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Differential expression of glycosyltransferases in P19 mouse embryonal carcinoma cells with or without differentiation by retinoic acid

ABSTRACT: The project is aimed at understanding the role of the Lex-containing L5 carbohydrate epitope in the developmental process of neural induction. We plan to identify 1) the protein to which the L5 glycan is attached, 2) glycosyltransferases upregulated during neural induction, 3) the corresponding L5 carbohydrate binding protein (CBP), 4) the mechanism of interaction between L5 and its CBP, and 5) the spatiotemporal regulation of L5 and L5-CBP expression during neural induction. There is evidence, by Westernblot and immunofluorescence, that the expression of the Lewis-x-like L5 carbohydrate epitope shows dramatic differences in expression between undifferentiated cells and cells differentiated with retinoic acid. We wish to know which enzyme(s) involved in the biosynthetic pathway may be up/downregulated under these experimental conditions. Two classes of p19 mouse embryonal carcinoma cell RNA were analyzed: undifferentiated and differentiated with retinotic acid. Classes were prepared in triplicate for a total of 6 samples. All samples were hybridized to the custom designed CFG GLYCOv2 glycogene array.

ORGANISM(S): Mus musculus

SUBMITTER: Steven Head

PROVIDER: E-GEOD-27596 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Differential expression of glycosyltransferases in P19 mouse embryonal carcinoma cells with or without differentiation by retinoic acid

Project description:The project is aimed at understanding the role of the Lex-containing L5 carbohydrate epitope in the developmental process of neural induction. We plan to identify 1) the protein to which the L5 glycan is attached, 2) glycosyltransferases upregulated during neural induction, 3) the corresponding L5 carbohydrate binding protein (CBP), 4) the mechanism of interaction between L5 and its CBP, and 5) the spatiotemporal regulation of L5 and L5-CBP expression during neural induction. There is evidence, by Westernblot and immunofluorescence, that the expression of the Lewis-x-like L5 carbohydrate epitope shows dramatic differences in expression between undifferentiated cells and cells differentiated with retinoic acid. We wish to know which enzyme(s) involved in the biosynthetic pathway may be up/downregulated under these experimental conditions.

2011-03-02 | GSE27596 | GEO

Expression data from Oct4+ cell fraction in SFEBq cultured mouse embryonic stem cells

Project description:To further characterize residual undifferentiated cells after neural induction of embryonic stem cells, we performed DNA microarray analysis to identify genes expressed predominantly in residual undifferentiated cells expressing Oct4. We used microarrays to detail the global programme of gene expression underlying cellularization and identified distinct classes of up-regulated genes in residual undifferentiated cells after neural induction of mouse embryonic stem cells by SFEBq.

2016-11-17 | GSE89912 | GEO

Technical comparison of qRT-PCR experiments with the GLYCOv2 custom array

Project description:Gene expression patterns were surveyed using RNA from 4 different murine sources. Four classes of RNA were analyzed: undifferentiated embryonic stem cells, differentiated ES cells, kidney and liver tissues. Classes were prepared in triplicate for a total of 12 samples. All samples were hybridized to the custom designed CFG GLYCOv2 glycogene array. Analysis results were used for comparison with parallel experiments using qRT-PCR.

2011-03-02 | E-GEOD-27595 | biostudies-arrayexpress

Interleukin-22 (IL-22) regulates B3GNT7 expression to induce intestinal fucosylation of O-linked glycans

Project description:Interleukin (IL) - 22 is a cytokine that plays a critical role in the maintenance of intestinal epithelial homeostasis. Its downstream functions are mediated through ligation of its heterodimeric receptor comprised of IL-22 receptor subunit alpha 1 (IL-22Ra1) and IL-10R2, and subsequent activation of signal transducer and activator of transcription 3 (STAT3). IL-22 signaling can induce genes necessary for intestinal epithelial cell proliferation, tissue regeneration, tight junction fortification, and production of antimicrobials. Emerging studies have also implicated IL-22 signaling in the regulation of intestinal epithelial fucosylation in mice. However, whether IL-22 regulates intestinal fucosylation in human intestinal epithelial cells and the molecular mechanisms that govern this process are unknown. Here, we show that IL-22 signaling regulates expression of the B3GNT7 transcript, which encodes a β1-3-N-acetylglucosaminyltransferase that can participate in the synthesis of poly-N-acetyllactosamine (LacNAc) chains. Additionally, we show that IL-22 signaling regulates levels of the a1-3-fucosylated Lewis X (Lex) blood group antigen and this glycan epitope is primarily displayed on heavily O-glycosylated intestinal epithelial glycoproteins. The IL-22-mediated increase in Lex antigen levels requires B3GNT7, as disruption of B3GNT7 expression moderated the IL-22-dependent increase in Lex antigen levels. Moreover, we show that increased expression of B3GNT7 alone is sufficient to promote increased display of Lex-decorated carbohydrate glycan structures primarily on O-glycosylated intestinal epithelial glycoproteins. Together, the data presented here identify B3GNT7 as a critical intermediary in IL-22-dependent induction of intestinal fucosylation of O-linked glycans and uncover a novel role for B3GNT7 in intestinal glycosylation.

