Identification of gene targets of the transcription factor SALL4 in undifferentiated spermatogonia

Sustained spermatogenesis in adult males and recovery of fertility following germ cell depletion are dependent on undifferentiated spermatogonia with self-renewal potential. We have previously demonstrated a critical role for the transcription factor Spalt-like 4 (SALL4) in spermatogonial differentiation. However, it remains unclear whether SALL4 has broader roles within the spermatogonial pool despite its ability to co-regulate genes with PLZF, a transcription factor required for undifferentiated cell maintenance. To identify genes regulated by SALL4 in the male germline, we established cultures of undifferentiated spermatogonia from a Sall4 inducible knockout mouse model. Cells were treated with vehicle (as control) or tamoxifen to induce gene deletion, then cells harvested and analysed by microarray to identify genes mis-expressed upon loss of SALL4. Three independently derived Sall4flox/flox Ubc-CreER (Sall4TAM-KO) cultured undifferentiated spermatogonia were treated with vehicle (CTRL) or 4-hydroxytamoxifen (TAM) and harvested 4 days later.