RESUMO
Early exposure to stressful and adverse life events at fetal and neonatal stages is one of crucial risk factors for mood disorders such as anxiety and depressive disorder in adulthood. Intergenerational effects of prenatal stress on offspring are still not fully understood. We here uncover a significant negative impact of prenatal stress on brain development in embryos and newborns, and on mood-related behaviors and gut microbiota in adult offspring. Prenatal stress leads to reduced numbers in neural progenitors and newborn neurons, and altered gene expression profiles in the mouse embryonic cerebral cortex. Adult mouse offspring exposed to prenatal stress displays altered gene expression in the cortex and elevated responses in anxiety- and depression-like behaviors. Interestingly, prenatal stress has an enduring effect on gut microbiota, as specific microbial community structure is altered in adult F1 offspring treated with prenatal stress, compared to that of the control. Our results highlight the essential impact of prenatal stress on cortical neurogenesis, gene expression patterns, mood-related behaviors, and even gut microbiota in the next generation.
RESUMO
Proper development of the mammalian cerebral cortex relies on precise gene expression regulation. Increasing evidence shows that cortical development is regulated by both mRNAs and long noncoding RNAs (lncRNAs), which also are modified by N6-methyladenosine (m6A). Patterns of m6A-methylation in lncRNAs in the developing cortex have not been uncovered. Here we reveal differentially expressed lncRNAs and report stage-specific m6A-methylation patterns in lncRNAs expressed in mouse embryonic (E) and postnatal (P) cortices using RNA sequencing (RNA-seq) and methylated RNA immunoprecipitation (MeRIP) sequencing. Many lncRNAs show temporal differential expression, and display genic distribution in the genome. Interestingly, we detect temporal-specific m6A-methylation with consensus m6A motif GGACU in the last exon in most lncRNAs. And m6A methylation levels of lncRNAs are positively correlated with the transcript abundance of lncRNAs that have no significantly differential expression in E- and P-stages. Furthermore, the transcript abundance has a positive correlation between the m6A genic lncRNAs and their nearest m6A methylated mRNAs. Our work reveals a fundamental expression reference of lncRNAs and their nearest mRNAs, and highlights an importance of m6A-mediated epitranscriptomic modifications in lncRNAs that are temporally expressed in the developing cortex.
Assuntos
RNA Longo não Codificante , Animais , Córtex Cerebral/metabolismo , Metilação de DNA , Camundongos , RNA Longo não Codificante/metabolismo , RNA Mensageiro , Análise de Sequência de RNARESUMO
N6-methyladenosine (m6A)-mediated epitranscriptomic regulation is critical for various physiological processes. Genetic studies demonstrate that proper m6A-methylation is required for mouse brain development and function. Revealing landscapes of m6A-methylation in the cerebral cortex at different developmental stages will help to understand the biological meaning of epitranscriptomic regulation. Here, we depict the temporal-specific m6A-methylation status in mouse embryonic and postnatal cortices using methylated RNA immunoprecipitation (MeRIP) sequencing. We identified unique m6A binding motifs in stage-specific RNAs and found that more RNA transcripts are temporally methylated in embryonic cortices than in postnatal ones. Moreover, we found that cortical transcription factors and genes associated with neurological disorders are broadly as well specifically methylated at m6A sites. Our study highlights the importance of epitranscriptomic regulation in the developing cortex and provides a fundamental reference for future mechanistic examinations of m6A methylation-mediated gene expression regulation in normal brain development and neurological disorders.
