Characterization and expression of a stage specific antigen by monoclonal antibody TRA 54 in testicular germ cells.

To study the mechanism of spermatogenesis, we have isolated many monoclonal antibodies (mAb) which recognize specific steps of mouse germ cell differentiation and then have evaluated the specific expression and characterization of antigenic molecules using immunohistochemistry and Western blotting. Monoclonal antibody TRA 54 recognized specific organelles in germ cell cytoplasm from spermatocytes to spermatids; that is, a large granule was stained in mid-late pachytene, diplotene and secondary spermatocytes and in round spermatids at stage I while the acrosome of spermatids at steps 2-3 to step 12 were also positive. Thereafter, the antigens disappeared from spermatids at more advanced stages of differentiation. Western blots using TRA 54 revealed broad bands with approximate molecular weights of >200, 190 and 85 kDa in the testis. The expression of these antigens during testicular germ cell development should be of interest in relation to the biogenesis of organelles such as the chromatoid body and acrosome and will be a useful stage-specific molecular marker for the study of spermatogenesis.