MPSS profiling of embryonic gonad and primordial germ cells
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ABSTRACT:
ORGANISM(S): Gallus gallus
PROVIDER: GSE5909 | GEO | 2007/03/01
SECONDARY ACCESSION(S): PRJNA97345
REPOSITORIES: GEO
Similar Datasets
Project description:We used the MPSS technology to uncover gene expression profiling in a greater depth in the early embryonic gonads and primordial germ cells (PGCs) in the chicken. Total numbers of sequenced signatures were 1,012,533 and 995,676 for the PGCs and gonad. Using a false discovery rate cut-off of 0.05, we found 3.05 % of all signatures in the PCGs and 1.89 % of all signatures in the gonad signatures were significantly up-regulated compared to each sample. The MPSS result was very consistent with our previous result using EST data(http://chickgce.snu.ac.kr/). Keywords: cell type comparison Experimental animals provided for this experiment were maintained at the University Animal Farm, Seoul National University, and all experimental procedures were performed at the affiliated laboratories of the university. Gonadal cells were retrieved from the gonads of 6.5-day-old (stage 29) White Leghorn (WL) embryos by our standard procedure [26]. Embryos were freed from the yolk by rinsing with calcium- and magnesium-free PBS and the gonads were retrieved by dissection of embryo abdomen with sharp tweezers under a stereomicroscope. Embryonic gonads were collected from total 1,947 embryos by 10 highly-skilled persons through 8 separated experimental batches. Gonadal tissues were dissociated by gentle pipetting in 0.05% (v:v) trypsin solution supplemented with 0.53 mM EDTA. After centrifuged at 200xg for 5 min, total gonadal cells were loaded into MACS (Miltenyi Biotech, Germany), and the separated primordial germ cells (PGCs) were immediately stored in liquid nitrogen (-190ºC) until processed further. The numbers of PGCs in cell population before and after loading were counted. MACS treatment for chicken PGCs and counting PGC number Chicken gonadal cells were incubated with PGC-specific primary antibody, anti-stage specific embryo antigen (anti-SSEA)-1 antibody for chicken PGCs (mouse IgM isotype), for 20 min at the room temperature of 20-25?C. Anti-SSEA-1 antibody developed by [27] was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Development of Biological Science. After washing with 1mL buffer (PBS supplement with 0.5% BSA and 2mM EDTA), the supernatant was completely removed. The pellet was mixed with 100 ? buffer containing 20? of rat anti-mouse IgM microbeads for 15 min at 4?C. Treated cells were carefully washed by the addition of 500 ? buffer and subsequently loaded with MACS.[28] For counting cell number, chicken PGCs before or after MACS treatment were fixed with 1% (v:v) glutaraldehyde for 5 min and rinsed with 1x PBS twice. The anti-SSEA-1 ascites fluid diluted 1:1,000 in PBS was added and subsequent steps were carried out using DAKO universal LSAB® kit, Peroxidase (DAKO, USA) according to the manufacturer’s instruction. After 8 batches of cell preparation, total cell number of PGC-enriched fraction and gonadal stromal cells was 5.26X106 and 1.76X108, respectively. These cell populations were further used for total RNA isolation and massively parallel signature sequencing (MPSS) analysis.
2007-02-28 | E-GEOD-5909 | biostudies-arrayexpress
Project description:MPSS mouse transcriptome analysis project. See http://www.ncbi.nlm.nih.gov/geo/info/mouse-trans.html for more details. Keywords: other
2004-10-26 | GSE1581 | GEO
Project description:Primordial germ cells (PGCs), the embryonic precursors of eggs and sperm, are a unique model for identifying and studying regulatory mechanisms in singly migrating cells. From their time of specification to eventual colonization of the gonad, mouse PGCs traverse through and interact with many different cell types, including epithelial cells and mesenchymal tissues. Work in drosophila and zebrafish have identified many genes and signaling pathways involved in PGC migration, but little is known about this process in mammals. We have generated a point mutation in the Ror2 gene that we know disrupts primordial germ cell migration in the developing mouse embryo. We used microarray analysis to determine if this defect is mediated through genome-wide or pathway-specific transcriptional changes. We analyzed primordial germ cells (PGCs) from 4 wild-type (WT) and 4 Ror2Y324C/Y324C mutant embryos using Oct4-DPE-EGFP. PGCs were collected during their active migratory state at embryonic day 9.5 (somite range 20-25).