2021-11-27 | GSE154889 | GEO

Glycogene expression changes between different passages in the 4T1 Mouse metastatic breast cancer cell line

Project description:This laboratory's focus is on the immune response to carbohydrate antigens, with an emphasis on modalities to analyze and to increase the immune response. We are also interested in carbohydrate drug targeting. The hypothesis is that these 2 variants have different cell surface carbohydrate expression, which results in different adhesion patterns, which ultimately results in different metastasis patterns. Evidence for expression differences and loss of the variants ability to metastasize to the lung could have clinical implications in the prevention of metastasis to different organs. The 4T1 mouse metastatic breast cancer cell line is known to change with passages in culture. Our data has shown changes cause the cells to metastasize to the intestine rather than to the lung. Four classes of RNA were analyzed: one each from 4T1 metastatic cell line variant #1, 4T1 variant #2, non-metastatic cell lines 4T07, and non-metastatic cell line 67NR. Classes were prepared in triplicate for a total of 12 samples. All samples were hybridized to the custom designed CFG GLYCOv2 glycogene array.

2011-02-17 | E-GEOD-27354 | biostudies-arrayexpress

Expression data of adult quiescent and activated mouse neural stem cells

Project description:Neural stem cells were sorted according to their activated or quiescent state by flow cytometry using a set of 3 markers (LeX, CD24 and EGFR) We used microarrays to detail the global programme of gene expression underlying the proliferation/quiescence balance.

2018-01-01 | GSE99777 | GEO

Transcriptomic profiles of D29, StarStuff and L5

Project description:The gene expression profiles of mycobacteriophage D29 and StarStuff were analyzed 15min, 30min, 60min and 150min post-infection (MOI=3), as well as a StarStuff lysogen. The gene expression profiles of 30min and 150min post-induction of an L5 thermo-inducible lysogen, as well as the L5 lysogen itself.

2017-11-13 | GSE99182 | GEO

Gene expression of Bacteroides thetaiotaomicron, in vivo with and without IgA 225.4

Project description:Rag1-/- C57BL/6 or Rag1-/- C57BL/6 injected with the IgA producing hybridoma producing 225.4 anti- Bacteroides thetaiotaomicron (Capsular polysaccharide 4 dependent carbohydrate epitope specific epitope) mice were colonized with B. thetaiotaomicron wildtype strain VPI-5482 for 10 days. Cecal bacteria were harvested and snap frozen and RNA isolated. Keywords: Single time point, experimental and control groups.

2007-12-27 | GSE9019 | GEO

Prdm16 newborn mouse (ventricular zone) VZ cells microarray

Project description:As Prdm16 deficiency reduces self-renewal potential and depletes neural stem cells in culture we decided to investigate the underlying molecular mechanisms of the neural stem cells depletion in the Prdm16 deficient animals. For the experiment we used Prdm16Gt(OST67423)Lex (Prdm16LacZ) genetrap mice obtained from the NIH Mutant Mouse Regional Resource Center (http://www.mmrrc.org/). We compared the gene expression profiles of uncultured ventricular zone cells from newborn Prdm16LacZ/LacZ (KO), Prdm16LacZ/+(HET), and Prdm16+/+ (WT) mice.

2010-09-09 | GSE23406 | GEO

CBP and p300 Jointly Maintain Neural Progenitor Viability but Play Unique Roles in the Differentiation of Neural Lineages

Project description:The paralogous lysine acetyltransferases 3 (KAT3), CBP and P300, play critical roles during neurodevelopment, but their specific roles in neural precursors maintenance and differentiation remain obscure. In fact, it is still unclear whether these proteins are individually or jointly essential in processes such as proliferation of neural precursors, differentiation to specific neural cell types, or both. Here, we use subventricular zone-derived neurospheres as a potential ex vivo developmental model to analyze the proliferation and differentiation of neural stem cells (NSCs) lacking CBP, p300, or both proteins. The results showed that CBP and p300 are not individually essential for maintenance and proliferation of NSCs, although their combined ablation seriously compromised cell division. In turn, the absence of either of the two proteins compromised the differentiation of NSC into the neuronal and astrocytic lineages. Single-nucleus RNA sequencing analysis of neural cell cultures derived from CBP or p300 mutant neurospheres revealed divergent trajectories of neural differentiation upon CBP or p300 ablation, confirming unique functions and nonredundant roles in neural development. These findings contribute to a better understanding of the shared and individual roles of KAT3 proteins in neural differentiation and the etiology of neurodevelopmental disorders caused by their deficiency.

2022-12-17 | GSE211737 | GEO

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