Progesterone Regulation of Extracellular Vesicles in the Uterine Luminal Fluid of Sheep (RNA-Seq)
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ABSTRACT:
ORGANISM(S): Ovis aries
PROVIDER: GSE103593 | GEO | 2017/09/08
SECONDARY ACCESSION(S): PRJNA402035
REPOSITORIES: GEO
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Project description:Progesterone (P4) acts via the endometrium to modify the uterine environment and promotes conceptus growth for elongation and pregnancy establishment. Ewes were ovariectomized and treated with P4 for 14 days or P4 for 14 days and RU486, a progesterone receptor antagonist, from days 8 to 14. Small RNA sequencing of endometrium and EVs from the uterine lumen detected expression of 768 miRNAs and P4 regulation of 9 endometrial and 7 extracellular vesicle miRNAs.
2017-09-08 | GSE103595 | GEO
Project description:Here we used RNAseq to assess the impact of progesterone (P4) on the transcriptome of the medio-basal hypothalamus of ewes. Ewes were treated with a vaginal fluogestone acetate sponge in mid-February. The sponge was removed in early March and ewes received a PMSG injection. Animals were then killed either 2 days (follicular phase) or 12 days (luteal phase) later.
2022-05-12 | GSE202508 | GEO
Project description:Ovine interferon-tau (IFNT) is released from the conceptus by Day 12 of pregnancy and disrupts pulsatile release of endometrial prostaglandin F2 alpha (PGF), thereby protecting the corpus luteum (CL). IFNT may also have endocrine action through inducing interferon stimulated genes (ISGs) in the CL. The hypothesis that gene expression differs in CL collected from pregnant (P) and non-pregnant (NP) ewes by Day 14 due to the lytic action of PGF during the estrous cycle or the presence of a conceptus was tested. RNA was isolated on Days 12 and 14 in NP or P ewes (n = 3 ewes/group) and analyzed using the Affymetrix bovine microarray (24,000 targets). Differential gene expression (>1.5 fold, P < 0.05) was confirmed using semi-quantitative real time PCR (RTPCR). Serum progesterone concentrations decreased (P < 0.05) from 1.7 ng/ml on Day 12 to 1.3 ng/ml by Day 14 in NP ewes suggesting initiation of luteolysis; and remained > 1.7 ng/ml in Day 12 and 14 P ewes indicating that the conceptus protected the CL from luteolysis. Early luteolysis from Day 12 to 14 NP was associated with differential expression of 683 genes, including SERPINE1 and THBS1. Presence of a conceptus from Day 12 to 14 also induced expression of 743 genes, i.e., ISGs (ISG15, MX1), PTX3, and IL-6 and stabilized expression of VEGF and LHR genes. In conclusion, pregnancy circumvents luteolytic pathways, and activates or stabilizes genes associated with interferon, chemokine, cell adhesion, cytoskeletal, angiogenic and epithelial to mesynchymal transition pathways in the CL. There are 12 samples that were analyzed for the microarray, no duplicates and we increased the sample size to 50 for the RTPCR. This Series contains the Affymetrix array data only (not RT-PCR data).
2013-11-01 | E-GEOD-47776 | biostudies-arrayexpress
Project description:Ovine interferon-tau (IFNT) is released from the conceptus by Day 12 of pregnancy and disrupts pulsatile release of endometrial prostaglandin F2 alpha (PGF), thereby protecting the corpus luteum (CL). IFNT may also have endocrine action through inducing interferon stimulated genes (ISGs) in the CL. The hypothesis that gene expression differs in CL collected from pregnant (P) and non-pregnant (NP) ewes by Day 14 due to the lytic action of PGF during the estrous cycle or the presence of a conceptus was tested. RNA was isolated on Days 12 and 14 in NP or P ewes (n = 3 ewes/group) and analyzed using the Affymetrix bovine microarray (24,000 targets). Differential gene expression (>1.5 fold, P < 0.05) was confirmed using semi-quantitative real time PCR (RTPCR). Serum progesterone concentrations decreased (P < 0.05) from 1.7 ng/ml on Day 12 to 1.3 ng/ml by Day 14 in NP ewes suggesting initiation of luteolysis; and remained > 1.7 ng/ml in Day 12 and 14 P ewes indicating that the conceptus protected the CL from luteolysis. Early luteolysis from Day 12 to 14 NP was associated with differential expression of 683 genes, including SERPINE1 and THBS1. Presence of a conceptus from Day 12 to 14 also induced expression of 743 genes, i.e., ISGs (ISG15, MX1), PTX3, and IL-6 and stabilized expression of VEGF and LHR genes. In conclusion, pregnancy circumvents luteolytic pathways, and activates or stabilizes genes associated with interferon, chemokine, cell adhesion, cytoskeletal, angiogenic and epithelial to mesynchymal transition pathways in the CL.