Assuntos
Adenina/análogos & derivados , Córtex Cerebral/metabolismo , Embrião de Mamíferos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Processamento Pós-Transcricional do RNA , RNA/química , Adenina/química , Adenina/metabolismo , Animais , Animais Recém-Nascidos , Córtex Cerebral/citologia , Embrião de Mamíferos/citologia , Perfilação da Expressão Gênica , Metilação , Camundongos , Camundongos Endogâmicos C57BL , RNA/genéticaRESUMO
Proper growth of the mammalian cerebral cortex, which is determined by expansion and survival of neural progenitors and mature neurons, is crucial for cognitive functions. Here, we show a role of the dosage of microRNA miR-9 in controlling brain size. Cortical-specific upregulation of miR-9 causes microcephalic defects in mice, due to apoptosis, reduced neural progenitor pool, and decreased neurogenesis. Glial cell-derived neurotrophic factor (GDNF) is a target of miR-9, and protects neural progenitors from miR-9-induced apoptosis. Furthermore, Zika virus (ZIKV) infection in embryonic mouse cortex causes reduced numbers in neural progenitors and newborn neurons, and results in upregulation of miR-9, downregulation of its target GDNF. Our studies indicate an association of altered levels of miR-9 and its target GDNF with microcephaly and ZIKV infection in mice.
Assuntos
MicroRNAs/genética , Microcefalia/genética , Microcefalia/virologia , Regulação para Cima/genética , Infecção por Zika virus/genética , Animais , Apoptose/genética , Sequência de Bases , Encéfalo/embriologia , Encéfalo/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Camundongos Transgênicos , MicroRNAs/metabolismo , Células-Tronco Neurais/patologia , Células-Tronco Neurais/virologia , Neurogênese/genética , Neurônios/patologia , Infecção por Zika virus/virologiaRESUMO
Long noncoding RNAs (lncRNAs) are emerging as key regulators of crucial cellular processes. However, the molecular mechanisms of many lncRNA functions remain uncharacterized. Sox2ot is an evolutionarily conserved lncRNA that transcriptionally overlaps the pluripotency gene Sox2, which maintains the stemness of embryonic stem cells and tissue-specific stem cells. Here, we show that Sox2ot is expressed in the developing mouse cerebral cortex, where it represses neural progenitor (NP) proliferation and promotes neuronal differentiation. Sox2ot negatively regulates self-renewal of neural stem cells, and is predominately expressed in the nucleus and inhibits Sox2 levels. Sox2ot forms a physical interaction with a multifunctional transcriptional regulator YY1, which binds several CpG islands in the Sox2 locus in a Sox2ot-dependent manner. Similar to Sox2ot, YY1 represses NP expansion in vivo. These results demonstrate a regulatory role of Sox2ot in promoting cortical neurogenesis, possibly by repressing Sox2 expression in NPs, through interacting with YY1.
Assuntos
RNA Longo não Codificante/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Fator de Transcrição YY1/metabolismo , Animais , Diferenciação Celular , Núcleo Celular/metabolismo , Autorrenovação Celular , Córtex Cerebral/metabolismo , Ilhas de CpG , Camundongos , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Neurogênese , Ligação Proteica , Interferência de RNA , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismoRESUMO
The molecular mechanisms regulating neural progenitor (NP) proliferation are fundamental in establishing the cytoarchitecture of the mammalian neocortex. The rate of cell-cycle progression and a fine-tuned balance between cell-cycle re-entry and exit determine the numbers of both NPs and neurons as well as postmitotic neuronal laminar distribution in the cortical wall. Here, we demonstrate that the microRNA (miRNA) miR-210 is required for normal mouse NP cell-cycle progression. Overexpression of miR-210 promotes premature cell-cycle exit and terminal differentiation in NPs, resulting in an increase in early-born postmitotic neurons. Conversely, miR-210 knockdown promotes an increase in the radial glial cell population and delayed differentiation, resulting in an increase in late-born postmitotic neurons. Moreover, the cyclin-dependent kinase CDK7 is regulated by miR-210 and is necessary for normal NP cell-cycle progression. Our findings demonstrate that miRNAs are essential for normal NP proliferation and cell-cycle progress during neocortical development.
