PC4 is a serine protease primarily expressed in spermatids. We have produced PC4-deficient mice carrying an insertion of the bacterial gene for beta galactosidase under the PC4 gene promoter. Male mice lacking PC4 (-/-) exhibit severely reduced fertility. Surprisingly, the fertility of female mice is also significantly diminished in these mutants (Mbikay et al., 1997. Proc. Natl. Acad. Sci. USA 94, 6842-6846). The aim of this study was to determine the site of PC4 expression in mouse ovaries. Using a histoenzymatic assay for beta-galactosidase, we show that the PC4 promoter can drive strong expression of this enzyme in the theca-interstitium and in degenerating corpora lutea of +/- ovaries. We also demonstrate that PC4 transcripts can be detected by RT-PCR in the ovaries of +/- and +/+ mice, but not in those of -/- mice. The cells expressing PC4 were macrophage-like, since they expressed the macrophage markers CD11b and F4/80, as well as interleukin 1beta and tumor necrosis factor alpha (TNFalpha). Expression of PC4 was also detected in the mouse macrophage RAW264.7 cell line. Interestingly, TNFalpha transcripts were 3-fold more abundant in ovarian macrophage-like cells from -/- mice than in those from +/+ mice, suggesting a constitutive state of activation of the mutant cells. An inverse relationship between PC4 expression and macrophage activation was also observed in RAW264.7 cells. When these cells were activated using bacterial lipopolysaccharide, the level of PC4 transcripts decreased, while that of TNFalpha increased. These observations identify PC4 as an enzyme that could influence ovarian physiology by affecting macrophage functions.