Project description:Purpose: Study the differentially expressed genes in hippocampal rat CA1 regions 90 minutes after Long-Term Potentiation (LTP) was induced using field recordings Methods: hippocampal rat CA1 region mRNA profiles of control (unstimulated) and 90 minutes after LTPwas induced (LTP 90) were generated by deep sequencing. A pool of approximately 10 CA1 regions collected from at least 3 different animals were used. The seq library was prepared using TruSeq RNA Sample Prep Kit v2 protocol (Illumina, CA, USA). For the RNA-Seq data analysis Tophat 2.0.13, bowtie 2.1.0 , samtool 0.1.7 and cufflinks 1.3.0 were used. The rn5-bowtie2 index was generated with the command 'bowtie2-build rn5.fa rn5'. The 'rn5.fa'-file was downloaded from the UCSC genome browser. The mm10-bowtie2 index was downloaded from http://bowtie-bio.sourceforge.net/bowtie2/manual.shtml. RefSeq geneTracks and GTF-files for the rn5 and mm10 genome assembly were downloaded from UCSC genome browser. Common gene ids in the GTF-files were matched to individual transcript_ids using the corresponding official symbols obtained from the geneTracks files. Conclusions: Our study illustrates the nature of the different transcripts in acute rat hippocampal slices after LTP 90 minutes was induced using field electrophysiology and compares them with control, unstimulated slices. Thius allowed us to identify wich genes are modulated in the hippocampus in order to promote and sustain LTP.
Project description:Transcriptional profiling of 3dpf wild type zebrafish larvae treated with 20mM PTZ for 30 and 90 minutes compared with 3dpf wild type control untreated zebrafish larvae.
Project description:Goal of the study was to investigate genes regulated differentially after 90 minutes versus 7 days after irradiation. 7 day time point corresponds to cellular senescence. U2OS cells were irradiated with 20 Gy and harvested either 90 minutes or 7 days. In addition cells were left untreated (control) or had knockdown of NFKBIA (protein name Ikappa B alpha). Ikappa B alpha is the main inhibitor of the transcription factor NFkappa B. NFkappa B regulates transcription of many factors constituting the SASP (senescence associated secretory phenotype).
Project description:Parathyroid hormone (PTH) plays an essential role in regulating calcium and bone homeostasis in the adult, but whether PTH is required at all for regulating fetal-placental mineral homeostasis is uncertain. To address this we treated Pth-null mice in utero with 1 nmol PTH (1-84) or saline and examined placental calcium transfer 90 minutes later. It was found that placental calcium transfer increased in Pth-null fetuses treated with PTH as compared to Pth-null fetuses treated with saline. Subsequently, to determine the effect of PTH treatment on placental gene expression, in a separate experiment, 90 minutes after the fetal injections the placentas were removed for subsequent RNA extraction and microarray analysis.
Project description:This study was aimed at elucidating the mechanisms underlying activity-dependent gene regulation during long-term potentiation (LTP) of mouse hippocampal CA3-CA1 synapses. Transcriptome analysis allowed to identify changes in gene expression and alternative splicing induced 1 hour and 3 hours after LTP induction.
Project description:In this study we used microarray analysis to reveal the gene expression profile of the hippocampal CA1 subregion, which was laser-capture microdissected one week after kainic acid (KA)-induced status epilepticus (SE) in postnatal day 21 (P21) rats. These rats are developmentally roughly comparable to juvenile children, and KA-induced SE leads to selective damage of hippocampal CA1 pyramidal neurons in this age group while saving neurons of the other sub-regions. We searched for alterations in the gene expression pattern during the early epileptogenetic phase, i.e. one week after SE, and compared the results with those of age-matched control rats. To detect specifically changes in the CA1 pyramidal neurons, we used the laser-capture microdissection technique that allows the precise isolation of the region of interest. The RNA of this region was isolated, amplified, and labeled, and then hybridized to Illumina RatRef-12 Expression BeadChip Arrays. The gene expression data generated from the microarray was first normalized by the guantile normalization method, and then filtered by using the empirical Bayes method, and the contrasts were created by using the Limma R/Bioconductor. Finally, the data was clustered by using the non-hierarchical K-means clustering for genes, and the pathway analysis was performed by âGene set testâ, which analyzes the statistical significance of a set of genes simultaneously ranked by p-value and generates the KEGG categories (Chipster manual). The Illumina microarray analysis with the Chipster software v1.1.0 (http://chipster.csc.fi; CSC, Espoo, Finland) generated a total of 1592 differently expressed genes in the CA1 subregion of KA-treated rats compared to control rats. Using the K-means method the genes were classified in 10 different clusters. The subsequent KEGG-test for the probe set over-representation analysis revealed the 15 significantly (p<0.05) changed KEGG-pathways in response to KA-treatment, e.g. oxidative phosphorylation (26 genes changed), and long-term potentiation (LTP; 18 genes changed). Some of the differentially expressed genes were also identified to be involved in Ca2+ homeostasis, gliosis, inflammation, and GABAergic transmission.
Project description:Polysome-associated transcriptome analysis in cells after 90 minutes of mistranslation induction comparing with a pool of mRNAs existent in cells without induction at several time points (Reference)
Project description:We performed ATAC-seq in the GR18 cell line (derived from U2OS ATTC:HTB-96, stably transfected with an expression construct for GR), upon glucocorticoid (dexamethasone) or vehicle (ethanol) treatment for 90 minutes.