Assuntos
Quinases Ciclina-Dependentes/genética , MicroRNAs/genética , Neocórtex/crescimento & desenvolvimento , Neurogênese/genética , Animais , Ciclo Celular/genética , Diferenciação Celular/genética , Proliferação de Células/genética , Embrião de Mamíferos , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Camundongos , Neocórtex/metabolismo , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Quinase Ativadora de Quinase Dependente de CiclinaRESUMO
Methyleugenol (ME) is a natural constituent isolated from many plant essential oils having multiple biological effects including anticonvulsant and anesthetic activities, although the underlying mechanisms remain unclear. Here, we identify ME as a novel agonist of ionotropic γ-aminobutyric acid (GABA) receptors. At lower concentrations (â¼30 µM), ME significantly sensitized GABA-induced, but not glutamate- or glycine-induced, currents in cultured hippocampal neurons, indicative of a preferentially modulatory role of this compound for A type GABA receptors (GABAARs). In addition, ME at higher concentrations (≥100 µM) induced a concentration-dependent, Cl(-)-permeable current in hippocampal neurons, which was inhibited by a GABAAR channel blocker, picrotoxin, and a competitive GABAAR antagonist, bicuculline, but not a specific glycine receptor inhibitor, strychnine. Moreover, ME activated a similar current mediated by recombinant α1-ß2-γ2 or α5-ß2-γ2 GABAARs in human embryonic kidney (HEK) cells. Consequently, ME produced a strong inhibition of synaptically driven neuronal excitation in hippocampal neurons. Together, these results suggest that ME represents a novel agonist of GABAARs, shedding additional light on future development of new therapeutics targeting GABAARs. The present study also adds GABAAR activation to the list of molecular targets of ME that probably account for its biological activities.
Assuntos
Eugenol/análogos & derivados , Agonistas de Receptores de GABA-A/química , Agonistas de Receptores de GABA-A/farmacologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Embrião de Mamíferos , Eugenol/química , Eugenol/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Aminoácidos Excitatórios/farmacologia , GABAérgicos/farmacologia , Ácido Glutâmico/farmacologia , Glicina/farmacologia , Hipocampo/citologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Patch-Clamp , Valina/análogos & derivados , Valina/farmacologiaRESUMO
The embryonic sympathetic nervous system consists of predominantly noradrenergic neurons and a very small population of cholinergic neurons. Postnatal development further allows target-dependent switch of a subset of noradrenergic neurons into cholinergic phenotype. How embryonic cholinergic neurons are specified at the prenatal stages remains largely unknown. In this study, we found that the expression of transcription factor Tlx3 was progressively restricted to a small population of embryonic sympathetic neurons in mice. Immunostaining for vesicular acetylcholine transporter (VAChT) showed that Tlx3 was highly expressed in cholinergic neurons at the late embryonic stage E18.5. Deletion of Tlx3 resulted in the loss of Vacht expression at E18.5 but not E12.5. By contrast, Tlx3 was required for expression of the cholinergic peptide vasoactive intestinal polypeptide (VIP), and somatostatin (SOM) at both E12.5 and E18.5. Furthermore, we found that, at E18.5 these putative cholinergic neurons expressed glial cell line-derived neurotrophic factor family coreceptor Ret but not tyrosine hydroxylase (Ret(+)/TH(-)). Deletion of Tlx3 also resulted in disappearance of high-level Ret expression. Last, unlike Tlx3, Ret was required for the expression of VIP and SOM at E18.5 but not E12.5. Together, these results indicate that transcription factor Tlx3 is required for the acquisition of cholinergic phenotype at the late embryonic stage as well as the expression and maintenance of cholinergic peptides VIP and SOM throughout prenatal development of mouse sympathetic neurons.
