Inhalation of diesel exhaust particles (DEP) has been found to reduce sperm production. There is a possibility that DEP exposure elicits changes in gene expression in testis. To identify the alteration of gene expression resulting from DEP exposure, we constructed subtracted cDNA libraries from mouse testis using the suppression subtractive hybridization method. We isolated 16 candidate clones whose expression levels changed after exposure. Some of these candidates were highly similar to known testis-specific genes. Some of the clones also seemed to correlate with spermatogenesis. Northern blot analysis revealed that DEP exposure changed the expression levels of several clones in a dose-dependent manner. For example, the expression of clone R8, which was very similar to human XRRA1, increased by 2.3-fold in testis after DEP exposure. On the other hand, the expressions of long-chain acyl-CoA synthetase 6 (Acsl6) and serine/threonine kinase 35 reduced by 0.3-fold. These results indicated that some constituents of DEP alter gene expression in the testis.