Single-cell RNA sequencing(scRNA-seq)is useful for exploring cell heterogeneity.For large animals,however,little is known regarding spermatogonial stem cell(SSC)selfrenewal regulation,especially in dairy goats.In this...Single-cell RNA sequencing(scRNA-seq)is useful for exploring cell heterogeneity.For large animals,however,little is known regarding spermatogonial stem cell(SSC)selfrenewal regulation,especially in dairy goats.In this study,we described a high-resolution scRNA-seq atlas derived from a dairy goat.We identified six somatic cell and five spermatogenic cell subtypes.During spermatogenesis,genes with significantly changed expression were mainly enriched in the Notch,TGF-β,and Hippo signaling pathways as well as the signaling pathway involved in the regulation of stem cell pluripotency.We detected and screened specific candidate marker genes(TKTL1 and AES)for spermatogonia.Our study provides new insights into goat spermatogenesis and the development of testicular somatic cells.展开更多
Double sex and mab-3-related transcription factor 1(Dmrt1),which is expressed in goat male germline stem cells(mGSCs)and Sertoli cells,is one of the most conserved transcription factors involved in sex determination.I...Double sex and mab-3-related transcription factor 1(Dmrt1),which is expressed in goat male germline stem cells(mGSCs)and Sertoli cells,is one of the most conserved transcription factors involved in sex determination.In this study,we highlighted the role of Dmrt1 in balancing the innate immune response in goat mGSCs.Dmrt1 recruited promyelocytic leukemia zinc finger(Plzf),also known as zinc finger and BTB domain-containing protein 16(Zbtb16),to repress the Toll-like receptor 4(TLR4)-dependent inflammatory signaling pathway and nuclear factor(NF)-κB.Knockdown of Dmrt1 in seminiferous tubules resulted in widespread degeneration of germ and somatic cells,while the expression of proinflammatory factors were significantly enhanced.We also demonstrated that Dmrt1 stimulated proliferation of mGSCs,but repressed apoptosis caused by the immune response.Thus,Dmrt1 is sufficient to reduce inflammation in the testes,thereby establishing the stability of spermatogenesis and the testicular microenvironment.展开更多
基金supported by the National Natural Science Foundation of China(31572399,32072806,32072815,32002246,61772431,62072377)Program of Shaanxi Province Science and Technology Innovation Team(2019TD-036)+4 种基金Program of State Key Lab of Reproductive Regulation&Breeding of Grassland Livestock(SKL-OT-201801)Program of State Key Laboratory of Respiratory Disease(SKLRD-OP-202114)Science and Technology Major Project of Inner Mongolia Autonomous Region of China(ZDZX2018065)the First-class University and Academic Program from Northwest A&F University(Z1010221003)Financial aid for basic operation fee of Central University(Z1090219146)。
文摘Single-cell RNA sequencing(scRNA-seq)is useful for exploring cell heterogeneity.For large animals,however,little is known regarding spermatogonial stem cell(SSC)selfrenewal regulation,especially in dairy goats.In this study,we described a high-resolution scRNA-seq atlas derived from a dairy goat.We identified six somatic cell and five spermatogenic cell subtypes.During spermatogenesis,genes with significantly changed expression were mainly enriched in the Notch,TGF-β,and Hippo signaling pathways as well as the signaling pathway involved in the regulation of stem cell pluripotency.We detected and screened specific candidate marker genes(TKTL1 and AES)for spermatogonia.Our study provides new insights into goat spermatogenesis and the development of testicular somatic cells.
基金This work was supported by the China National Basic Research Program(2016YFA0100203)National Natural Science Foundation of China(31572399Detail,32072806,32072815,32002246)+3 种基金State Key Lab of Reproductive Regulation&Breeding of Grassland Livestock(SKL-OT-201801)Science and Technology Major Project of Inner Mongolia Autonomous Region of China(ZDZX2018065)and Shaanxi Province Science and Technology Innovation Team(2019TD-036)The authors thank Dr.John Clotaire Daguia Zambe for helpful comments about this paper,Jia Fang for the PGL3-NF-κB luciferase reporter plasmid,and Dong-Xue Che for bioinformatics analysis.
文摘Double sex and mab-3-related transcription factor 1(Dmrt1),which is expressed in goat male germline stem cells(mGSCs)and Sertoli cells,is one of the most conserved transcription factors involved in sex determination.In this study,we highlighted the role of Dmrt1 in balancing the innate immune response in goat mGSCs.Dmrt1 recruited promyelocytic leukemia zinc finger(Plzf),also known as zinc finger and BTB domain-containing protein 16(Zbtb16),to repress the Toll-like receptor 4(TLR4)-dependent inflammatory signaling pathway and nuclear factor(NF)-κB.Knockdown of Dmrt1 in seminiferous tubules resulted in widespread degeneration of germ and somatic cells,while the expression of proinflammatory factors were significantly enhanced.We also demonstrated that Dmrt1 stimulated proliferation of mGSCs,but repressed apoptosis caused by the immune response.Thus,Dmrt1 is sufficient to reduce inflammation in the testes,thereby establishing the stability of spermatogenesis and the testicular microenvironment.
