Identification of transcripts predominantly expressed in cultured chicken primordial germ cells and embryonic fibroblasts.
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ABSTRACT:
ORGANISM(S): Gallus gallus
PROVIDER: GSE38168 | GEO | 2013/01/10
SECONDARY ACCESSION(S): PRJNA167356
REPOSITORIES: GEO
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Project description:The present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between Primordial Germ Cells (PGCs) and their somatic counterpart, chicken embryonic fibroblasts (CEFs). We identified a total of 2,605 expressed transcripts. Among these, 1,197 were predominantly expressed in PGCs, and 1,408 were predominantly expressed in CEFs. Cell culture and microarray data generation were performed in triplicate. Blood PGCs from E2.5 embryos (N=20 for each replication) and CEFs from E6.5 embryos (N=5 for each replication) were cultured in appropriate culture medium. Total RNA was extracted from cultured PGCs (1,500,000 cells for each replication) and cultured CEFs (6,000,000 cells for each replication) with a Qiagen RNeasy kit. About 5 M-BM-5g of total RNA from each replication was used for labeling. Probe synthesis from total RNA samples, hybridization, detection, and scanning were performed according to standard protocols from Affymetrix.
2013-01-10 | E-GEOD-38168 | biostudies-arrayexpress
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:To investigate the function of Lin28A and B in primordial germ cell (PGC) , we generated chicken PGCs in which Lin28A or B has been knocked out by CRISPR/Cas9. We then performed gene expression profiling analysis using data obtained from RNA-seq of 3 different cell types.
2023-05-15 | GSE232032 | GEO
Project description:We report the chromatin status in chicken Embryonic stem cells（ESCs）, Primordial germ cells (PGCs) and Spermatogonial stem cells (SSCs). H3K4me2 antibody was used to collect DNA sequences through CHIP experiments, and the DNA was analyzed by next-generation sequencing. We mapped the genome-wide chromatin maps of chicken ESCs, PGCs, and SSCs. We found that the entire genome of H3K4me2 in ESCs, PGCs, and SSCs was erased and then reconstructed. Most of the enriched genes were related to the cell lineage. It is mainly enriched in distal intergenic. H3K4me2 in the promoter first enriches in ESCs, then decreases in PGCs. The enrichment rebuilds in SSCs. This study provides a basis for analyzing the differences of chromatin status in ESCs, PGCs, and SSCs.
2020-03-21 | GSE147255 | GEO
Project description:Background: Genes, RNAs, and proteins play important roles during germline development. However, the functions of non-coding RNAs (ncRNAs) on germline development remain unclear in avian species. Recent high-throughput techniques have identified several classes of ncRNAs, including micro RNAs (miRNAs), small-interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs). These ncRNAs are functionally important in the genome, however, the identification and annotation of ncRNAs in a genome is challenging. The aim of this study was to identify different types of small ncRNAs particularly piRNAs, and the role of piRNA pathway genes in the protection of chicken primordial germ cells (PGCs). Results: At first, we performed next-generation sequencing to identify ncRNAs in chicken PGCs, and we performed ab initio predictive analysis to identify putative piRNAs in PGCs. Then, we examined the expression of three repetitive sequence-linked piRNAs and 14 genic-transcript-linked piRNAs along with their linked genes using real-time PCR. All piRNAs and their linked genes were highly expressed in PGCs. Subsequently, we knocked down two known piRNA pathway genes of chicken, PIWI-like protein 1 (CIWI) and 2 (CILI), in PGCs using siRNAs. After knockdown of CIWI and CILI, we examined their effects on the expression of six putative piRNA-linked genes and DNA double-strand breakage in PGCs. The knockdown of CIWI and CILI upregulated chicken repetitive 1 (CR1) element and RAP2B, a member of RAS oncogene family, and increased DNA double-strand breakage in PGCs. Conclusions: Our results increase the understanding of PGC-expressed ncRNAs and the role of piRNA pathway genes in the protection of germ cells. Small ncRNA expression profiling in SSEA-1 positive PGCs, SSEA-1 negative gonadal stromal cells (GSCs), Stage X blastoderms, and chicken embryonic fibroblast (CEFs)
2015-04-27 | E-GEOD-60400 | biostudies-arrayexpress
Project description:Background: Genes, RNAs, and proteins play important roles during germline development. However, the functions of non-coding RNAs (ncRNAs) on germline development remain unclear in avian species. Recent high-throughput techniques have identified several classes of ncRNAs, including micro RNAs (miRNAs), small-interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs). These ncRNAs are functionally important in the genome, however, the identification and annotation of ncRNAs in a genome is challenging. The aim of this study was to identify different types of small ncRNAs particularly piRNAs, and the role of piRNA pathway genes in the protection of chicken primordial germ cells (PGCs). Results: At first, we performed next-generation sequencing to identify ncRNAs in chicken PGCs, and we performed ab initio predictive analysis to identify putative piRNAs in PGCs. Then, we examined the expression of three repetitive sequence-linked piRNAs and 14 genic-transcript-linked piRNAs along with their linked genes using real-time PCR. All piRNAs and their linked genes were highly expressed in PGCs. Subsequently, we knocked down two known piRNA pathway genes of chicken, PIWI-like protein 1 (CIWI) and 2 (CILI), in PGCs using siRNAs. After knockdown of CIWI and CILI, we examined their effects on the expression of six putative piRNA-linked genes and DNA double-strand breakage in PGCs. The knockdown of CIWI and CILI upregulated chicken repetitive 1 (CR1) element and RAP2B, a member of RAS oncogene family, and increased DNA double-strand breakage in PGCs. Conclusions: Our results increase the understanding of PGC-expressed ncRNAs and the role of piRNA pathway genes in the protection of germ cells.
