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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.

ORGANISM(S): Mus musculus Homo sapiens

PROVIDER: GSE27596 | GEO | 2011/03/02

SECONDARY ACCESSION(S): PRJNA138287

REPOSITORIES: GEO

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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. 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.

2011-03-02 | E-GEOD-27596 | biostudies-arrayexpress

To compare the levels of expression of the different glycosyltransferases in order to explain the exclusive detection of LeX in the neurons

Project description:Human NT2- cells differentiate in vitro into NT2N neurons in the presence of retinoic acid, and culture in the presence of mitosis inhibitors. We have previously observed the presence of the Lewisx determinant exclusively in NT2N neurons but not in NT2- cells. We also obtained evidence that Lex is synthesized in these cells by FUT9 (Brito,...,Costa (2007) J. Neurosci. Res. 85, 1260). We would like to compare the levels of expression of the different glycosyltransferases, particularly, fucosyltransferases and sialyltransferases to explain the exclusive detection of Lex in the neurons. In addition, we would like to know if there are some Lex-binding proteins that may act as binding partners of Lex in the neurons.

2011-10-02 | E-GEOD-32554 | 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

To compare the levels of expression of the different glycosyltransferases in order to explain the exclusive detection of LeX in the neurons

2011-10-03 | GSE32554 | GEO

Glycosyltransferases from oat (Avena) implicated in the acylation of avenacins

Project description:To study glycosyltransferases from oat, root proteins were extracted and separated gy 1D SDS gel electrophoresis. Bands were cut out, digested with trypsin, and the resulting peptides were analysed by LCMSMS on an Orbitrap mass sepctrometer. Raw data was processed with MaxQuant 1.3.0.5, and database searches on a custom database performed using Mascot.

2013-04-24 | PXD000083 | Pride

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

Mouse lymphocyte study

Project description:Dr. Paulson's group investigates the roles of carbohydrate-binding proteins involved in immune regulation and human disease, through their interaction with carbohydrate groups expressed on cell surface glycoconjugates. This laboratory is currently focused on how sialoside ligands modulate Siglec functions and on understanding the molecular basis of glycosylation changes following differentiation and activation of leukocytes. Evaluation of Glycosyltransferases and CBP expression in B cells, CD4+ and CD8+ T cells following activation. The overall goal of this study is to understand the molecular regulation of cell surface glycosylation in mouse lymphocytes before and after activation. Initial studies using lectins and antibodies showed differences in glycan expression following activation, which included a marked increase in PNA binding. Interestingly, this increase was observed in B, CD4+ and CD8+ cells after 72h of activation. Our aim is to elucidate the molecular basis for such changes in B and T lymphocytes (CD4+ and CD8+), focusing on the mRNA expression of glycogenes before and after activation of mixed spleenocytes. In brief, the experimental protocol is as follows: splenocytes are isolated from C57Bl/6 mice (6-10 wks old) and cultured for 0, 24, 48 and 72 hrs. Cultures containe either anti-CD3/IL2/IL4 or anti-IgM/IL4 to activate T cells and B cells respectively. At each time point, B, CD4+ and CD8+ cells are purified by positive selection using magnetic beads and total RNA is prepared from each cell type.

2011-01-07 | E-GEOD-26478 | biostudies-arrayexpress

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

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