Similar Datasets
Project description:Neural stem cells (NSCs) have astroglial hallmarks, while astrocytes in the intact adult mammalian brain parenchyma are thought to be postmitotic and lack NSC hallmarks that are activated only after injury. Analysis of genome-wide expression at population and single cell level in astrocytes from the diencephalon (DIE) and the cerebral cortex Grey grey Matter matter (CTX GM) revealed cell cycle associated gene expression, that we verified in situ by PCNA immunostaining and incorporation of the thymidine analog EdU. Strikingly, clonal analysis using GLASTCreERT2/confetti mice showed that only diencephalic astrocytes give rise to daughter cells in multi-cellular clones. In addition, subsets of astrocytes only from the diencephalon form multipotent, self-renewing neurospheres in vitro. Given the differential expression levels of genes related to Smad mediated transcriptional regulation between CTX GM and DIE astrocytes, we deleted the common mediator Smad4 in adult astrocytes. This abrogates proliferation of diencephalic astrocytes in vivo and their neurosphere formation in vitro. This work therefore identified a novel astrocyte subtype with NSC hallmarks in the diencephalon and implicates ongoing astrocytogenesis as a region-specific hallmark in the adult brain.
2021-09-01 | GSE149112 | GEO
Project description:Neural stem cells (NSCs) have astroglial hallmarks, while astrocytes in the intact adult mammalian brain parenchyma are thought to be postmitotic and lack NSC hallmarks that are activated only after injury. Analysis of genome-wide expression at population and single cell level in astrocytes from the diencephalon (DIE) and the cerebral cortex Grey grey Matter matter (CTX GM) revealed cell cycle associated gene expression, that we verified in situ by PCNA immunostaining and incorporation of the thymidine analog EdU. Strikingly, clonal analysis using GLASTCreERT2/confetti mice showed that only diencephalic astrocytes give rise to daughter cells in multi-cellular clones. In addition, subsets of astrocytes only from the diencephalon form multipotent, self-renewing neurospheres in vitro. Given the differential expression levels of genes related to Smad mediated transcriptional regulation between CTX GM and DIE astrocytes, we deleted the common mediator Smad4 in adult astrocytes. This abrogates proliferation of diencephalic astrocytes in vivo and their neurosphere formation in vitro. This work therefore identified a novel astrocyte subtype with NSC hallmarks in the diencephalon and implicates ongoing astrocytogenesis as a region-specific hallmark in the adult brain.
2021-09-01 | GSE149114 | GEO
Project description:Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent
| PRJNA627405 | ENA
Project description:Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent – RNAseq
| PRJNA627403 | ENA
Project description:Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent – scRNA-seq
| PRJNA627410 | ENA
Project description:Astrocytes regulate brain-wide functions and also show region-specific differences, but little is known about how general and region-specific functions are aligned at the single-cell level. To explore this, we isolated adult mouse diencephalic astrocytes by ACSA-2-mediated magnetic-activated cell sorting (MACS). Single-cell RNA-seq revealed 7 gene expression clusters of astrocytes, with 4 forming a supercluster. Within the supercluster, cells differed by gene expression related to ion homeostasis or metabolism, with the former sharing gene expression with other regions and the latter being restricted to specific regions. All clusters showed expression of proliferation-related genes, and proliferation of diencephalic astrocytes was confirmed by immunostaining. Clonal analysis demonstrated low level of astrogenesis in the adult diencephalon, but not in cerebral cortex grey matter. This led to the identification of Smad4 as a key regulator of diencephalic astrocyte in vivo proliferation and in vitro neurosphere formation. Thus, astrocytes show diverse gene expression states related to distinct functions with some subsets being more widespread while others are more regionally restricted. However, all share low-level proliferation revealing the novel concept of adult astrogenesis in the diencephalon.
