MicroRNAs are small non-coding RNAs that play crucial roles in the regulation of gene expression and protein synthesis during brain development. MiR-3099 is highly expressed throughout embryogenesis, especially in the...MicroRNAs are small non-coding RNAs that play crucial roles in the regulation of gene expression and protein synthesis during brain development. MiR-3099 is highly expressed throughout embryogenesis, especially in the developing central nervous system. Moreover, miR-3099 is also expressed at a higher level in differentiating neurons in vitro, suggesting that it is a potential regulator during neuronal cell development. This study aimed to predict the target genes of miR-3099 via in-silico analysis using four independent prediction algorithms（miRDB,miRanda, Target Scan, and DIANA-micro-T-CDS） with emphasis on target genes related to brain development and function. Based on the analysis, a total of 3,174 miR-3099 target genes were predicted. Those predicted by at least three algorithms（324 genes） were subjected to DAVID bioinformatics analysis to understand their overallfunctional themes and representation. The analysis revealed that nearly 70% of the target genes were expressed in the nervous system and a significant proportion were associated with transcriptional regulation and protein ubiquitination mechanisms. Comparison of in situ hybridization（ISH） expression patterns of miR-3099 in both published and in-house-generated ISH sections with the ISH sections of target genes from the Allen Brain Atlas identified 7 target genes（Dnmt3a, Gabpa, Gfap, Itga4,Lxn, Smad7, and Tbx18） having expression patterns complementary to miR-3099 in the developing and adult mouse brain samples. Of these, we validated Gfap as a direct downstream target of miR-3099 using the luciferase reporter gene system. In conclusion, we report the successful prediction and validation of Gfap as an miR-3099 target gene using a combination of bioinformatics resources with enrichment of annotations based on functional ontologies and a spatio-temporal expression dataset.展开更多
In the mammalian central nervous system,nerve-glia antigen 2(NG2)glia are considered the fourth glial population in addition to astrocytes,oligodendrocytes and microglia.The fate of NG2 glia in vivo has been carefully...In the mammalian central nervous system,nerve-glia antigen 2(NG2)glia are considered the fourth glial population in addition to astrocytes,oligodendrocytes and microglia.The fate of NG2 glia in vivo has been carefully studied in several transgenic mouse models using the Cre/loxP strategy.There is a clear agreement that NG2 glia mainly serve as progenitors for oligodendrocytes and a subpopulation of astrocytes mainly in the ventral forebrain,whereas the existence of a neurogenic potential of NG2 glia is lack of adequate evidence.This mini review summarizes the findings from recent studies regarding the fate of NG2 glia during development.We will highlight the age-and-region-dependent heterogeneity of the NG2 glia differentiation potential.We will also discuss putative reasons for inconsistent findings in various transgenic mouse lines of previous studies.展开更多
Astroglia are integral to brain development and the emergence of neurodevelopmental disorders.However,studying the pathophysiology of human astroglia using brain organoid models has been hindered by ineficient astrogl...Astroglia are integral to brain development and the emergence of neurodevelopmental disorders.However,studying the pathophysiology of human astroglia using brain organoid models has been hindered by ineficient astrogliogene-sis.In this study,we introduce a robust method for generating astroglia-enriched organoids through BMP4 treatment during the neural differentiation phase of organoid development.Our RNA sequencing analysis reveals that astroglia developed within these organoids exhibit advanced developmental characteristics and enhanced synaptic functions compared to those grown under traditional two-dimensional conditions,particularly highlighted by increased neu-rexin(NRXN)-neuroligin(NLGN)signaling.Cell adhesion molecules,such as NRXN and NLGN,are essential in regulat-ing interactions between astroglia and neurons.We further discovered that brain organoids derived from human embryonic stem cells(hESCs)harboring the autism-associated NLGN3 R451C mutation exhibit increased astroglio-genesis.Notably,the NLGN3 R451C astroglia demonstrate enhanced branching,indicating a more intricate morphol-ogy.Interestingly,our RNA sequencing data suggest that these mutant astroglia significantly upregulate pathways that support neural functions when compared to isogenic wild-type astroglia.Our findings establish a novel astroglia-enriched organoid model,offering a valuable platform for probing the roles of human astroglia in brain development and related disorders.展开更多
Background Neural stem cells （NSCs） are a self-renewing and multipotent population of the central nervous system （CNS）, which are active during development and maintain homeostasis and tissue integrity throughout ...Background Neural stem cells （NSCs） are a self-renewing and multipotent population of the central nervous system （CNS）, which are active during development and maintain homeostasis and tissue integrity throughout life. Microglias are an immune cell population resident in the CNS, which have crucial physiological functions in the developing and adult CNS. This study aimed to investigate that whether microglia co-cultured with NSCs could promote astrogliogenesis from NSCs. Methods Microglia and NSCs were co-cultured in 24-well insert plates. NSCs were plated in the bottom of the well and microglia in the insert. Fluorescent staining, Western blotting and RT-PCR were used to determine the effect of microglia on NSCs differentiation. Results Co-culture of microglia and NSCs promoted astrogliogenesis from NSCs. Several key genes, such as Notch 1, Notch 2, Notch 3, Hes 5, and NRSFwere downregulated, while the critical genes Idl and Id2 were upregulated. BMP2 and FGF2 were upregulated. Conclusion Microglias act as a regulator of NSCs astrogliogenesis.展开更多
基金supported by the Science Fund(02-01-04-SF2336)the Fundamental Research Grant Scheme,Ministry of Higher Education,Malaysia(FRGS-04-01-15-1663FR)
文摘MicroRNAs are small non-coding RNAs that play crucial roles in the regulation of gene expression and protein synthesis during brain development. MiR-3099 is highly expressed throughout embryogenesis, especially in the developing central nervous system. Moreover, miR-3099 is also expressed at a higher level in differentiating neurons in vitro, suggesting that it is a potential regulator during neuronal cell development. This study aimed to predict the target genes of miR-3099 via in-silico analysis using four independent prediction algorithms（miRDB,miRanda, Target Scan, and DIANA-micro-T-CDS） with emphasis on target genes related to brain development and function. Based on the analysis, a total of 3,174 miR-3099 target genes were predicted. Those predicted by at least three algorithms（324 genes） were subjected to DAVID bioinformatics analysis to understand their overallfunctional themes and representation. The analysis revealed that nearly 70% of the target genes were expressed in the nervous system and a significant proportion were associated with transcriptional regulation and protein ubiquitination mechanisms. Comparison of in situ hybridization（ISH） expression patterns of miR-3099 in both published and in-house-generated ISH sections with the ISH sections of target genes from the Allen Brain Atlas identified 7 target genes（Dnmt3a, Gabpa, Gfap, Itga4,Lxn, Smad7, and Tbx18） having expression patterns complementary to miR-3099 in the developing and adult mouse brain samples. Of these, we validated Gfap as a direct downstream target of miR-3099 using the luciferase reporter gene system. In conclusion, we report the successful prediction and validation of Gfap as an miR-3099 target gene using a combination of bioinformatics resources with enrichment of annotations based on functional ontologies and a spatio-temporal expression dataset.
