Single-cell RNA sequencing reveals atlas of dairy goat testis cells

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.

Dmrt1 regulates the immune response by repressing the TLR4 signaling pathway in goat male germline stem cells

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.

LIN28A inhibits DUSP family phosphatases and activates MAPK signaling pathway to maintain pluripotency in porcine induced pluripotent stem cells

LIN28A,an RNA-binding protein,plays an important role in porcine induced pluripotent stem cells(piPSCs).However,the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs remains unclear.Here,we explored the function of LIN28A in piPSCs based on its overexpression and knockdown.We performed total RNA sequencing(RNA-seq)of piPSCs and detected the expression levels of relevant genes by quantitative real-time polymerase chain reaction(qRT-PCR),western blot analysis,and immunofluorescence staining.Results indicated that piPSC proliferation ability decreased following LIN28A knockdown.Furthermore,when LIN28A expression in the shLIN28A2 group was lower(by 20%)than that in the negative control knockdown group(shNC),the pluripotency of piPSCs disappeared and they differentiated into neuroectoderm cells.Results also showed that LIN28A overexpression inhibited the expression of DUSP(dual-specificity phosphatases)family phosphatases and activated the mitogen-activated protein kinase(MAPK)signaling pathway.Thus,LIN28A appears to activate the MAPK signaling pathway to maintain the pluripotency and proliferation ability of piPSCs.Our study provides a new resource for exploring the functions of LIN28A in piPSCs.