Project description:Testis is the most important male reproductive organ, and the integrity of its physiological function is crucial to the successful production of sperm. In this study, the expression profiles of 11 991 and 8 930 cells in testicular tissue of yak and cattle-yak after sexual maturity were established using Single-cell RNA sequencing. The identification results of cell subpopulations and marker genes were analyzed and their possible mechanisms were predicted.
Project description:Hair follicles of the yak are in anagen and catagen , the hair follicles of healthy female yaks around 2 years old are collected for the preparation of single-cell suspension, the cells were sequenced by scRNA-seq on the 10x genome platform. A total of about 12000 single-cell transcriptome information were obtained. According to the reported marker genes, the main cell groups in anagen and catagen of yak were identified. Based on the analysis of pseudotime trajectory, the differentiation trajectory of epidermal cell lineage and dermal cell lineage during hair follicle development and the dynamic changes of genes during differentiation were described.
Project description:A 1.5-year-old healthy male Hu sheep was selected and castrated for testis collection, then single cell suspension was obtained using enzymatic digestion method for single cell sequencing. And the cell types of sheep testis and marker genes of each cell type were identified based on the RNA sequencing.
Project description:N6-methyladenosine (m6A) is the most prominent mRNA modification in eukaryotes, and its potential regulatory role has recently been identified in mammals, plants, and yeast. However, how m6A methylation regulates spermatogenesis remains unknown. In this study, cattle-yak testis tissue was used as the experimental material, and the m6A map was generated through preliminary experiments and methylated RNA immunoprecipitation sequencing. Only spermatogonia and Sertoli cells were observed in cattle-yak testis tissue. Experiments examining the expression of methylation-related enzymes and the overall methylation level showed that the methylation level in the testis of the cattle-yak was slightly lower than that of the sexually mature yak, but significantly higher than that of the pre-sexually mature yak. Annotation analysis indicated that differentially methylated peaks were most frequently concentrated in exonic regions, followed by 3'UTR and finally 5'UTR regions. Through enrichment analysis of differentially expressed genes and differentially methylated corresponding genes, GO analysis of T-vs-Y group mainly involved spermatogenesis, including cytoskeleton, actin binding, etc. KEGG analysis showed that the differential genes were mainly enriched in actin cytoskeleton regulation and MAPK signaling pathway. GO analysis of the T-vs-M group mainly involved protein ubiquitination, ubiquitin ligase complexes, ubiquitin-dependent protein catabolism and endocytosis. KEGG analysis mainly involved apoptosis and Fanconi anemia pathways. This study will lay the foundation for elucidating the molecular mechanism of m6A in male sterility of cattle-yak.
Project description:A 6-month-old healthy male Hu sheep was selected and castrated for testis collection, then single cell suspension was obtained using enzymatic digestion method for single cell sequencing. And the cell types of sheep testis and marker genes of each cell type were identified based on the RNA sequencing.
Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from P6 ID4-EGFP+ spermatogonia (sorted for brightest or dimmest) or unselected testis cells were used for Drop-Seq analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (a commercial Drop-Seq variant) to perform single-cell RNA-seq