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Microarray analysis to detect whether the genome-wide gene expression profile of fibroblast-induced neurons resembles primary neurons

ABSTRACT: To explore more details in the similarities and differences between ABN+Rarg+Nr5a2 iN cells and primary neurons, we compared the global gene expression pattern of matured iN cells 12 days post infection with mouse primary neurons and MEFs by microarray analysis. In the study presented here, a total of 26423 unique genes were detected in primary mouse embryonic fibroblasts, induced neurons from MEFs by Ascl1+Brn2+Ngn2+Rarg+Nr5a2 at 12 days after infection and primary neurons from postnatal day 0 hippocampus, leading to more details about the similarities and differences among the three groups.

ORGANISM(S): Mus musculus

SUBMITTER: Jianwei Jiao

PROVIDER: E-GEOD-52993 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Microarray analysis to detect whether the genome-wide gene expression profile of fibroblast-induced neurons resembles primary neurons

Project description:To explore more details in the similarities and differences between ABN+Rarg+Nr5a2 iN cells and primary neurons, we compared the global gene expression pattern of matured iN cells 12 days post infection with mouse primary neurons and MEFs by microarray analysis.

2013-12-05 | GSE52993 | GEO

Genome-wide identification and characterization of functional neuronal activity-dependent enhancers

Project description:Experience-dependent gene transcription is required for nervous system development and function. However, the DNA regulatory elements that control this program of gene expression are not well defined. Here we characterize the enhancers that function across the genome to mediate activity-dependent transcription in neurons. While ~12,000 putative activity-regulated enhancer sequences have previously been identified that are enriched for H3K4me1 and the histone acetyltransferase CBP, we find that this chromatin signature is not sufficient to distinguish which of these regulatory sequences are actively engaged in promoting activity-dependent transcription. We show here that a subset of H3K4me1/CBP positive enhancers that is enriched for H3K27 acetylation (H3K27ac) in vivo, and shows increased H3K27ac upon membrane depolarization of cortical neurons, function to regulate activity-dependent transcription. The function of many of these activity-regulated enhancers appears to be dependent on the binding of FOS, a protein that had previously been thought to interact primarily with the promoters of activity-regulated genes. Furthermore, many of these target genes in cortical neurons encode neuron specific proteins that regulate synaptic development and function. These findings suggest that FOS functions at enhancers to control activity-dependent gene programs that are critical for nervous system function, and provide a resource of activity-dependent enhancers that may give insight into genetic variation that contributes to brain development and disease. Genome-wide maps of H3K27ac and AP1 transcription factors (CFOS, FOSB, JUNB) before and after neuronal activity in mouse cortical neurons.

2014-09-07 | E-GEOD-60192 | biostudies-arrayexpress

Transcriptional Modulation of Genes by Gut Microbiota in Germfree and Conventional Mice

Project description:To characterize the effect of microbiota on global gene expression in the distal small intestine during postnatal gut development we employed mouse models with experimental colonization by intestinal microbiota. Using microarray analysis to assess global gene expression in ileal mucosa at the critical stage of intestinal development /maturation associated with weaning, and asking how expression is affected by microbial colonization In the study presented here, preweaned and postweaned GF, SPF mouse small intestinal total RNAs were used. Also, 3-week-old gnotobiotic mouse as well as GF mouse small intestinal RNAs were used.

