Inactivation of one of the 2 X chromosomes in the somatic cells of female mammals is the process by which their X-linked gene products are equalized to those of their male counterparts. In male mammals, however, a sex vesicle representing the condensed and transcriptionally silenced sex chromosomes is detected during early meiotic prophase. Since the exact stage of development at which X inactivation is initiated in the bovine testis is not established as yet, we undertook to study fetuses ranging in age from 30 to 180 days of gestation, to determine the transcriptional status of the Xist gene currently thought to be the prerequisite component of X inactivation. Our studies using reverse transcription polymerase chain reaction (RT-PCR) approach with primers designed to amplify a 463 bp product from a conserved region of the first exon of bovine Xist gene, proved that Xist expression is evident in bovine fetal testes as early as 50 days of gestation and that it continues at least to the end of the second trimester (180 days) of gestation. Morphological studies on fetal testes during gestational stage spanning the period of Xist expression revealed the presence of large intra-tubular cells overtly resembling the prespermatogonia of postnatal bovine testes, at 50 days and preleptotene like cells as early as 90 days of gestation. We hypothesize that the expression of the Xist gene, or the recently discovered Tsix gene antisense to Xist in orientation, may be related to the presence of these cells which participate in the morphogenesis of the fetal bovine testis.