Single Cell RNA Sequencing of fetal mouse neurosphere cultures.
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE155880 | GEO | 2021/02/10
REPOSITORIES: GEO
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Project description:Pregannt Cyp19-EGFP mice (maintained on a soybean free mouse chow diet) were treated with either 50μg/kg/day Bsiphenol A or vehicle (ethinol in peanut oil) between gestational days (GD) 10.5-14.5. On GD15.5, fetuses were dissected and sex determined by SRY genotyping. Fetuses were obtained from different dams.
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Project description:Single Cell RNA Sequencing of fetal mouse neurosphere cultures.
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Project description:The expression of the mciroRNA, miR-153 is significantly altered in neural stem cells (NSCs) following exposure to the teratogens, alcohol and nicotine (PMIDs:22458409, 17687032). To better understand how miR-153 may mediate teratogenesis, we first needed to identify miR153 targets in fetal NSCs. Gestational day 12.5 fetal mouse cortical neural epithelium was isolated and grown as neurospheres in defined medium for studies. We overexpressed miR-153 in fetal NSCs for 24 hours in a transient transfection assay, and profiled mRNA expression using a microarray platform (codelink array) to identify potential target genes. Over expression of pre-miR-153 resulted in a ~32-fold induction of miR-153 compared to the transfection control. Transcript profiles were assessed from RNA samples (6 independent replicates in each condition) hybridized to codelink microarrays according to manufacturer's protocols. Data analysis showed that miR-153 overexpression resulted in statistically significant down-regulation of 102 genes (at a false discovery rate α=0.1, of a total of 860 transcripts down-regulated by ≥1.3-fold). These data suggested the role of this miRNA in moderately controlling expression hundreds of gene transcripts. These regulated genes may be important in controlling neural stem cell differentiation and downstream signaling pathways.
2014-08-01 | GSE49684 | GEO
Project description:The expression of the mciroRNA, miR-153 is significantly altered in neural stem cells (NSCs) following exposure to the teratogens, alcohol and nicotine (PMIDs:22458409, 17687032). To better understand how miR-153 may mediate teratogenesis, we first needed to identify miR153 targets in fetal NSCs. Gestational day 12.5 fetal mouse cortical neural epithelium was isolated and grown as neurospheres in defined medium for studies. We overexpressed miR-153 in fetal NSCs for 24 hours in a transient transfection assay, and profiled mRNA expression using a microarray platform (codelink array) to identify potential target genes. Over expression of pre-miR-153 resulted in a ~32-fold induction of miR-153 compared to the transfection control. Transcript profiles were assessed from RNA samples (6 independent replicates in each condition) hybridized to codelink microarrays according to manufacturer's protocols. Data analysis showed that miR-153 overexpression resulted in statistically significant down-regulation of 102 genes (at a false discovery rate α=0.1, of a total of 860 transcripts down-regulated by ≥1.3-fold). These data suggested the role of this miRNA in moderately controlling expression hundreds of gene transcripts. These regulated genes may be important in controlling neural stem cell differentiation and downstream signaling pathways. Six independent replicates were performed for control and treatment groups, and twelve independent single channel intensity values were uploaded after normalization for analysis independent single channel intensity values were uploaded after normalization
2014-08-01 | E-GEOD-49684 | biostudies-arrayexpress
Project description:To understand the mechanism by which miR-335 acts anti proliferative factor, we profiled mRNA expression using microarray platform ( codelink array). Embryonic day 12.5 neural cortical epithelium was isolated and grown as neurospheres in defined medium. Cultures were transfected with 2'-O-methyl (2'OMe)-base modified miRNA-335 antisense (2'OMe-335) and control (2' O Me-EGFP). To verify the suppression of miR-335 after the 2'-OMe treatment, we used real time RT-PCR which confirmed the down regulation of miR-335 up to ~4.5 cycles (~24 fold). For the microarray experiment, mRNA isolated from these cells was hybridized to codelink microarrays. Three independent replicate were done for treatment and control, six independent single channel intensity values were uploaded after normalization for analysis. Our array results indicated that there is general up-regulation in gene expression due to miR-335 Knock Out ( KO). We identified 116 genes statistically significant genes (adjusted p value < 0.05), out of these 95.7% were up-regulated genes. The highest fold difference we observed was between 2 and 3 fold, indicating the role of this miRNA in moderately controlling expression hundreds of genes. Of the several genes the highly expressed are important in maturation of neural stem cells and downstream signaling pathway genes
