Project description:The maintenance of spermatogenesis in adult males is dependent on a population of mitotic germ cells with self-renewal potential known as undifferentiated spermatogonia. Regulation of undifferentiated spermatogonia function is dependent on transcriptional and post-transcriptional mechanisms. We have identified an essential role for the RNA helicase DDX5 in undifferentiated spermatogonia through generation of a UBC-CreERT2;Ddx5flox/flox mouse model that allows tamoxifen-dependent Ddx5 ablation. To identify genes regulated by DDX5, we generated lines of cultured undifferentiated spermatogonia from these mice and treated cells with tamoxifen (TMX) to induce Ddx5 knockout or vehicle (VEH) as a control, and performed RNA-sequencing analysis to compare these conditions.
Project description:The undifferentiated spermatogonial population of mouse testis is functionally heterogeneous and contains stem cells and committed progenitor cells. However, gene expression patterns marking these distinct cell fractions are poorly defined. Therefore, our aim was to profile gene expression of undifferentiated cells by RNA-Seq at the single cell level. Undifferentiated cells were therefore isolated from adult mouse testes based on expression of a Plzf/Zbtb16 gene reporter, which marks both stem and progenitor spermatogonia. Isolated cells were then processed using the 10X Chromium system and analysed by RNA-Seq. Our goal was to characterize distinct subsets of undifferentiated spermatogonia based on gene expression patterns.
Project description:The undifferentiated spermatogonial population of mouse testis is known to be functionally heterogeneous and contain both stem cells and committed progenitor cells. However, gene expression patterns marking these distinct cell fractions are poorly defined. We found that a subset of undifferentiated spermatogonia were marked by expression of an Oct4-GFP transgene but properties of these cells were unclear. Undifferentiated cells were therefore isolated from adult testes and seperated according to expression of Oct4-GFP for gene expression analysis by microarray. Our goal was to correlate Oct4-GFP expression with that of known markers of stem and committed progenitor cells to determine which undifferentiated cell populations were marked by Oct4-GFP.