Secretin is a well-conserved member of the growth hormone-releasing hormone (GHRH) family of peptides expressed in brain, gut and gonads. To determine whether secretin may also play a physiological role in testis, we examined the level and cellular distribution of secretin and secretin receptor gene expression in rat testis. RNAs from total testis, Sertoli, germ and Leydig cells were amplified by comparative reverse transcription-polymerase chain reaction (RT-PCR). Southern blot analysis of the PCR products indicated secretin and secretin receptor mRNA expression primarily in germ cells. Sequence comparisons of cloned secretin and secretin receptor PCR products showed 100% identity with the previously reported sequences. To localize secretin and secretin receptor mRNAs at the cellular level within testis, in situ hybridization was performed. Specific hybridization to secretin mRNA was observed in low abundance in many germ cell types, but was heaviest over step 19 spermatids in stages VII and VIII tubules. Secretin receptor mRNA was detected in approximately the same cell types as was secretin mRNA, except that labeling was greatest in round spermatids (steps 6-8). Since the patterns of gene expression of secretin and its receptor overlap, these data suggest that there may be an intrinsic secretin system in testis.