| S-EPMC8561644 | biostudies-literature
Project description:Astrocytes regulate brain-wide functions and also show region-specific differences, but little is known about how general and region-specific functions are aligned at the single cell level. To explore this, we isolated adult mouse diencephalic astrocytes by ACSA2-mediated magnetic-activated cell sorting (MACS). Single cell RNA-seq revealed 7 gene expression clusters of astrocytes, with 4 forming a super cluster. Within the super cluster, cells differed by gene expression related to ion homeostasis or metabolism, with the former sharing gene expression with other regions and the latter being restricted to specific regions. All clusters showed expression of proliferation related genes, and proliferation of diencephalic astrocytes was confirmed by immunostaining. Clonal analysis demonstrated low level of astrogenesis in the adult diencephalon, but not in cortex Grey Matter. This led to the identification of Smad4 as a key regulator of diencephalic astrocyte in vivo proliferation and in vitro neurosphere formation. Thus, astrocytes show diverse gene expression states related to distinct functions with some subsets being more wide-spread while others are more regionally restricted. However, all share low level proliferation revealing the novel concept of adult astrogenesis in the diencephalon.
| S-SCDT-EMBOJ-2020-107532 | biostudies-other
Project description:In eukaryotes, histone variant distribution within the genome is the key epigenetic feature. To understand how each histone variant is targeted to the genome, we developed a new method, the RhIP (Reconstituted histone complex Incorporation into chromatin of Permeabilized cell) assay, in which epitope-tagged histone complexes are introduced into permeabilized cells and incorporated into their chromatin. Using this method, we found that H3.1 and H3.3 were incorporated into chromatin in replication-dependent and -independent manners, respectively. We further found that the incorporation of histones H2A and H2A.Z mainly occurred at less condensed chromatin (open), suggesting that condensed chromatin (closed) is a barrier for histone incorporation. To overcome this barrier, H2A, but not H2A.Z, uses a replication-coupled deposition mechanism. Our study revealed that the combination of chromatin structure and DNA replication dictates the differential histone deposition to maintain the epigenetic chromatin states.
| S-EPMC8110306 | biostudies-literature
Project description:BackgroundBipolar disorder is a highly heritable neuropsychiatric condition affecting more than 1% of the human population. Lithium salts are commonly prescribed as a mood stabilizer for individuals with bipolar disorder. Lithium is clinically effective in approximately half of treated individuals, and their genetic backgrounds are known to influence treatment outcomes. While the mechanism of lithium's therapeutic action is unclear, it stimulates adult neural progenitor cell proliferation, similar to some antidepressant drugs.MethodsTo identify common genetic variants that modulate lithium-induced proliferation, we conducted an EdU incorporation assay in a library of 80 genotyped human neural progenitor cells treated with lithium. These data were used to perform a genome-wide association study to identify common genetic variants that influence lithium-induced neural progenitor cell proliferation. We manipulated the expression of a putatively causal gene using CRISPRi/a (clustered regularly interspaced short palindromic repeats interference/activation) constructs to experimentally verify lithium-induced proliferation effects.ResultsWe identified a locus on chr3p21.1 associated with lithium-induced proliferation. This locus is also associated with bipolar disorder risk, schizophrenia risk, and interindividual differences in intelligence. We identified a single gene, GNL3, whose expression temporally increased in an allele-specific fashion following lithium treatment. Experimentally increasing the expression of GNL3 led to increased proliferation under baseline conditions, while experimentally decreasing GNL3 expression suppressed lithium-induced proliferation.ConclusionsOur experiments reveal that common genetic variation modulates lithium-induced neural progenitor proliferation and that GNL3 expression is necessary for the full proliferation-stimulating effects of lithium. These results suggest that performing genome-wide associations in genetically diverse human cell lines is a useful approach to discover context-specific pharmacogenomic effects.
| S-EPMC9982734 | biostudies-literature
Project description:Animals bear communities of gut microorganisms with substantial effects on animal nutrition, but the host genetic basis of these effects is unknown. Here we use Drosophila to demonstrate substantial among-genotype variation in the effects of eliminating the gut microbiota on five host nutritional indices (weight, protein, lipid, glucose and glycogen contents); this includes variation in both the magnitude and direction of microbiota-dependent effects. Genome-wide association studies to identify the genetic basis of the microbiota-dependent variation reveal polymorphisms in largely non-overlapping sets of genes associated with variation in the nutritional traits, including strong representation of conserved genes functioning in signalling. Key genes identified by the GWA study are validated by loss-of-function mutations that altered microbiota-dependent nutritional effects. We conclude that the microbiota interacts with the animal at multiple points in the signalling and regulatory networks that determine animal nutrition. These interactions with the microbiota are probably conserved across animals, including humans.
| S-EPMC4333721 | biostudies-literature