Assuntos
Proteínas de Homeodomínio/fisiologia , Neurônios/fisiologia , Neuropeptídeos/fisiologia , Neurotransmissores/fisiologia , Sistema Nervoso Parassimpático/fisiologia , Sistema Nervoso Simpático/fisiologia , Animais , Contagem de Células , Feminino , Feto , Deleção de Genes , Imuno-Histoquímica , Camundongos , Camundongos Knockout , Mutação/fisiologia , Gravidez , Proteínas Proto-Oncogênicas c-ret/biossíntese , Proteínas Proto-Oncogênicas c-ret/genética , Somatostatina/genética , Somatostatina/fisiologia , Gânglio Estrelado/citologia , Gânglio Estrelado/crescimento & desenvolvimento , Sistema Nervoso Simpático/citologia , Sistema Nervoso Simpático/embriologia , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/fisiologia , Peptídeo Intestinal Vasoativo/genética , Peptídeo Intestinal Vasoativo/fisiologia , Proteínas Vesiculares de Transporte de Acetilcolina/genética , Proteínas Vesiculares de Transporte de Acetilcolina/fisiologiaRESUMO
Neural stem cells from different regions within the subventricular zone (SVZ) are able to produce several different subtypes of interneurons in the olfactory bulb throughout life. Previous studies have shown that ischemic stroke induces the production of new neurons in the damaged striatum from the SVZ. However, the origins and genetic profiles of these newborn neurons remain largely unknown as SVZ neural stem cells are heterogeneous. In the present study, using a mouse model of perinatal hypoxic-ischemic (H/I) brain injury combined with BrdU labeling methods, we found that, as in rat brains, virtually all newborn neuroblasts that migrate from the SVZ into the ischemic injured striatum exclusively express the transcription factor Sp8. Furthermore, although newborn neuroblasts are plentiful in the damaged striatum, only a few can differentiate into calretinin-expressing (CR+) interneurons that continuously express Sp8. Genetic fate mapping reveals that newly born CR+ interneurons are generated from Emx1-expressing neural stem cells in the dorsal-lateral SVZ. These results suggest that the fate of the Emx1-expressing lineage in the ischemic damaged striatum is restricted. However, when Sp8 was conditionally inactivated in the Emx1-lineage cells, Pax6 was ectopically expressed by a subpopulation of Emx1-derived CR+ cells in the normal and damaged striatum. Interestingly, these cells possessed large cell bodies and long processes. This work identifies the origin of the newly born CR+ interneurons in the damaged striatum after ischemic brain injury.
Assuntos
Isquemia Encefálica/fisiopatologia , Corpo Estriado/citologia , Interneurônios/fisiologia , Células-Tronco Neurais/fisiologia , Animais , Isquemia Encefálica/patologia , Linhagem da Célula , Corpo Estriado/fisiologia , Proteínas de Ligação a DNA/metabolismo , Proteínas do Domínio Duplacortina , Proteínas do Olho/metabolismo , Proteínas de Homeodomínio/metabolismo , Interneurônios/citologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Associadas aos Microtúbulos/metabolismo , Células-Tronco Neurais/citologia , Neuropeptídeos/metabolismo , Fator de Transcrição PAX6 , Fatores de Transcrição Box Pareados/metabolismo , Ratos , Proteínas Repressoras/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Neuroblasts produced by the neural stem cells of the adult subventricular zone (SVZ) migrate into damaged brain areas after stroke or other brain injuries, and previous data have suggested that they generate regionally appropriate new neurons. To classify the types of neurons produced subsequent to ischemic injury, we combined BrdU or virus labeling with multiple neuronal markers to characterize new cells at different times after the induction of stroke. We show that SVZ neuroblasts give rise almost exclusively to calretinin-expressing cells in the damaged striatum, resulting in the accumulation of these cells during long term recovery after stroke. The vast majority of SVZ neuroblasts as well as newly born young and mature neurons in the damaged striatum constitutively express the transcription factor Sp8, but do not express transcription factors characteristic of medium-sized spiny neurons, the primary striatal projection neurons lost after stroke. Our results suggest that adult neuroblasts do not alter their intrinsic differentiation potential after brain injury.