The Prnp (prion protein) gene, which encodes a soluble protein anchored to the cell surface by glycosylphosphatidylinositol (GPI), might be involved in cell-to-cell interaction. The expression of Prnp is strongly observed not only in the brain, but also in non-neuronal tissues. In order to examine the Prnp expression sites in mouse testes, we carried out Northern blot and in situ hybridization analyses. By Northern blot analysis, two kinds of Prnp transcripts (major band of 2.2 kb, and minor band of 1.1 kb) were detected in testes. The 2.2-kb transcript was observed in testes throughout the postnatal development, whereas the 1.1-kb transcript was observed in testes from 2 to 70 weeks old. In situ hybridization analysis showed that the positive signals for Prnp mRNAs were predominantly observed in spermatogenic cells, but not in somatic cells such as Sertoli cells, Leydig cells and peritubular myoid cells. The signals were observed moderately in spermatogonia, and strongly in spermatocytes and round spermatids, but not in elongate spermatids and spermatozoa. These results suggest that Prnp may be involved in germ cell differentiation during mammalian spermatogenesis.