Gene expression profiling analysis of RNA-seq data derived from mice' visual cortex total RNA.
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ABSTRACT: Gene expression profiling analysis of RNA-seq data derived from mice' visual cortex total RNA. Tissues were harvested at specific time points over the day either over a normal 12/12 light/dark cycle, or in a control condition where the mice were also housed in 12/12 conditions but the light did not turn on for the last day, the day of tissue collection.
Project description:RNA-seq libraries purified from the visual cortices of neurons expressing Emx-, GAD2-, PV-, SST-, or VIP-Cre using the Ribotag allele. Seq libraries are provided from mice raised in standard housing, or housed in the dark for two weeks (dark-housed), or dark-housed and then exposed to light for 1, 3, or 7.5 hours. These seq libraries represent the genetic response of distinct types of cortical interneurons to altered sensory experience.
Project description:Visual deprivation, either in the form of dark rearing (DR) or monocular deprivation (MD) are established paradigms for studying cortical plasticity. We have used miRNA microarray to uncover miRNAs whose expression is altered in primary visual cortex following DR and/or MD. C57BL6 mice were reared in normal light and dark conditions (control) till P28, in complete darkness since birth (DR) till P28, or were grown in normal light/dark conditions from birth till P24 and then subjected to lid suturing of one eye till P28. Mice were euthanized at P28 and their primary visual cortex areas were excised and subjected to RNA isolation. In the case of MD mice only the contralateral to lid suture primary visual cortex was extracted. 100ng of total RNA (tested and quantified using the Agilent Bioanalyzer 2100) were labeled using the Agilent miRNA labeling system and hybridized to Agilent murine miRNA arrays. Microarrays were hybridized overnight at 64 ºC, scanned using an Agilent scanner and extracted with Agilent feature extractor 10.1.
Project description:RNA-seq libraries purified from the visual cortices of neurons expressing Emx-, GAD2-, PV-, SST-, or VIP-Cre using the Ribotag allele. Seq libraries are provided from mice raised in standard housing, or housed in the dark for two weeks (dark-housed), or dark-housed and then exposed to light for 1, 3, or 7.5 hours. These seq libraries represent the genetic response of distinct types of cortical interneurons to altered sensory experience. To explore how sensory experience affects gene expression, we examined this process in the visual cortex of adult mice that were housed in standard conditions, in complete darkness (i.e. dark-housed), or dark-housed and then exposed to light for increasing amounts of time. We generated mice that were heterozygous for alleles of either Emx-,Gad2-,Sst-,Vip- or Pv-Cre, and were also heterozygous for the Rpl22-HA (RiboTag) allele, which expresses an HA-tagged ribosomal protein specifically in Cre-expressing neurons. We performed RNA-Seq on RNA isolated from the dark-housed/light-exposed RiboTag-mice; Experiments were done in 3 biological replicates and the visual cortices of 3 mice were pooled per sample at each time-point and for each Cre line.
Project description:Analysis of thalamus and hypothalamus under conditions of visual deprivation by dark-rearing (DR). Animals subjected to DR from birth till postnatal day (P) 14. Results provide insight into the role of visual inputs in the regulation of gene expression in thalamus and hypothalamus during development.
Project description:Wild-type (C57Bl6) mice were dark reared from birth to P42, then underwent monocular enucleation at P42. They were then placed in a normal light-dark environment for 4 days, after which they were sacrificed (at P46). Visual cortex was dissected from 1-mm thick coronal sections, and total RNA was extracted from the tissue lysate using Trizol (Gibco-BRL). Keywords: repeat
Project description:Analysis of thalamus and hypothalamus under conditions of visual deprivation by dark-rearing (DR). Animals subjected to DR from birth till postnatal day (P) 14. Results provide insight into the role of visual inputs in the regulation of gene expression in thalamus and hypothalamus during development. RNA sample was taken from thalamus and hypothalamus of 14-day-old mouse reared in standard or dark condition. Comparisons among groups were made by one-color method with normalized data from Cy3 channels for data analysis.
Project description:Rats were housed in either long day (LD, 16:8 hours, light:dark) or short day (SD, 8:16 hours, light dark) photoperiods for 3, 14 and 28 days and brains removed, frozen on dry ice and then hypothalamic arcuate nucleus tissue blocks cut for RNA extraction. At each time point 4 rats were used in each photoperiod giving 24 rats in total.
Project description:Visual system development is light-experience dependent, which strongly implicates epigenetic mechanisms in light-regulated maturation. Among many epigenetic processes, genomic imprinting is an epigenetic mechanism through which monoallelic gene expression occurs in a parent-of-origin-specific manner. It is unknown if genomic imprinting contributes to visual system development. We profiled the transcriptome and imprintome during critical periods of mouse visual system development under normal- and dark-rearing conditions using B6/CAST F1 hybrid mice. We identified experience-regulated, isoform-specific, and brain region-specific imprinted genes. We also found imprinted microRNAs were predominantly clustered into the Dlk1-Dio3 imprinted locus with light experience affecting some imprinted miRNA expression. Our findings provide the first comprehensive analysis of light-experience regulation of the transcriptome and imprintome during critical periods of visual system development. Our results may contribute to therapeutic strategies for visual impairments and circadian rhythm disorders resulting from a dysfunctional imprintome.
Project description:Visual deprivation, either in the form of dark rearing (DR) or monocular deprivation (MD) are established paradigms for studying cortical plasticity. We have used miRNA microarray to uncover miRNAs whose expression is altered in primary visual cortex following DR and/or MD.
Project description:Wild-type (C57Bl6) mice were dark reared from birth to P42, then underwent monocular enucleation at P42. They were then placed in a normal light-dark environment for 4 days, after which they were sacrificed (at P46). Visual cortex was dissected from 1-mm thick coronal sections, and total RNA was extracted from the tissue lysate using Trizol (Gibco-BRL). Experiment Overall Design: Gene expression in pooled samples from 2 non-deprived (ipsilateral to monocular enucleation) and 2 deprived (contralateral to monocular enucleation) visual hemispheres was compared. Note: two additional cross-animal comparisons were made.