Protein O-GlcNAcylation is a monosaccharide post-translational modification maintained by two evolutionarily conserved enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Mutations in human OGT have recently be...Protein O-GlcNAcylation is a monosaccharide post-translational modification maintained by two evolutionarily conserved enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Mutations in human OGT have recently been associated with neurodevelopmental disorders, although the mechanisms linking O-GlcNAc homeostasis to neurodevelopment are not understood. Here, we investigate the effects of perturbing protein O-GlcNAcylation using transgenic Drosophila lines that overexpress a highly active OGA. We reveal that temporal reduction of protein O-GlcNAcylation in early embryos leads to reduced brain size and olfactory learning in adult Drosophila. Downregulation of O-GlcNAcylation induced by the exogenous OGA activity promotes nuclear foci formation of Polycomb-group protein Polyhomeotic and the accumulation of excess K27 trimethylation of histone H3 (H3K27me3) at the mid-blastula transition. These changes interfere with the zygotic expression of several neurodevelopmental genes, particularly short gastrulation (sog), a component of an evolutionarily conserved sog-Decapentaplegic (Dpp) signaling system required for neuroectoderm specification. Our findings highlight the importance of early embryonic O-GlcNAcylation homeostasis for the fidelity of facultative heterochromatin redeployment and initial cell fate commitment of neuronal lineages, suggesting a possible mechanism underpinning OGT-associated intellectual disability.展开更多
Summary During embryogenesis, plants are thought to use a mechanism that allows the suspensor to maintain its identity. Here, we reported that RPL18a is involved in this mechanism in Arabidopsis thaliana. The suspenso...Summary During embryogenesis, plants are thought to use a mechanism that allows the suspensor to maintain its identity. Here, we reported that RPL18a is involved in this mechanism in Arabidopsis thaliana. The suspensor cells proliferated in rp118aB and formed a multicellular structure rather than undergo pro- grammed cell death, as in wild type. Suspensors of rpl18a expressed the embryo proper marker, DRN:: GFP, but not the suspensor marker, WOX8::GFP. In addition, auxin accumulated throughout the suspensors of rpl18a proembryos. Suspensor-specific expression of RPL18a could rescue the cell proliferation defects in rpl18a suspensors. These findings supported a role for RPL18a in maintaining suspensor identity.展开更多
Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulat...Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulating SE are the microRNAs(miRNAs).Despite previous studies,it is still not clear exactly how miRNAs exert their function of regulating targets during conditionally activated early SE.Here,we use Liriodendron sino-americanum as a model system and perform a combined analysis of microfluidic chips and degradome sequencing to study this process.We identified a total of 386 conserved miRNAs and 153 novel miRNAs during early SE.According to the ANOVA test,239 miRNAs showed 12 distinct expression patterns.Through degradome sequencing,419 targets and 198 targets were identified for 136 known miRNAs and 37 novel miRNAs,respectively.Gene Ontology(GO)and metabolism pathway enrichment analysis revealed that these targets were significantly involved in oxidation-reduction processes,calmodulin-mediated signal transduction pathways and carbohydrate metabolism.The genes that were related to stress responses,phytohormone pathways and plant metabolism were identified within the targets of miR319,miR395,miR408,miR472,miR482,miR390,miR2055,miR156,miR157,miR171,miR396,miR397,miR529,miR535 and miR159.According to promoter analysis,various cis-acting elements related to plant growth and development,phytohormones response and stress response were present in the promoter of the miRNAs.The differential expression patterns of 11 miRNA-target modules were confirmed by real-time quantitative PCR.The study demonstrated that the miRNA plays an important role in the early SE process by regulating its target and then participating in carbohydrate metabolism and stress response.It also provided a valuable resource for further research in determining the genetic mechanism of SE,and then facilitating breeding programs on plants.展开更多
