Ryukyu spiny rats (genus Tokudaia), which are endemic to the central part of the Nansei Shoto archipelago in Japan, have unique karyotypes with odd numbers of chromosomes and no cytologically recognizable Y chromosome. The chromosome numbers of Tokudaia osimensis from Amamioshima and of Tokudaia sp. from Tokunoshima are 2n = 25 and 2n = 45, respectively, with a putative single X chromosome. The mouse X probe hybridized to the impaired X chromosome, except for the distal part of the short arm in a female specimen of T osimensis and in one male and one female of Tokudaia sp. Fluorescence in situ hybridization with the Tspy (testis-specific protein, Y-encoded) gene from both male and female cells of Tokudaia sp. by PCR localized Tspy to the distal part of the long arm of the X chromosome. Another Y-related gene, Zfy, from Tokudaia sp. was also localized to the same region in both species. Although the Sry gene is absent in this species, the present results suggest that the Y-chromosome segment carrying functional Y-linked genes, such as Tspy and Zfy, is translocated onto the distal part of the long arm of the X chromosome. Copyright (C) 2002 S. Karger AG, Basel.