The Leukocyte-specific adapter molecule SLP-76 (Src homology 2 domain-containing Leukocyte protein of 76 kilodaltons) is rapidly phosphorylated on tyrosine residues after receptor Ligation in several hematopoietically derived cell types. Mice made deficient for SLP-76 expression contained no peripheral T cells as a result of an early block in thymopoiesis. Macrophage and natural killer cell compartments were intact in SLP-76-deficient mice, despite SLP-76 expression in these lineages in wild-type mice. Thus, the SLP-76 adapter protein is required for normal-thymocyte development and plays a crucial role in translating signals mediated by pre-T cell receptors into distal biochemical events.