Although the athymic nude mouse is grossly deficient in peripheral T cells, the number of lymphocytes bearing T-cell markers (L3T4, LyT2) and the alpha beta or gamma delta T-cell receptor (Tcr) increases steadily with age. The anatomical site(s) where these cells arise are unknown. Splenocytes from 3-5-week-old C57BL/6 (nu/nu) mice contain 2%-5% Pro-T cell progenitors identified with the Joro 37-5 and Joro 75 antibodies, but not mature T cells. To study Tcr gene rearrangement outside the thymus, we fused splenocytes from 3-5-week-old C57BL/6 nude mice with the T-cell lymphoma BW 100.129. Of 22 hybrids that grew stably in culture, four had Tcrd-VD1-D2-J1, two had Tcrd-VD2-J1, and seven had Tcrd-D1-D2 types of rearrangement. Eight hybrids had rearranged the Tcrg-2 gene cluster, but none had rearranged Tcrg-1, -3, or -4. None of the hybrids had rearranged the Tcrb gene cluster and 13 contained DJ rearrangements at the Igh locus. We conclude that the spleen is one of the extrathymic sites where T-cell progenitors can rearranged Tcrd and Tcrg genes. However, there was no evidence for Tcrb gene rearrangements in this organ. Furthermore, the analysis of this limited number of hybrids suggests that extrathymic Tcr gene rearrangements seem to be distinct and much less diverse than those found in the developing thymocytes.