The tree mouse Vandeleuria o. oleracea has an odd diploid chromosome complement (2n = 29 female/male) accompanied by a unique multiple sex-chromosome mechanism (X1X2Ymale/X1X1X2female). In the present paper the sex chromosomes have been identified unequivocally with the help of G-banding and DNA replication patterns which confirm our earlier suggestion that the X1 is smaller in size than the simple mammalian X, and also that the combined sizes of the X1 and X2 (3-8 and 1-7 per cent respectively) approximate to the size of the conservative mammalian X (5 per cent). It is proposed that the mammalian X has attained a minimum functional size so that further reduction is not tolerated.