2014-08-08 | E-GEOD-60179 | biostudies-arrayexpress
Project description:This series represents a very deep survey of all transcripts expressed in a wide range of human tissues and cells by Massively Parallel Signature Sequencing (MPSS). These datasets have been generated and donated to the scientific community by Lynx Therapeutics, Inc in Hayward, CA. The data contained in this submission is described in the following publication: 'C. Victor Jongeneel, Mauro Delorenzi, Christian Iseli, Daixing Zhou, Christian D. Haudenschild, Brian J. Stevenson, Robert L. Strausberg, Andrew J.G. Simpson, and Thomas J. Vasicek.' An atlas of human gene expression from massively parallel signature sequencing (MPSS). Genome Research VolXX 2004 Abstract of the publication: 'We have used massively parallel signature sequencing (MPSS) to sample the transcriptomes of 32 normal human tissues to an unprecedented depth, thus documenting the patterns of expression of over 20,000 genes with high sensitivity and specificity. The data confirm the widely held belief that differences between cell and tissue types are largely determined by the expression of a limited number of tissue-specific genes, rather than by combinations of more promiscuously expressed genes. Expression of a little over half of all known human genes seems to account for both the common requirements and the specific functions of the tissues sampled. A classification of tissues based on patterns of gene expression largely reproduces classifications based on anatomical and biochemical properties. The unbiased sampling of the human transcriptome achieved by MPSS supports the notion that most human genes have been mapped, if not functionally characterised. This dataset should prove useful for the identification of tissue-specific genes, for the study of global changes induced by pathological conditions, and for the definition of a minimal set of genes necessary for basic cell maintenance.' Keywords: other
2005-11-29 | GSE1747 | GEO
Project description:A comparison of microarray and MPSS technologies can help to establish the metrics for data comparisons across these technology platforms and determine some of the factors affecting the measurement of mRNA abundances using different platforms. Here, different Treatments/Conditions based on different Arabidopsis tissues were used for three different platforms include MPSS, Affymetrix and Agilent. Keywords: MPSS, Affymetrix, Agilent
2007-09-14 | GSE8994 | GEO
Project description:In poultry, in vitro derived primordial germ cells (PGCs) represent an important tool for management of genetic resources. However, several studies have highlighted sexual differences exhibited by PGCs through in vitro steps, which may compromise their reproductive capacities. To understand this phenomenon, we compared the proteome of pregonadal chicken male (ZZ) and female (ZW) PGCs expanded in vitro by quantitative proteomic analysis using a GeLC-MS/MS strategy. The proteins found to be differentially abundant in chicken male and female PGCs indicated their early sexual identity. Many of the proteins up-accumulated in male PGCs were encoded by genes strongly enriched in the sexual chromosome Z. This suggests that the known lack of dosage compensation of the transcription of Z-linked genes between sexes persists at protein level in PGCs, and that this may be a key factor of their autonomous sex differentiation. Male and female PGCs up-accumulated protein sets were associated with differential biological processes, and contained proteins biologically relevant for male and female germ cell development respectively. This study presents first evidence on early predetermined sex specific cell fate of chicken PGCs that will help to understand their sexual physiological specificities and enable more precise sex-specific adaptation of in vitro culture conditions.
2021-11-03 | PXD022452 | Pride
Project description:SAGE and MPSS libraries were produced from the same RNA sample extracted from an activated CD4+ T cell clone in order to compare the ability of these techniques to indentify the full range of genes expressed in a single cell type. Keywords: Technical comparison of tag-based technologies SAGE and MPSS
2007-08-02 | GSE8612 | GEO
Project description:MPSS analysis of mouse BLK CL.4 cells and liver cells. Total RNA, nuclear RNA and post-nuclear RNA examined. Keywords: other
2004-10-26 | GSE1067 | GEO
Project description:We report the use of MPSS data to compare transcripts expressed in wild-type and sporocyteless mutant ovules All MPSS libraries were performed as described by Meyers et al (Nature Biotechnol. 2004 Aug;22(8):1006-11. Epub 2004 Jul 11.)
2013-08-21 | E-GEOD-50020 | biostudies-arrayexpress
Project description:Primordial germ cells (PGCs), the embryonic precursors of eggs and sperm, are a unique model for identifying and studying regulatory mechanisms in singly migrating cells. From their time of specification to eventual colonization of the gonad, mouse PGCs traverse through and interact with many different cell types, including epithelial cells and mesenchymal tissues. Work in drosophila and zebrafish have identified many genes and signaling pathways involved in PGC migration, but little is known about this process in mammals. We have generated a point mutation in the Ror2 gene that we know disrupts primordial germ cell migration in the developing mouse embryo. We used microarray analysis to determine if this defect is mediated through genome-wide or pathway-specific transcriptional changes.
2014-08-08 | GSE60179 | GEO