Project description:<1. The vomeronasal organ (VNO) in the nose of most tetrapods is a sensory structure for the detection of pheromones and kairomones (intra-specific and inter-specific chemical signals) that initiate innate behaviours. It has two neuronal layers, each expressing distinct receptor sub-families coupled to different G-proteins. Neurones in the apical layer express a single vomeronasal receptor type 1 and the G?i2 G-protein. On the other hand, neurons in the basal layer express two or more vomeronasal receptors type-2 (V2Rs) and the G?o G-protein. In mice, V2Rs are organized into four families, A, B, D and C, with family-A and -C sub-divided into subfamilies A1-A10 and C1-C2. These gene families are expressed in a unique combinatorial pattern. Families A (subfamilies A1-A6), B and D are expressed monogenically and coexpress with the recently expanded family-C2 (6 genes, Vmn2r2 to Vmn2r7). While neurons expressing the phylogenetically ancient subfamily-C2 (1 gene, Vmn2r1) coexpress with family-BD and subfamilies A8-A10.We have generated a knock-out mouse for gene Vmn2r1 gene, using a Eucomm tm1b allele. This mouse is a valuable resource for further understanding the molecular organization of the VNO and its behavioural implications, in an evolutionary context. We propose here, to sequence the VNO of Vmn2r1 mutant mice. By studying the VNO transcriptome we hope to advance our knowledge of the genetic coding of pheromonal communication. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:We performed RNaseq of liver, hypothalamus and vomeronasal organ crosses between inbred stains of M. m. domesticus (strain WSB/EiJ) and M. m. musculus (PWD/PhJ). We also sequenced RNA from the same tissues from parental M. m. musculus (PWD/PhJ).
Project description:We applied the NanoString gene expression platform to total RNA isolated from the vomeronasal mucosa of mice of a strain (∆H2Mv) in which we had excised with chromosome engineering a 530 kb genomic region that encompasses nine non-classical class I MHC genes that are expressed specifically in vomeronasal sensory neurons. At three weeks of age, there is no difference of expression for all Vmn2r genes tested; and at eight weeks of age, there is no difference of expression for the majority of Vmn2r genes. We designed NanoString probes for one third of the mouse Vmn2r gene repertoire, based on the reference C57BL/6 genome sequence. CodeSet Chanel includes probes for 46 Vmn2r genes and nine non-classical class I MHC H2-Mv genes that are expressed specifically in mouse vomeronasal sensory neurons. Results were analyzed with tTREAT.
Project description:Our study reports strain specific expression and abensce of sexual dimorphic expression in mouse vomeronasal epithelial tissue Overall design: High throughput sequencing of 32 mice Vomeronasal epithelial tissues with eight samples per strain C57/B6, 129SVEV, SJL, SWR and 4 samples per male and female within each strain
Project description:RNA-Seq has been increasingly used for the quantification and characterization of transcriptomes. The ongoing development of the technology promises the more accurate measurement of gene expression. However, its benefits over widely accepted microarray technologies have not been adequately assessed, especially in toxicogenomics studies. The goal of this study is to enhance the scientific community's understanding of the advantages and challenges of RNA-Seq in the quantification of gene expression by comparing analysis results from RNA-Seq and microarray data on a toxicogenomics study. A typical toxicogenomics study design was used to compare the performance of an RNA-Seq approach (Illumina Genome Analyzer II) to a microarray-based approach (Affymetrix Rat Genome 230 2.0 arrays) for detecting differentially expressed genes (DEGs) in the kidneys of rats treated with aristolochic acid (AA), a carcinogenic and nephrotoxic chemical most notably used for weight loss. We studied the comparability of the RNA-Seq and microarray data in terms of absolute gene expression, gene expression patterns, differentially expressed genes, and biological interpretation. We found that RNA-Seq was more sensitive in detecting genes with low expression levels, while similar gene expression patterns were observed for both platforms. Moreover, although the overlap of the DEGs was only 40-50%, the biological interpretation was largely consistent between the RNA-Seq and microarray data. RNA-Seq maintained a consistent biological interpretation with time-tested microarray platforms while generating more sensitive results. However, there is clearly a need for future investigations to better understand the advantages and limitations of RNA-Seq in toxicogenomics studies and environmental health research. Eight rats were randomly divided into two groups: four rats were administered with aristolochic acid (AA), and four rats were treated with the control vehicle. RNA samples were extracted from the kidney tissue of each rat and were independently assayed with both the NGS (Illumina Genome Analyzer II) and the microarray (Affymetrix Rat Genome 230 2.0) platforms. The RNA-Seq and microarray data were compared in terms of absolute gene expression, gene expression patterns, differentially expressed genes, and biological interpretation.
Project description:The NanoString multiplex gene expression platform was applied to examine changes in V2R gene expression in total RNA samples from the vomeronasal organ of mice of gene-targeted strains ΔC1 and ΔC1-GFP, which each carry a knockout mutation in the Vmn2r1 gene. There is significant decrease in the expression level of various V2R genes that are normally coexpressed with Vmn2r1. Overall design: We designed a custom CodeSet Pao of NanoString probes, which includes probes for 69 V2R genes, covering a half of the mouse V2R gene repertoire, and for nine non-classical class I MHC H2-Mv genes. Total RNA samples from whole vomeronasal mucosae of mice from the ΔC1 and ΔC1-GFP strains were used. Six homozygous mutant mice and six wild-type littermate from each strain were analyzed in one cartridge.
Project description:The mineralocorticoid receptor is expressed in the rat and human retina. We previously showed that intravitreal injection of aldosterone in rat eyes induced retinal œdème and choroidal vasodilation and permeability through regulation of ion/water channels (Zhao et al. Faseb J, 2009; Zhao et al. J Clin Invest 2012). Illicit activation of MR induces inflammation, oxidative stress and tissue remodeling in cardiovascular and renal diseases independent of hypertension. We performed a full transcriptomic study destinated to identify genes regulated by aldosterone in the whole retina of rat.
Project description:Phosphorus, in its orthophosphate form (Pi), is one of the most limiting macronutrients in soils for plant growth and development. However, the whole genome molecular mechanisms contributing to plant acclimation to Pi deficiency remain largely unknown. White lupin (Lupinus albus L.) has evolved unique adaptations for growth in Pi deficient soils including the development of cluster roots to increase root surface area. In this study, we utilized RNA-Seq technology to assess global gene expression in white lupin cluster roots, normal roots, and leaves in response to Pi supply. We de novo assembled 277,224,180 Illumina reads from 12 cDNA libraries to build the first white lupin gene index (LAGI 1.0). This index contains 125,821 unique sequences with an average length of 1,155 bp. Of these sequences 50,734 were transcriptionally active (RPKM = 3) representing approximately 7.8% of the Lupinus albus genome, using the predicted genome size of Lupinus angustifolius as a reference. We identified a total of 2,128 sequences differentially expressed in response to Pi deficiency with a = 2-fold change and a p-value = 0.05. Twelve sequences were consistently differentially expressed due to Pi deficiency stress in three species, making them ideal candidates to monitor the Pi status of plants. Additionally, classic physiological experiments were coupled with RNA-Seq data to examine the role of cytokinin and gibberellic acid in Pi deficiency-induced cluster root development. This global gene expression analysis provides new insights into the biochemical and molecular mechanisms involved in the acclimation to Pi deficiency. Examination of 2 different tissue types (roots and leaves) under phosphorus (P) -sufficient or P-deficient condition with 3 biological replications per condition in white lupin (Lupinus albus).