Three to fifteen percent of peripheral T cells in adults express the recently described gamma delta T-cell antigen receptor (TcR) heterodimer. A small subpopulation of gamma delta cells express the CD8 accessory molecule. In this study, we analyzed the potential of highly purified CD4-/CD8-, double negative (DN) rat precursor thymocytes to give rise to gamma delta cells. We observed that in the presence of interleukin 2 (IL-2) and concanavalin A (ConA), both DN and CD8 cells expressing the gamma delta TcR were generated in vitro. We then examined the rat thymus for these cells and confirmed the presence of a previously undescribed CD8 TcR-alpha beta- subset in the rat thymus, expressing high levels of TcR-gamma and delta messages with no detectable TcR-alpha transcripts, similar to the cells generated in vitro in the presence of IL-2 and ConA.