We previously identified a subcortical maternal complex (SCMC) that is essential for early embryogenesis and female fertility in mice. However, the molecular mechanism by which the SCMC affects female fertility rema...We previously identified a subcortical maternal complex (SCMC) that is essential for early embryogenesis and female fertility in mice. However, the molecular mechanism by which the SCMC affects female fertility remains largely uncharacterized. Here, we report that a novel maternal protein, zinc finger BED-type containing 3 (Zbed3), participates in the SCMC. Depletion of maternal Zbed3 results in reduced fecundity of females, because of the impaired and delayed development in a proportion of mutant embryos. The loss of maternal Zbed3 results in asymmetric zygotic division and abnormal distributions of organeUes in the affected oocytes and zygotes, similar to the phenotypes observed in females with disrupted core SCMC genes. Further investiga- tion revealed that these phenotypes are associated with disrupted dynamics of microtubules and/or formation of cytoplasmic lat- tices (CPLs). The stability and localization of Zbed3 depend on, but are not required for, the formation of the SCMC. Thus, our data suggest Zbed3 as one of downstream proteins mediating SCMC functions and provide further insights into the roles of the SCMC and CPLs in female fertility.展开更多
Mammalian embryogenesis begins with a totipotent zygote.Blastocyst-like structures can be captured by aggregated cells with extended pluripotent properties in a three-dimensional(3D)culture system.However,the efficien...Mammalian embryogenesis begins with a totipotent zygote.Blastocyst-like structures can be captured by aggregated cells with extended pluripotent properties in a three-dimensional(3D)culture system.However,the efficiency of generating blastoids is low,and it remains unclear whether other reported totipotent-like stem cells retain a similar capacity.In this study,we demonstrated that spliceosomal repression-induced totipotent blastomere-like cells(TBLCs)form blastocyst-like structures within around 80%of all microwells.In addition,we generated blastoids initiating from a single TBLC.TBLC-blastoids express specific markers of constituent cell lineages of a blastocyst and resemble blastocyst in cell-lineage allocation.Moreover,singlecell RNA sequencing revealed that TBLC-blastoids share a similar transcriptional profile to natural embryos,albeit composed of fewer primitive endoderm-like cells.Furthermore,TBLC-blastoids can develop beyond the implantation stage in vitro and induce decidualization in vivo.In summary,our findings provided an alternative cell type to efficiently generate blastoids for the study of early mouse embryogenesis.展开更多
基金This project has been supported by the National Natural Science Foundation of China(grants 91853108,92153301,31771589,and 32170821 to K.Y,32101034 to F.C)Department of Science and Technology of Hunan Province(grants 2017RS3013,2017XK2011,2018DK2015,2019SK1012,and 2021JJ10054 to K.Y,and the innovative team program 2019RS1010)+2 种基金Central South University(2018CX032 to K.Y,2019zzts046 to Y.Z,2019zzts339 to X.L,2021zzts497 to H.Y,and the innovation-driven team project 2020CX016)D.M.F.v.A.is supported by Wellcome Trust Investigator Award(110061)a Novo Nordisk Foundation Laureate award(NNF21OC0065969).
文摘Protein O-GlcNAcylation is a monosaccharide post-translational modification maintained by two evolutionarily conserved enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Mutations in human OGT have recently been associated with neurodevelopmental disorders, although the mechanisms linking O-GlcNAc homeostasis to neurodevelopment are not understood. Here, we investigate the effects of perturbing protein O-GlcNAcylation using transgenic Drosophila lines that overexpress a highly active OGA. We reveal that temporal reduction of protein O-GlcNAcylation in early embryos leads to reduced brain size and olfactory learning in adult Drosophila. Downregulation of O-GlcNAcylation induced by the exogenous OGA activity promotes nuclear foci formation of Polycomb-group protein Polyhomeotic and the accumulation of excess K27 trimethylation of histone H3 (H3K27me3) at the mid-blastula transition. These changes interfere with the zygotic expression of several neurodevelopmental genes, particularly short gastrulation (sog), a component of an evolutionarily conserved sog-Decapentaplegic (Dpp) signaling system required for neuroectoderm specification. Our findings highlight the importance of early embryonic O-GlcNAcylation homeostasis for the fidelity of facultative heterochromatin redeployment and initial cell fate commitment of neuronal lineages, suggesting a possible mechanism underpinning OGT-associated intellectual disability.
