γH2AX marks the main events of the spermatogenic process

Abstract

Recent findings reveal that chromatin reorganization requires formation and repair of DNA double strand breaks (DSBs). Therefore, dynamic changes in chromatin structure taking place along the mitotic and the meiotic processes should be accompanied by histone H2AX phosphorylation and formation of gamma H2AX, a prominent event occurring at DSB sites. Accordingly, gamma H2AX has been detected at multiple developmental steps in adult germ cells. Nevertheless, no comprehensive study focusing on the main biological events occurring in the seminiferous epithelium has been carried out in mammals. Here I analyze H2AX phosphorylation in immunostained testis sections comparing PAS/cresyl violet counterstained, noncounterstained, and immuno-fluorescence preparations and show several waves of H2AX phosphorylation/dephosphorylation coupled to various developmental phases of spermatogonia and spermatocytes as well as to spermatid differentiation. The combined detection of DNA replication or free 3'-OH DNA ends highlight the possible significance of gamma H2AX at these locations.