2019-07-31 | GSE14624 | GEO
Project description:Knowing the gene expression profiles of drug-metabolizing enzymes and transporters throughout gestation is important for understanding the mechanisms of pregnancy-induced changes in drug pharmacokinetics. In this study, we compared gene expression of drug-metabolizing enzymes and transporters in the maternal liver, kidney, small intestine, and placenta of pregnant mice throughout gestation by microarray analysis. Specifically, we investigated cytochrome P450 (Cyp), UDP-glucuronosyltranserase (Ugt), and sulfotransferase (Sult), as well as ATP-binding cassette (Abc) and solute carrier (Slc) transporters. We found that relatively few Ugt and Sult genes were impacted by pregnancy in maternal tissues and placenta. Cyp1a2, most Cyp2 isoforms, Cyp3a11, and Cyp3a13 in the liver were down-regulated, with the greatest changes occurring on gestation days (gd) 15 and 19 compared to non-pregnant controls (gd 0). However, Cyp2d40, Cyp3a16, Cyp3a41a, Cyp3a41b, and Cyp3a44 in the liver were induced throughout pregnancy. Cyp expression in mid-gestation placenta (gd 10 and 15) was generally greater than that in term placenta (gd 19). There were also notable changes in Abc and Slc transporters. Abcc3 in the liver was down-regulated by 60%, and Abcb1a, Abcc4, and Slco4c1 in the kidney were down-regulated by 30-60% on gd 15 and 19 versus gd 0. Abcc5 in the placenta was induced 3-fold on gd 10 versus gd 15 and 19, whereas Slc22a3 expression in the placenta on gd 10 was 90% lower than that on gd 15 and 19. Overall, this study demonstrates important gestational age-dependent expression of drug-metabolizing enzymes and transporter genes, which may have mechanistic relevance to human pregnancy. Ninety pregnant mice at gestational days 0, 7.5, 10, 15, and 19 (n = 5-6 per gestational age) were used for the maternal liver, kidney, small intestine and placenta. The placentas were collected on gestational days 10, 15, and 19.
2013-10-08 | E-GEOD-41438 | biostudies-arrayexpress
Project description:The Long Evans/orl (LE/orl) rat is an animal model of inherited undescended testis (UDT). To explore genetic mechanisms of UDT, we studied differential gene expression in LE/orl and LE wild type (LE/wt) fetal gubernaculum and testis. Analyses of differentially expressed probe sets linked below as supplementary files:; Analysis_methods.txt [A description of the analysis]; gub.txt [Genes differentially expressed in between wt and orl gubernacula]; testes.txt [Genes differentially expressed between wt and orl testes]; wt.txt [Genes differentially expressed in wt gubernacular development] Experiment Overall Design: Fetuses were harvested from timed matings of LE/wt and LE/orl rats at gestational day (GD) 17, 18, 19, and 20. Single gubernaculum and testis were removed from male fetuses and total RNA was isolated from each organ. Left organs were used, except for GD 20 fetuses where both right and left gubernacula were included. For female fetuses, the right and left gubernacula from a single fetus were pooled before RNA isolation. A total of 54 samples were hybridized to GeneChips; 34 gubernacular samples and 20 testes samples. There were 13 sample groups, each included biological replicate arrays. The sample groups were (1) female LE/wt GD18 gubernacula n=4, (2) male LE/wt GD18 left gubernacula n=4, (3) male LE/wt GD19 left gubernacula n=4, (4) male LE/wt GD20 left gubernacula n=4, (5) male LE/wt GD20 right gubernacula n=3, (6) male LE/orl GD18 left gubernacula n=4, (7) male LE/orl GD19 left gubernacula n=4, (8) male LE/orl GD20 left gubernacula n=4, (9) male LE/orl GD20 right gubernacula n=3, (10) male LE/wt GD17 left testes n=5, (11) male LE/wt GD19 left testes n=5, (12) male LE/orl GD17 left testes n=5, and (13) male LE/orl GD19 left testes n=5.
2008-06-15 | E-GEOD-7755 | biostudies-arrayexpress
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