Expression of putative spermatogonial markers DDX6, NAP1L4 and TKTL1 in the pre-pubertal, pubertal and post-pubertal bovine testis (#222)
Spermatogonial stem cells (SSCs) offer potential applications in transgenic breeding, reproductive applications, and our general understanding of stem cell biology. However, few advances have been made toward fully characterising and understanding the biology of bovine spermatogonial stem cells and their development. Protein markers associated with germ cells in bovine testicular tissue can be used to develop our understanding of how germ cells, and in particular spermatogonial stem cells, develop over the life span of the bovine. Recently, we identified a series of potential marker candidates for bovine germ cell in a proteomic analysis of bovine testis cells combined with transcriptional analysis. Here we show expression patterns of the potential testis cell markers, DDX6, NAP1L4 and TKTL1, in pre-pubertal, pubertal and adult bovine testis tissue, in comparison to ZBTB16 (PLZF), a marker for undifferentiated spermatogonia, DDX4 (VASA), a marker of the germ cell lineage and vimentin, a Sertoli cell marker. In pre-pubertal tissue, none of the protein markers show co-staining with vimentin, indicating they are not expressed by Sertoli cells. DDX6 was associated with both early and later stage spermatogonia, including cells stained by DBA-biotin. NAP1L4 showed strong overlap for DBA-biotin staining; indicating its association with early stage spermatogonia. TKTL1 showed staining of cells towards the centre of the seminiferous tubules consistent with expression profiles of advanced spermatogonia. Staining testis tissue of Bos Taurus bulls at different developmental stages, pre pubertal, peri pubertal and post pubertal (adult), provides an insight into the development of the bovine testis, with particular interest in how spermatogonia develop and are arranged within the testis throughout the life of the animal.