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Identify molecules regulated by glucosylceramide synthase in mouse osteoblasts.

ABSTRACT: To identify molecules regulated by glucosylceramide synthase in osteoblasts, we conducted microarray analysis using samles treated with the inhibitors of glucosylceramide synthase.

ORGANISM(S): Mus musculus

PROVIDER: GSE174323 | GEO | 2021/08/30

REPOSITORIES: GEO

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Identify molecules regulated by glucosylceramide synthase in mouse osteoblasts.

Project description:Identify molecules regulated by glucosylceramide synthase in mouse osteoblasts.

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Contribution of Glucosylceramide Synthase to the Proliferation of Mouse Osteoblasts.

Project description:Background/aimGlycosphingolipids are known to be involved in bone metabolism. However, their roles and regulatory mechanisms in osteoblast proliferation are largely unknown. In this study, we examined the effects of inhibitors of glucosylceramide synthase (GCS), which is responsible for the generation of all glycosphingolipids, on osteoblast proliferation.Materials and methodsWe analyzed the expression of glycosphingolipids and cell growth in MC3T3-E1 mouse osteoblast cells treated with the GCS inhibitors miglustat, D-PDMP and D-PPMP. We also conducted microarray analysis and RNA interference to identify genes involved in cell growth regulated by GCS.ResultsGlycosphingolipids GD1a and Gb4 expressed in MC3T3-E1 cells, were suppressed by GCS inhibitors. Furthermore, the proliferation of MC3T3-E1 cells was suppressed by the inhibitors. Using microarray analysis, we predicted nine genes (Fndc1, Acta2, Igfbp5, Cox6a2, Cth, Mymk, Angptl6, Mab21l2, and Igsf10) suppressed by all three inhibitors. Furthermore, partial silencing of Angptl6 by RNA interference reduced MC3T3-E1 cell growth.ConclusionThese results show that GCS regulates proliferation through Angptl6 in osteoblasts.

| S-EPMC8627765 | biostudies-literature

Glucosylceramide fed mice skin before SDS treatment (0d), after SDS treatment (2d)

Project description:Dietary glucosylceramide (GC) improves skin barrier function. To elucidate the molecular mechanisms involved, we used a microarray system to analyze mRNA expression in SDS-treated dorsal skin of hairless mouse. Transepidermal water loss of mouse skin was increased by SDS treatment and the increase was significantly reduced by prior oral administration of glucosylceramides. Microarray-evaluated mRNA expression ratios showed statistically significant increase of expression of genes related to cornified envelope and tight junction formation versus all genes in glucosylceramide-fed/SDS-treated mouse skin. We then examined the contribution of glucosylceramide metabolites to tight junction formation of cultured keratinocytes. SDS treatment of cultured keratinocytes significantly decreased the transepidermal electrical resistance, and the decrease was significantly ameliorated in the presence of sphingosine or phytosphingosine, the major metabolites of glucosylceramide. These results suggest that oral administration of glucosylceramide improves skin barrier function by upregulating genes associated with both cornified envelope and tight junction formation. Two-condition experiment, effect of oral intake of GC on mice skin before SDS treatment (0d), and after SDS treatment(2d), Biological replicates: 3 replicates for 0d, 4 replicates for 2d.

2012-03-31 | E-GEOD-28086 | biostudies-arrayexpress

Transcriptome sequencing data sets for mice brain infected with Sindbis virus and treated with glucosylceramide synthase inhibitor

Project description:Transcriptome sequencing data sets for mice brain infected with Sindbis virus and treated with glucosylceramide synthase inhibitor

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Glucosylceramide fed mice skin before SDS treatment (0d), after SDS treatment (2d)

2012-04-01 | GSE28086 | GEO

Transcriptome sequencing data sets for mice brain infected with Sindbis virus and treated with glucosylceramide synthase inhibitor

