Transcriptome profiling of OCT4 positive spermatogonia in wild type and Akt1s1 null mutant mice.

In order to investigate the regulatory roles of AKT1S1 during mouse spermatogenesis, we analyzed gene expression changes that caused by the deletion of Akt1s1 gene in mouse male germ cells using RNA sequencing method. The transcriptome profiling analyses showed that OCT4+ spermatogonia from mice at different ages possess varied changes in gene expression due to the deletion of Akt1s1 gene, supporting the role of AKT1S1 plays in mediating AKT-mTORC1 signaling during mouse spermatogenesis. Overall design: Spermatogonia expressing OCT4 were isolated from mouse testes expressing green fluorescent protein (GFP) under the control of Pou5f1 (the gene encoding OCT4) promoter in either wild type or Akt1s1 null mutant background. OCT4+ cells were FACS isolated from enzyme-dispersed testicular cells using mice at three time points (3 days, 7 days and 2-3 months following birth) for both wild type control and Akt1s1 null mutants. Each time point was experimentally repeated 2-3 times, generating total 15 samples for the second generation RNA sequencing via Illumina HiSeq platform.