Most murine thymocytes and mature T cells originate from a numerically minor population of CD8-4- (double-negative, DN) thymocytes. In this study, we investigated the effects of rearranged T cell receptor (TcR) alpha and beta transgenes on early T cell development. We analyzed the precursor potential, the expression of CD25 and TcR at mRNA and/or protein level in DN thymocyte subsets in TcR transgenic (Tg) mice. We report the following observations: (i) despite a large overrepresentation of total DN cells in TcR Tg mice, precursor-containing CD25+ DN and CD8lo4lo thymocytes are reduced to a third of the nontransgenic control numbers; (ii) like in the normal mice, CD25+ DN and CD8lo4lo can, and TcR+ DN cells cannot generate other thymic subsets; (iii) TcR alpha mRNA and TcR alpha/beta protein levels are quantitatively increased, but their developmental expression is similar to that in normal mice; and (iv) surface TcR alpha beta expression becomes detectable as the thymocytes down-regulate CD25, paralleling the situation in normal mice. Our findings implicate stringent transcriptional control, rather than TcR gene rearrangement, as a decisive regulator of TcR alpha beta expression in early ontogeny.