2015-04-27 | GSE60400 | GEO
Project description:Generation of genetically uniformed individuals from somatic cells is an effective approach for large-scale reproduction of elite varieties and a powerful tool for restoration of endangered species. However, this technique has never been realized in avian due to their oviparous reproduction pattern. In this study, we produced cloned-like chicken by allogeneic transplantation of somatic cells induced primordial germ cells (PGCs). Oct4, Sox2, Nanog and Lin28A (OSNL) factors were employed to reprogram primary chicken embryo fibroblasts (CEF) to pluripotent stem cells (iPS), in which DNA demethylation and histone acetylation were found to increase the efficiency to 13.00%. The obtained iPS presented embryonic stem cell like characters and were further induced to PGCs by BMP4/BMP8b/EGF, in which histone acetylation and glycolysis inhibition elevated the induction rate to 17.30% and 16.41%, respectively. With the optimized system, we induced Black Langshan Chicken (Gallus domestiaus, black feather) donated CEF to PGCs and transplanted them into the Recessive White Chicken (white feather) embryos. The transplanted cells migrated into the genital ridges and produced functional sperm or oocytes. The sexual matured recipients were self-crossed, with 189/509 (37.13%) cloned-like chicken produced. Microsatellite analysis confirmed their DNA inheritance from the black donor chicken. Thus, we demonstrated, for the first time, the feasibility of avian cloning from somatic cells.
2021-03-16 | GSE159511 | GEO
Project description:The growth factors and signaling pathways that are essential for the inducing proliferation of chicken PGCs is unclear. We investigated the effect of basic fibroblast growth factor (bFGF) on the survival and proliferation of PGCs under feeder-free conditions. We used microarrays to examine the genes regulated by bFGF treatment in chicken PGCs. Cultured PGCs were collected before and after withdrawal of bFGF for 24 h, and 24 h after bFGF replacement. The RNA was extracted and hybridized on Affymetrix microarrays. Three replicates each.
2011-02-01 | E-GEOD-22592 | biostudies-arrayexpress
Project description:The growth factors and signaling pathways that are essential for the inducing proliferation of chicken PGCs is unclear. We investigated the effect of basic fibroblast growth factor (bFGF) on the survival and proliferation of PGCs under feeder-free conditions.
2011-02-01 | GSE22592 | GEO
Project description:The comparative transcriptomic analysis of chicken stem cells defines a chicken set of pluripotency associated genes that is almost coincident with mammalian pluripotency-assocaited genes. A total of 5944 differentially expressed unique identifiers (target IDs) were defined for CEF, 5942 for BM2 cells, 4550 for cES, 5465 for PGC and 5522 for cBC. Chicken Blastodermal cells (cBC) were taken from stage IX-XII (Eyal-Giladi & Kochav, 1976) embryos,chicken embryonic stem (cES) cells were established, amplified on inactivated STO feeder cells in proliferative medium containing cytokines and growth factors as described (Pain et al., 1996, Lavial et al., 2007). Long term cultured primordial germ cells (PGCs) were derived from 48h embryonic blood and maintained as described (McDonald et al., 2010). The non-tumorigenic BM2 monocytic cell line was grown as described (Solari et al., 1996). Primary chicken Embryonic fibroblasts (CEF) were prepared from 11-12 day old embryos according to standard protocols (Gandrillon et al., 1987), maintained and homogenised for 3-4 passages before being used as a somatic cell control. RNAs were extracted using Trizol (Invitrogen) according to the manufacturer and microarray analysis performed in biological triplicate.
2014-11-01 | E-GEOD-61221 | biostudies-arrayexpress