET was detected by use of a radioimmunoassay system in which the N-terminal portion of ET-1 and ET-2 (and big ET-1 and big ET-2) are recognized. Thus, the immunoreactive ET in seminal fluid may include the precursors of ET-1 or ET-2 (i.e., big ET) as well as metabolites of ET-1 or ET-2 in which the N-terminal region is intact. The levels of immunoreactive ET in seminal fluid from men with normal semen analyses and that in seminal fluid of vasectomized men were within the same range. Using a different radioimmunoassay system in which the C-terminal portion of ET-1, ET-2, and ET-3 is recognized, we found that the levels of immunoreactive ET were much lower (or undetectable). We speculate that bioactive ET may be produced and act to promote sperm transport in the male reproductive tract; thereafter, bioactive ET may be metabolized by membrane metalloendopeptidase (which is present in male reproductive tissues and semen) to immunoreactive, inactive products. Alternatively, big ET in seminal fluid may be processed in tissues of the female internal genitalia to bioactive ET, which could act to promote sperm transport through the uterine cavity by stimulating myometrial contractions.