Demonstration of terminal deoxynucleotidyl transferase in thymocytes by immunofluorescence

Abstract

The cellular and subcellular distribution of terminal deoxynucleotidyl transferase (DNA nucleotidylexotransferase; nucleosidetriphosphate:DNA deoxynucleotidylexotransferase, EC 2.7.7.31) in thymocytes and peripheral lymphocytes from rat, mouse, and calf was studied by immunofluorescence using rabbit antiserum to homogeneous transferase from calf. Terminal transferase was readily detected in approximately 75% of cortical thymocytes, but not in medullary thymocytes or lymph node lymphocytes. The enzyme appeared to be present predominantly in the cytoplasm of positive thymocytes in ethanol-fixed cell smears and frozen sections. The reactivity of anti-terminal-transferase for thymocytes could be neutralized with purified calf enzyme. Results of experiments in which thymocytes were separated on 7-step discontinuous Ficoll density gradients suggested that cortical thymocytes are heterogeneous with respect to terminal deoxynucleotidyl transferase content.