基金This work was supported by grants from the Deutsche Forschungsgemeinschaft DFG Sino-German joint project(Kl 503/14-1)to WH,DFG FOR 2289 to ASfrom the Saarland University Medical Faculty HOMFOR2015 and HOMFORexzellenz2016 to AS and WH,respectivelyWH was also supported by DFG SFB 894 and the European Commission EC-H2020 FET ProAct Neurofibres.
文摘In the mammalian central nervous system,nerve-glia antigen 2(NG2)glia are considered the fourth glial population in addition to astrocytes,oligodendrocytes and microglia.The fate of NG2 glia in vivo has been carefully studied in several transgenic mouse models using the Cre/loxP strategy.There is a clear agreement that NG2 glia mainly serve as progenitors for oligodendrocytes and a subpopulation of astrocytes mainly in the ventral forebrain,whereas the existence of a neurogenic potential of NG2 glia is lack of adequate evidence.This mini review summarizes the findings from recent studies regarding the fate of NG2 glia during development.We will highlight the age-and-region-dependent heterogeneity of the NG2 glia differentiation potential.We will also discuss putative reasons for inconsistent findings in various transgenic mouse lines of previous studies.
基金supported by grants from the NIH(R01NS102382,R01NS122108,and R01AG073779 to P.J.)M.J.was supported by a post-doctoral fellowship award from the New Jersey Department of Health(CAUT24DFP004)+1 种基金A.V.P.was supported by a graduate trainee T32 fellowship award from the Training in Translating Neuroscience to Therapies program at Rutgers University(T32NS115700)L.C.was supported by the Rutgers HealthAdvance Fund(NHLBI U01HL150852 to L.C.)。
文摘Astroglia are integral to brain development and the emergence of neurodevelopmental disorders.However,studying the pathophysiology of human astroglia using brain organoid models has been hindered by ineficient astrogliogene-sis.In this study,we introduce a robust method for generating astroglia-enriched organoids through BMP4 treatment during the neural differentiation phase of organoid development.Our RNA sequencing analysis reveals that astroglia developed within these organoids exhibit advanced developmental characteristics and enhanced synaptic functions compared to those grown under traditional two-dimensional conditions,particularly highlighted by increased neu-rexin(NRXN)-neuroligin(NLGN)signaling.Cell adhesion molecules,such as NRXN and NLGN,are essential in regulat-ing interactions between astroglia and neurons.We further discovered that brain organoids derived from human embryonic stem cells(hESCs)harboring the autism-associated NLGN3 R451C mutation exhibit increased astroglio-genesis.Notably,the NLGN3 R451C astroglia demonstrate enhanced branching,indicating a more intricate morphol-ogy.Interestingly,our RNA sequencing data suggest that these mutant astroglia significantly upregulate pathways that support neural functions when compared to isogenic wild-type astroglia.Our findings establish a novel astroglia-enriched organoid model,offering a valuable platform for probing the roles of human astroglia in brain development and related disorders.
文摘Background Neural stem cells （NSCs） are a self-renewing and multipotent population of the central nervous system （CNS）, which are active during development and maintain homeostasis and tissue integrity throughout life. Microglias are an immune cell population resident in the CNS, which have crucial physiological functions in the developing and adult CNS. This study aimed to investigate that whether microglia co-cultured with NSCs could promote astrogliogenesis from NSCs. Methods Microglia and NSCs were co-cultured in 24-well insert plates. NSCs were plated in the bottom of the well and microglia in the insert. Fluorescent staining, Western blotting and RT-PCR were used to determine the effect of microglia on NSCs differentiation. Results Co-culture of microglia and NSCs promoted astrogliogenesis from NSCs. Several key genes, such as Notch 1, Notch 2, Notch 3, Hes 5, and NRSFwere downregulated, while the critical genes Idl and Id2 were upregulated. BMP2 and FGF2 were upregulated. Conclusion Microglias act as a regulator of NSCs astrogliogenesis.
