Testatin is identified as a member of the Cystatin family and expressed in germ cells and somatic cells in reproductive tissues. Testatin transcription detectable in males and females at 9.5 clays Post coitum, before sex-differentiation, is up-regulated just after the onset of sry expression in the male gonads, while is clown-regulated to Undetectable levels in the female gonads. These expression patterns suggest that Testatin might have some roles in sex-differentiation. To address Testatin function in sex-differentiation, we analyzed the effects of ectopic-expression ill females and null-expression in males with testatin transgenic and knockout mice. In the transgenic females, testatin expression was constitutively elevated from embryonic gonad to adult ovary, and its expression was as high as the wild-type male gonads. However, both types of mice were fertile and did not exhibit detectable abnormalities. This Suggests that the decrease of endogenous testatin in female gonad is not critical, and the increase of testatin in male gonad is dispensable for sex-differentiation. (c) 2006 Elsevier Inc. All rights reserved.