2013-03-02 | E-GEOD-44788 | biostudies-arrayexpress

Characterization of Human Induced Pluripotent Stem Cell Lines Derived from Neonatal and Adult Human Dermal Fibroblasts

Project description:Somatic cells can be reprogrammed to Induced Pluripotent Stem Cells (iPSCs) by expressing four transcription factors, Oct4, Sox2, Klf4 and c-Myc. Co-expressing Rarg (retinoic acid receptor gamma) and Lrh-1 (liver receptor homolog 1, Nr5a2) with the four factors greatly accelerated reprogramming so that reprogramming of mouse embryonic fibroblast cells (MEFs) to ground state iPSCs requires only four days’ induction of these six factors. The six-factor combination readily reprogrammed primary human neonatal and adult fibroblast cells to exogenous-factor-independent iPSCs, which resembled ground state mouse ES cells in growth properties, gene expression and signalling dependency. Our findings demonstrate that signalling through RARs has critical roles in molecular reprogramming and the synergistic interaction between Rarg and Lrh1 directs reprogramming towards ground state pluripotency. SH-iPS20-1, -3 and -9 lines were derived from HDFa (439656), SH-iPS24-1, -2 and -11 lines were derived from HDFa (709590), and SH-iPS28-23, -25 and -27 lines were derived from HDFn (617769). Parental HDF lines were used as control. SH-iPS15-5 and -10 lines were derived from HDFn (617769, Invitrogen). H1 hESC were obtained from Wicell (Madison, USA). SH-iPSCs were cultured in both 2i+LIF and FGFconditions. Expression pattern of SH-iPSCs was compared to H1 hESC cultured in FGF conditions. S1, S2 and S3 are three indpendent subcultures of a cell line.

2011-10-03 | E-GEOD-32581 | biostudies-arrayexpress

Rat intervertebral disc cell was stimulated by NGF

Project description:The rat-tail intervertbral disc cells was incubated with NGF at a concentration of 100 ng/ml/day for a total of five days. Primary culture of 8 weeks rat-tail interverbral disc cells. The control group was incubated with PBS and the experimental group was incubated with NGF at a concentration of 100 ng/ml/day for a total of five days.

2013-10-01 | E-GEOD-49560 | biostudies-arrayexpress

Neurotoxin tetrodotoxin TTX withdrawal induced neuronal activity: RNA Pol II is poised for rapid induction of arc and other neuronal VF-IEGs

Project description:Transcription of immediate early genes (IEGs) in neurons is exquisitely sensitive to neuronal activity, but the mechanism underlying the earliest of these transcription events is largely unknown. Here we demonstrate that very fast IEGs (VF-IEGs) such as arc/arg3.1 are poised for rapid transcription by the stalling of RNA Polymerase II (Pol II) just downstream of the transcription start site. RNAi-depletion of two subunits of a mediator of Pol II stalling, Negative Elongation Factor, reduces Pol II occupancy of the arc promoter and compromises rapid induction of arc and other VF-IEGs. In contrast, reduction of Pol II stalling did not prevent expression of other fast IEGs (F-IEGs). These F-IEGs are expressed with comparatively slower kinetics and largely lack promoter proximal Pol II stalling. Taken together, our data strongly indicate that very fast kinetics of neuronal IEG expression require poised Pol II and suggest a role for this mechanism in transcription-dependent learning and memory. TTX withdrawal induced neuronal activity. To study activity-induced gene expression, neurons were treated with TTX for 48 hours and then TTX was washed out either for 15 minutes (W15) or for 45 minutes (W45). Gene expression was measured in these two groups in comparison to TTX treated neurons.

2011-04-14 | E-GEOD-22622 | biostudies-arrayexpress

Cav3.2 T-type calcium channel-dependent gene expression after retrieval of context-associated memory (right hippocampus)