基金supported by the National Natural Science Foundation of China(31570317 and 31270362)
文摘Summary During embryogenesis, plants are thought to use a mechanism that allows the suspensor to maintain its identity. Here, we reported that RPL18a is involved in this mechanism in Arabidopsis thaliana. The suspensor cells proliferated in rp118aB and formed a multicellular structure rather than undergo pro- grammed cell death, as in wild type. Suspensors of rpl18a expressed the embryo proper marker, DRN:: GFP, but not the suspensor marker, WOX8::GFP. In addition, auxin accumulated throughout the suspensors of rpl18a proembryos. Suspensor-specific expression of RPL18a could rescue the cell proliferation defects in rpl18a suspensors. These findings supported a role for RPL18a in maintaining suspensor identity.
基金supported by the Natural Science Foundation of China[32071784]the Qinglan Project of Jiangsu Provincethe Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulating SE are the microRNAs(miRNAs).Despite previous studies,it is still not clear exactly how miRNAs exert their function of regulating targets during conditionally activated early SE.Here,we use Liriodendron sino-americanum as a model system and perform a combined analysis of microfluidic chips and degradome sequencing to study this process.We identified a total of 386 conserved miRNAs and 153 novel miRNAs during early SE.According to the ANOVA test,239 miRNAs showed 12 distinct expression patterns.Through degradome sequencing,419 targets and 198 targets were identified for 136 known miRNAs and 37 novel miRNAs,respectively.Gene Ontology(GO)and metabolism pathway enrichment analysis revealed that these targets were significantly involved in oxidation-reduction processes,calmodulin-mediated signal transduction pathways and carbohydrate metabolism.The genes that were related to stress responses,phytohormone pathways and plant metabolism were identified within the targets of miR319,miR395,miR408,miR472,miR482,miR390,miR2055,miR156,miR157,miR171,miR396,miR397,miR529,miR535 and miR159.According to promoter analysis,various cis-acting elements related to plant growth and development,phytohormones response and stress response were present in the promoter of the miRNAs.The differential expression patterns of 11 miRNA-target modules were confirmed by real-time quantitative PCR.The study demonstrated that the miRNA plays an important role in the early SE process by regulating its target and then participating in carbohydrate metabolism and stress response.It also provided a valuable resource for further research in determining the genetic mechanism of SE,and then facilitating breeding programs on plants.
文摘We previously identified a subcortical maternal complex (SCMC) that is essential for early embryogenesis and female fertility in mice. However, the molecular mechanism by which the SCMC affects female fertility remains largely uncharacterized. Here, we report that a novel maternal protein, zinc finger BED-type containing 3 (Zbed3), participates in the SCMC. Depletion of maternal Zbed3 results in reduced fecundity of females, because of the impaired and delayed development in a proportion of mutant embryos. The loss of maternal Zbed3 results in asymmetric zygotic division and abnormal distributions of organeUes in the affected oocytes and zygotes, similar to the phenotypes observed in females with disrupted core SCMC genes. Further investiga- tion revealed that these phenotypes are associated with disrupted dynamics of microtubules and/or formation of cytoplasmic lat- tices (CPLs). The stability and localization of Zbed3 depend on, but are not required for, the formation of the SCMC. Thus, our data suggest Zbed3 as one of downstream proteins mediating SCMC functions and provide further insights into the roles of the SCMC and CPLs in female fertility.
基金supported by the National Natural Science Foundation of China(32070800)。
文摘Mammalian embryogenesis begins with a totipotent zygote.Blastocyst-like structures can be captured by aggregated cells with extended pluripotent properties in a three-dimensional(3D)culture system.However,the efficiency of generating blastoids is low,and it remains unclear whether other reported totipotent-like stem cells retain a similar capacity.In this study,we demonstrated that spliceosomal repression-induced totipotent blastomere-like cells(TBLCs)form blastocyst-like structures within around 80%of all microwells.In addition,we generated blastoids initiating from a single TBLC.TBLC-blastoids express specific markers of constituent cell lineages of a blastocyst and resemble blastocyst in cell-lineage allocation.Moreover,singlecell RNA sequencing revealed that TBLC-blastoids share a similar transcriptional profile to natural embryos,albeit composed of fewer primitive endoderm-like cells.Furthermore,TBLC-blastoids can develop beyond the implantation stage in vitro and induce decidualization in vivo.In summary,our findings provided an alternative cell type to efficiently generate blastoids for the study of early mouse embryogenesis.