title: Research Resource: The Dexamethasone Transcriptome in Hypothalamic Embryonic Neural Stem Cells.
abstract: Exposure to excess glucocorticoids during fetal development has long-lasting physiological and behavioral consequences, although the mechanisms are poorly understood. The impact of prenatal glucocorticoids exposure on stress responses in juvenile and adult offspring implicates the developing hypothalamus as a target of adverse prenatal glucocorticoid action. Therefore, primary cultures of hypothalamic neural-progenitor/stem cells (NPSCs) derived from mouse embryos (embryonic day 14.5) were used to identify the glucocorticoid transcriptome in both males and females. NPSCs were treated with vehicle or the synthetic glucocorticoid dexamethasone (dex; 100nM) for 4 hours and total RNA analyzed using RNA-Sequencing. Bioinformatic analysis demonstrated that primary hypothalamic NPSC cultures expressed relatively high levels of a number of genes regulating stem cell proliferation and hypothalamic progenitor function. Interesting, although these cells express glucocorticoid receptors (GRs), only low levels of sex-steroid receptors are expressed, which suggested that sex-specific differentially regulated genes identified are mediated by genetic and not hormonal influences. We also identified known or novel GR-target coding and noncoding genes that are either regulated equivalently in male and female NPSCs or differential responsiveness in one sex. Using gene ontology analysis, the top functional network identified was cell proliferation and using bromodeoxyuridine (BrdU) incorporation observed a reduction in proliferation of hypothalamic NPSCs after dexamethasone treatment. Our studies provide the first characterization and description of glucocorticoid-regulated pathways in male and female embryonically derived hypothalamic NPSCs and identified GR-target genes during hypothalamic development. These findings may provide insight into potential mechanisms responsible for the long-term consequences of fetal glucocorticoid exposure in adulthood.
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Krystle A
authors.initials: KA
authors.lastname: Frahm
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Melanie E
authors.initials: ME
authors.lastname: Peffer
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Janie Y
authors.initials: JY
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Soumya
authors.initials: S
authors.lastname: Luthra
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Anish B
authors.initials: AB
authors.lastname: Chakka
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Couger
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Uma R
authors.initials: UR
authors.lastname: Chandran
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: A Paula
authors.initials: AP
authors.lastname: Monaghan
authors.affiliation: Department of Pharmacology and Chemical Biology (K.A.F., J.Y.Z., D.B.D.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; Program in Integrative Molecular Biology (M.E.P., D.B.D.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Biomedical Informatics (S.L., A.B.C., U.R.C.), University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Microbiology and Molecular Genetics (M.B.C.), Oklahoma State University, Stillwater, Oklahoma; and Department of Neurobiology (A.P.M.), University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Donald B
authors.initials: DB
authors.lastname: DeFranco
mesh_terms: D000818:Animals; D003907:Dexamethasone / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005938:Glucocorticoids / Q000494:pharmacology*; D007031:Hypothalamus / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D005938:Glucocorticoids; D003907:Dexamethasone