Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:A transcriptome study in mouse hematopoietic stem cells was performed using a sensitive SAGE method, in an attempt to detect medium and low abundant transcripts expressed in these cells. Among a total of 31,380 unique transcript, 17,326 (55%) known genes were detected, 14,054 (45%) low-copy transcripts that have no matches to currently known genes. 3,899 (23%) were alternatively spliced transcripts of the known genes and 3,754 (22%) represent anti-sense transcripts from known genes.
Project description:A transcriptome study in mouse hematopoietic stem cells was performed using a sensitive SAGE method, in an attempt to detect medium and low abundant transcripts expressed in these cells. Among a total of 31,380 unique transcript, 17,326 (55%) known genes were detected, 14,054 (45%) low-copy transcripts that have no matches to currently known genes. 3,899 (23%) were alternatively spliced transcripts of the known genes and 3,754 (22%) represent anti-sense transcripts from known genes. Mouse hematopoietic stem cells were purified from bone marrow cells using negative and positive selection with a Magnetic-Activated Cell Sorter (MACS). total RNA and mRNA were purified from the purified cells using Trizol reagent and magnetic oligo dT beads. Double strand cDNAs were synthesized using a cDNA synthesis kit and anchored oligo dT primers. After NlaIII digestion, 3’ cDNAs were isolated and amplified through 16-cycle PCR. SAGE tags were released from the 3’ cDNA after linker ligation. Ditags were formed, concatemerized and cloned into a pZERO vector. Sequencing reactions were performed with the ET sequencing terminator kit. Sequences were collected using a Megabase 1000 sequencer. SAGE tag sequences were extracted using SAGE 2000 software.
Project description:C57BL6 mice (12-15 weeks old) were obtained from Charles River Laboratories (St. Constant, Québec). They were housed in an air-conditioned room (19-25Åé) with controlled lighting from 07:15 to 19:15 h and were given free access to laboratory chow (Lab Rodent Diet No. 5002, Ren’s Feed and Suppliers, Ontario) and water. The hypothalamus, pituitary gland and parietal cortex of 51 male and 51 randomly estrous cycling female mice were pooled and used for expression profiling. After vertebral cervical dislocation under isoflurane anesthesia, the brain and pituitary gland were removed from the skull and the three organs were immediately dissected, snap-frozen in liquid nitrogen and stored at -80Åé until further analysis. Keywords: other
Project description:The regulation of pituitary function via the hypothalamus and via intra-pituitary connections represents a complex system. Though hormones secreted from the pituitary glands have been well studied, overall information of proteins expressed in the pituitary glands is very limited. Protein expression profiling of normal pituitary tissue may lead to discovery of novel proteins playing an important role in the physiology of pituitary glands and can lead to better understanding of pituitary gland diseases. We aimed to carry out systematic proteomic profiling of adenohypophysis from human pituitary glands using high-resolution Fourier transform mass spectrometer. A total of 2,175 proteins were identified in this study of which, 105 proteins were identified for the first time as compared to high throughput proteomic-based studies from human pituitary glands. The comprehensive list of proteins identified in this study will facilitate the better understanding the role of this important gland in health and disease.
Project description:Purpose: The goal of this study was to compare gene expression patterns in the male and female human cortex Methods: We performed RiboZero Gold (rRNA depleted) 50bp PE RNA-seq in a set of control samples of both sexes to identify sexually dimorphic gene expression patterns. Results: Within these samples, we corroborated findings from a discovery set of RNA-seq data from adult human cortex tissue from the BrainSpan consortium which demonstrated male-biased expression of astrocyte marker genes and a gene co-expression module found to be up-regulated in the adult autistic cortex. Conclusions: These findings suggest that sex-differential risk for autism spectrum disorder is not the result of sex-differential regulation of ASD risk genes, but of naturally occurring sexually dimorphic processes that modulate the impact of risk variants for autism spectrum disorder. 13 cerebral cortex samples from 10 individuals (7 samples from 5 males, 6 samples from 5 females). Three Samples are included in this study from Series GSE64018. **PLEASE NOTE: Raw data has been submitted to dbGAP**
Project description:C57BL6 mice (12-15 weeks old) were obtained from Charles River Laboratories (St. Constant, Québec). They were housed in an air-conditioned room (19-25Åé) with controlled lighting from 07:15 to 19:15 h and were given free access to laboratory chow (Lab Rodent Diet No. 5002, Ren’s Feed and Suppliers, Ontario) and water. The hypothalamus, pituitary gland and parietal cortex of 51 male and 51 randomly estrous cycling female mice were pooled and used for expression profiling. After vertebral cervical dislocation under isoflurane anesthesia, the brain and pituitary gland were removed from the skull and the three organs were immediately dissected, snap-frozen in liquid nitrogen and stored at -80Åé until further analysis. Keywords: other
Project description:Introgressed variants from other species can be an important source of genetic variation because they may arise rapidly, can include multiple mutations on a single haplotype, and have often been pretested by selection in the species of origin. Although introgressed alleles are generally deleterious, several studies have reported introgression as the source of adaptive alleles-including the rodenticide-resistant variant of Vkorc1 that introgressed from Mus spretus into European populations of Mus musculus domesticus. Here, we conducted bidirectional genome scans to characterize introgressed regions into one wild population of M. spretus from Spain and three wild populations of M. m. domesticus from France, Germany, and Iran. Despite the fact that these species show considerable intrinsic postzygotic reproductive isolation, introgression was observed in all individuals, including in the M. musculus reference genome (GRCm38). Mus spretus individuals had a greater proportion of introgression compared with M. m. domesticus, and within M. m. domesticus, the proportion of introgression decreased with geographic distance from the area of sympatry. Introgression was observed on all autosomes for both species, but not on the X-chromosome in M. m. domesticus, consistent with known X-linked hybrid sterility and inviability genes that have been mapped to the M. spretus X-chromosome. Tract lengths were generally short with a few outliers of up to 2.7 Mb. Interestingly, the longest introgressed tracts were in olfactory receptor regions, and introgressed tracts were significantly enriched for olfactory receptor genes in both species, suggesting that introgression may be a source of functional novelty even between species with high barriers to gene flow.