Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy

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    Abstract

    To explore chorionic gonadotrophin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated in a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL after specific intervals of exposure (1, 3, 6 and 9 days) to recombinant hCG in vivo and hybridized to Affymetrix™ GeneChip Rhesus Macaque Genome Arrays. The mRNA levels of 1192 transcripts changed ≥2-fold [one-way ANOVA, false discovery rate (FDR) correction; P<0.05] during SEP when compared with Day 10 untreated controls. Real-time PCR validation indicated that 15 of 17 genes matched in expression pattern between PCR and microarray. Protein levels of three genes identified as CG-sensitive, CYP19A1 (aromatase), PGRMC1 (progestin-binding protein) and STAR (steroidogenic acute regulatory protein) were quantified by western blot analysis. To further analyze global changes in gene expression induced by CG exposure, luteal gene expression was compared between SEP (rescued) and regressing CL, utilizing previously banked GeneChip data from the luteal phase of the menstrual cycle. Expression patterns and mRNA levels were analyzed between time-matched intervals. Transcripts for 7677 mRNAs differed in expression patterns ≥2-fold (one-way ANOVA, FDR correction; P<0.05) between the hCG-exposed (SEP) CL and regressing CL. Regressed CL (at menses) were most unlike all other CL. Pathway analysis of significantly affected transcripts was performed; the pathway most impacted by CG exposure was steroid biosynthesis. Further comparisons of the genome-wide changes in luteal gene expression during CG rescue and luteolysis in the natural menstrual cycle should identify additional key regulatory pathways promoting primate fertility.

    Original languageEnglish (US)
    Article numbergar073
    Pages (from-to)216-227
    Number of pages12
    JournalMolecular Human Reproduction
    Volume18
    Issue number4
    DOIs
    StatePublished - Apr 2012

    Fingerprint

    Corpus Luteum
    Chorionic Gonadotropin
    Microarray Analysis
    Transcriptome
    Primates
    Pregnancy
    Gene Expression
    Macaca mulatta
    Messenger RNA
    Analysis of Variance
    Genome
    Luteolysis
    Aromatase
    Menstruation
    Luteal Phase
    Progestins
    Menstrual Cycle
    Genes
    Fertility
    Real-Time Polymerase Chain Reaction

    Keywords

    • Chorionic gonadotrophin
    • Corpus luteum
    • Early pregnancy
    • Microarray

    ASJC Scopus subject areas

    • Molecular Biology
    • Embryology
    • Cell Biology
    • Genetics
    • Developmental Biology
    • Reproductive Medicine
    • Obstetrics and Gynecology

    Cite this

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    title = "Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy",
    abstract = "To explore chorionic gonadotrophin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated in a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL after specific intervals of exposure (1, 3, 6 and 9 days) to recombinant hCG in vivo and hybridized to Affymetrix™ GeneChip Rhesus Macaque Genome Arrays. The mRNA levels of 1192 transcripts changed ≥2-fold [one-way ANOVA, false discovery rate (FDR) correction; P<0.05] during SEP when compared with Day 10 untreated controls. Real-time PCR validation indicated that 15 of 17 genes matched in expression pattern between PCR and microarray. Protein levels of three genes identified as CG-sensitive, CYP19A1 (aromatase), PGRMC1 (progestin-binding protein) and STAR (steroidogenic acute regulatory protein) were quantified by western blot analysis. To further analyze global changes in gene expression induced by CG exposure, luteal gene expression was compared between SEP (rescued) and regressing CL, utilizing previously banked GeneChip data from the luteal phase of the menstrual cycle. Expression patterns and mRNA levels were analyzed between time-matched intervals. Transcripts for 7677 mRNAs differed in expression patterns ≥2-fold (one-way ANOVA, FDR correction; P<0.05) between the hCG-exposed (SEP) CL and regressing CL. Regressed CL (at menses) were most unlike all other CL. Pathway analysis of significantly affected transcripts was performed; the pathway most impacted by CG exposure was steroid biosynthesis. Further comparisons of the genome-wide changes in luteal gene expression during CG rescue and luteolysis in the natural menstrual cycle should identify additional key regulatory pathways promoting primate fertility.",
    keywords = "Chorionic gonadotrophin, Corpus luteum, Early pregnancy, Microarray",
    author = "Cecily Bishop and S. Satterwhite and L. Xu and Jon Hennebold and Richard Stouffer",
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    AU - Bishop, Cecily

    AU - Satterwhite, S.

    AU - Xu, L.

    AU - Hennebold, Jon

    AU - Stouffer, Richard

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    AB - To explore chorionic gonadotrophin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated in a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL after specific intervals of exposure (1, 3, 6 and 9 days) to recombinant hCG in vivo and hybridized to Affymetrix™ GeneChip Rhesus Macaque Genome Arrays. The mRNA levels of 1192 transcripts changed ≥2-fold [one-way ANOVA, false discovery rate (FDR) correction; P<0.05] during SEP when compared with Day 10 untreated controls. Real-time PCR validation indicated that 15 of 17 genes matched in expression pattern between PCR and microarray. Protein levels of three genes identified as CG-sensitive, CYP19A1 (aromatase), PGRMC1 (progestin-binding protein) and STAR (steroidogenic acute regulatory protein) were quantified by western blot analysis. To further analyze global changes in gene expression induced by CG exposure, luteal gene expression was compared between SEP (rescued) and regressing CL, utilizing previously banked GeneChip data from the luteal phase of the menstrual cycle. Expression patterns and mRNA levels were analyzed between time-matched intervals. Transcripts for 7677 mRNAs differed in expression patterns ≥2-fold (one-way ANOVA, FDR correction; P<0.05) between the hCG-exposed (SEP) CL and regressing CL. Regressed CL (at menses) were most unlike all other CL. Pathway analysis of significantly affected transcripts was performed; the pathway most impacted by CG exposure was steroid biosynthesis. Further comparisons of the genome-wide changes in luteal gene expression during CG rescue and luteolysis in the natural menstrual cycle should identify additional key regulatory pathways promoting primate fertility.

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