Project description:Among all voltage-gated calcium channels, the T-type Ca2+ channels encoded by the Cav3 genes are highly expressed in the hippocampus, which is associated with contextual, temporal and spatial learning and memory. However, the specific involvement of the Cav3.2 T-type Ca2+ channel in these hippocampus-dependent types of learning and memory remains unclear. To investigate the functional role of the 1H channel in learning and memory, we subjected Cav3.2 homozygous, heterozygous knockout and their wild-type littermates to hippocampus-dependent behavioral tasks, including trace fear conditioning (TFC), the Morris water-maze and passive avoidance. The Cav3.2-/- mice performed normally in the Morris water-maze and auditory trace fear conditioning tasks but were impaired in the context-cued trace fear conditioning, step-down and step-through passive avoidance tasks. Furthermore, long-term potentiation (LTP) could be induced for 180 minutes in hippocampal slices of WTs and Cav3.2+/- mice, whereas LTP persisted for only 120 minutes in Cav3.2-/- mice. To determine whether the hippocampal formation is responsible for the impaired behavioral phenotypes , we next performed experiments locally knock down function of the Cav3.2 T-type Ca2+ channel in the hippocampus. Wild-type mice infused with mibefradil exhibited similar behaviors as homozygous knockouts. Finally, microarray analyses indicated that Cav3.2-/- and WT mice presented distinct hippocampal transcriptome profiles. Taken together, our results demonstrate that retrieval of context-associated memory is dependent on the Cav3.2 T-type Ca2+ channel. After WT and Cav3.2 KO mice retrieval of context-associated memory, three right hippocampi of each group were dissected, pooled together and homogenized. The products of experimental and naive groups were used to acquire expression profiles of a total of 29,922 unique genes. Two replicates per group.

2013-02-28 | E-GEOD-30845 | biostudies-arrayexpress

Cav3.2 T-type calcium channel-dependent gene expression after retrieval of context-associated memory

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2013-02-28 | E-GEOD-30846 | biostudies-arrayexpress

Cav3.2 T-type calcium channel-dependent gene expression after retrieval of context-associated memory (left hippocampus)

Project description:Among all voltage-gated calcium channels, the T-type Ca2+ channels encoded by the Cav3 genes are highly expressed in the hippocampus, which is associated with contextual, temporal and spatial learning and memory. However, the specific involvement of the Cav3.2 T-type Ca2+ channel in these hippocampus-dependent types of learning and memory remains unclear. To investigate the functional role of the 1H channel in learning and memory, we subjected Cav3.2 homozygous, heterozygous knockout and their wild-type littermates to hippocampus-dependent behavioral tasks, including trace fear conditioning (TFC), the Morris water-maze and passive avoidance. The Cav3.2-/- mice performed normally in the Morris water-maze and auditory trace fear conditioning tasks but were impaired in the context-cued trace fear conditioning, step-down and step-through passive avoidance tasks. Furthermore, long-term potentiation (LTP) could be induced for 180 minutes in hippocampal slices of WTs and Cav3.2+/- mice, whereas LTP persisted for only 120 minutes in Cav3.2-/- mice. To determine whether the hippocampal formation is responsible for the impaired behavioral phenotypes , we next performed experiments locally knock down function of the Cav3.2 T-type Ca2+ channel in the hippocampus. Wild-type mice infused with mibefradil exhibited similar behaviors as homozygous knockouts. Finally, microarray analyses indicated that Cav3.2-/- and WT mice presented distinct hippocampal transcriptome profiles. Taken together, our results demonstrate that retrieval of context-associated memory is dependent on the Cav3.2 T-type Ca2+ channel. After WT and Cav3.2 KO mice retrieval of context-associated memory, three left hippocampi of each group were dissected, pooled together and homogenized. The products of experimental and naive groups were used to acquire expression profiles of a total of 29,922 unique genes. Two replicates per group.

2013-02-28 | E-GEOD-30842 | biostudies-arrayexpress

Development of distinct gene expression signature in neurons from different regions of the brain

Project description:We have performed microarray expression profiling of mouse primary neurons (cortical neurons and granule cell neurons) to model molecular networks and define whether distinct antiviral IFN responses occurred in neurons corresponding to different brain regions. Primary mouse cortical neurons and granule cell neurons were left untreated, treated with 100IU/mL of IFN-M-NM-2, or infected with West Nile virus (MOI of 1) and RNA was harvested after 24 hours. Three independent experiments were performed using a balanced design.

2012-12-05 | E-GEOD-42727 | biostudies-arrayexpress

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