Current evidence suggests the presence of transcripts for several secretory phospholipase A(2) (sPLA(2)) enzymes in male genital organs. In this study, we examined by immunohistochemistry the localization of group IIA, IIC, IID, IIE, IIF, V and X sPLA(2)s in male genital organs. In sPLA(2)-IIA-deficient C57BL/6 mouse testis, sPLA(2)-IIC, -IID, -IIE, -IIF, -V and -X were diversely expressed in spermatogenic cells within the seminiferous tubules. Immunoblotting revealed the presence of these sPLA(2)s in mouse spermatozoa. In addition, sPLA(2)-IIF, -V and -X were localized in the interstitial Leydig cells. The same set of sPLA(2)s was detected in a mouse cultured Leydig cell line, and adenovirus-mediated transfer of these sPLA(2)s into Leydig cells resulted in increased prostaglandin production. sPLA(2)-IIC, -IID, -IIE, -IIF, -V and -X were also detected diversely in the epithelium of the epididymis, vas deferens, seminal vesicles, and prostate. In a sPLA(2)-IIA-positive FVB strain, weak expression of sPLA(2)-IIA was detected in Leydig cells. Notable differences in the sPLA(2) expression profiles were found in the seminal vesicles and prostate between mice and humans. Taken together, individual sPLA(2)s exhibit distinct or partially overlapping localizations in male reproductive organs, suggesting both specific and redundant functions.