Project description:The goal of this study is to determine the brain pathology upon infection of mice with neuroinvasine Sindbis virus (SVNI) and the affect of treatmant wuth the glucosylceramide inhibitor GZ-161 . For this purpose, C57BL/6 mice wasinfected with SVNI and treted or not with GZ-161. 5 days post infection, brains were harvested and total RNA was extracted. RNA-seq libraries were constructed and sequencing of 100bp paired-end was performed on the Illumina NovaSeq 6000 system. Sequencing yielded about 30M reads per sample that were mapped to the mouse genome

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Effect of Shiga toxin (Stx) and glucosylceramide synthase inhibitor (Gb3i) treatments on SV40 transformed human kidney endothelial cells

Project description:Effect of Shiga toxin (Stx) and glucosylceramide synthase inhibitor (Gb3i) treatments on SV40 transformed human kidney endothelial cells

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Complex formation of sphingomyelin synthase 1 with glucosylceramide synthase increases sphingomyelin and decreases glucosylceramide levels.

Project description:Sphingolipids, including sphingomyelin (SM) and glucosylceramide (GlcCer), are generated by the addition of a polar head group to ceramide (Cer). Sphingomyelin synthase 1 (SMS1) and glucosylceramide synthase (GCS) are key enzymes that catalyze the conversion of Cer to SM and GlcCer, respectively. GlcCer synthesis has been postulated to occur mainly in cis-Golgi, and SM synthesis is thought to occur in medial/trans-Golgi; however, SMS1 and GCS are known to partially co-localize in cisternae, especially in medial/trans-Golgi. Here, we report that SMS1 and GCS can form a heteromeric complex, in which the N terminus of SMS1 and the C terminus of GCS are in close proximity. Deletion of the N-terminal sterile α-motif of SMS1 reduced the stability of the SMS1-GCS complex, resulting in a significant reduction in SM synthesis in vivo In contrast, chemical-induced heterodimerization augmented SMS1 activity, depending on an increase in the amount and stability of the complex. Fusion of the SMS1 N terminus to the GCS C terminus via linkers of different lengths increased SM synthesis and decreased GlcCer synthesis in vivo These results suggest that formation of the SMS1-GCS heteromeric complex increases SM synthesis and decreases GlcCer synthesis. Importantly, this regulation of relative Cer levels by the SMS1-GCS complex was confirmed by CRISPR/Cas9-mediated knockout of SMS1 or GCS combined with pharmacological inhibition of Cer transport protein in HEK293T cells. Our findings suggest that complex formation between SMS1 and GCS is part of a critical mechanism controlling the metabolic fate of Cer in the Golgi.

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Screen mRNAs specifically regulated by mTORC1 in calvarial osteoblasts from mice

Project description:To screen mRNAs specifically regulated by mTORC1, a global mRNA expression profile in calvarial osteoblasts (OBs) from mice with or without OB-specific Tsc1 knockout was developed using microarray. Wild type (WT) or OB-specific Tsc1 knockout (KO) mice were sacrificed, with calvarial osteoblasts harvested and subjected to total RNA extraction.

2015-11-07 | E-GEOD-74781 | biostudies-arrayexpress

Identification of miRNAs regulated by vitamin D within primary human osteoblasts

Project description:Identified a series of miRNAs transcriptionally regulated by vitamin D treatment within osteoblasts. Several miRNAs were validated in dose escalation experiments. Furthermore, downstream miRNA targets were appraised and found to regulate target message and protein levels. These findings were assessed in early and late passaged osteoblasts and found an age-dependent regulation of some miRNAs, while others were considered constituitiously expressed. Inhibitor studies of one key bone-related miRNA demonstrated the importance of this miRNA to regulate vitamin D-mediate osteoblastic differentiation during mineralizaition. Three independent sets of passage 8 primary human osteoblasts were either treated with vehicle (ethanol) or 1,25D3 (10nM) for 6 hours. Total RNA was collected using the miRNA Vana kit (ABI) and then quality assessed, and sent for miRNA array processesing.

2012-07-31 | E-GEOD-34144 | biostudies-arrayexpress

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