Gene expression analysis of mouse testis after treatment with histone deacetylase inhibitor TSA

Trichostatin A (TSA) is one of the most potent reversible histone deacetylase (HDAC) inhibitors. In male mice, subcutaneous application of TSA is followed by infertility due to apoptosis of pachytene spermatocytes. To get more insight into the mechanisms underlying this phenomenon, we performed a genome-wide expression analysis of murine testes after TSA treatment. The whole genome transcriptome analysis revealed 507 significantly regulated genes. Validation by real-time quantitative PCR confirmed the expression of 7 from 9 genes tested (78%). Keywords: time course TSA was resolved in dimethylsulfoxide and PBS. 15 animals were divided into 5 groups each containing 3 mice. Group 1 received only the carrier and were sacrificed after 10h (control). Groups 2 - 5 received 60ug/day/mouse TSA applied by one subcutaneous injection and were sacrificed after 2.5h (group 2), 5h (group 3), 7.5h (group 4) and 10h (group 5).Total RNA was extracted from frozen tissue of 3 animals per group. The variability between animals in each group was minor, as demonstrated by the histological and immunohistochemical stainings. Therefore, the material from each group was pooled. cRNA of each group was hybridized in duplicate (technical replicates). In total, 10 bioarrays (2 arrays per group) were processed in parallel.