2013-11-01 | GSE47776 | GEO
Project description:The goal of this study is to analyze changes in gene expression produced in posterior midguts after knocking down Gli in enterocytes (EC) during 9 days. Gli was depleted from ECs using a drug-inducible version of the binary GAL4-UAS system in combination with UAS-GLI-RNAi. Targeted gene expression using the 5966GS-GAL4 driver is observed in adult ECs when flies are fed the progesterone analog RU486 (RU+); addition of ethanol is used as a control (RU-), providing cohorts of isogenic individuals with or without induction of the Gli-RNAi transgene.
2016-12-18 | GSE74168 | GEO
Project description:ovariectomized mice treated with E2+Progesterone, E2+Progesterone+TPA, E2+Progesterone+RU486 and sham for 14 days, followed by isolation of luminal mature, luminal progenitor and mammary stem cells through FACS and RNASeq performed for each cell lineage population. TPA and RU486 treated mice show significant decline in mammary stem cell pool pulation comapared to EP treated mice
2020-02-26 | GSE127197 | GEO
Project description:The myometrium is an important reproductive tissue composed primarily of smooth muscle cells. Contractility of the myometrial smooth muscle cells during pregnancy and labour is modulated by hormones. Despite much research, little is known about the molecular mechanism by which estrogen and progesterone regulate myometrial contractility. This study investigates global gene expression profile of cultured human uterine smooth muscle cells (hUtSMCs) following 17β-estradiol (E2) and/or progesterone treatments using cDNA microarray technology. Furthermore the effects of addition of the progesterone inhibitor RU486 were investigated. Many genes were regulated in the presence of P4 or E2 alone but almost six times these numbers were regulated in the presence of the combination. The majority of these genes returned to near baseline levels (control samples) on addition of RU486. In total 796 annotated genes were significantly differentially expressed in the combined presence of P4 and E2 (relative to untreated levels). Furthermore, 666 annotated genes were significantly regulated by the addition of RU486 to the P4/E2 combination (relative to P4/E2 levels). Gene ontology analysis of these differentially expressed genes revealed a strong association of P4/E2 treatment with the downregulated expression of genes involved in cell communication, signal transduction, channel activity, inflammatory response and differentiation. Upregulated processes included cell survival (anti-apoptosis), gene transcription, steroid hormone biosynthesis, muscle development, insulin receptor signaling and cell growth. Functional withdrawal of progesterone using RU486 effectively reversed the processes induced by P4/E2 treatment. The hUtSMC system is an additional useful model to investigate steroid effects on smooth muscle cells in isolation from other myometrial cell types.
2015-11-02 | GSE59231 | GEO
Project description:The myometrium is an important reproductive tissue composed primarily of smooth muscle cells. Contractility of the myometrial smooth muscle cells during pregnancy and labour is modulated by hormones. Despite much research, little is known about the molecular mechanism by which estrogen and progesterone regulate myometrial contractility. This study investigates global gene expression profile of cultured human uterine smooth muscle cells (hUtSMCs) following 17β-estradiol (E2) and/or progesterone treatments using cDNA microarray technology. Furthermore the effects of addition of the progesterone inhibitor RU486 were investigated. Many genes were regulated in the presence of P4 or E2 alone but almost six times these numbers were regulated in the presence of the combination. The majority of these genes returned to near baseline levels (control samples) on addition of RU486. In total 796 annotated genes were significantly differentially expressed in the combined presence of P4 and E2 (relative to untreated levels). Furthermore, 666 annotated genes were significantly regulated by the addition of RU486 to the P4/E2 combination (relative to P4/E2 levels). Gene ontology analysis of these differentially expressed genes revealed a strong association of P4/E2 treatment with the downregulated expression of genes involved in cell communication, signal transduction, channel activity, inflammatory response and differentiation. Upregulated processes included cell survival (anti-apoptosis), gene transcription, steroid hormone biosynthesis, muscle development, insulin receptor signaling and cell growth. Functional withdrawal of progesterone using RU486 effectively reversed the processes induced by P4/E2 treatment. The hUtSMC system is an additional useful model to investigate steroid effects on smooth muscle cells in isolation from other myometrial cell types. In this study human myometrial smooth muscle cells (hUtSMCs) purchased from Lonza were cultured and treated with 17beta estradiol and progesterone either alone or in combination. In addition the 17beta estradiol and progesterone combined treated samples were also treated with the progesterone inhibitor RU486 (mifepristone). RNA was isolated after 72h from both treated and untreated control cells. The experiment was repeated 3 times. Please note that slides were scanned three times successively (i.e. 3 .gpr files per sample) and the median raw values were used as input raw data for GeneSpring analysis (Median_Table_InputRAW.xlsx).
2015-11-02 | E-GEOD-59231 | biostudies-arrayexpress
Project description:An increase in circulating progesterone (P4) concentrations is associated with increased pregnancy success in beef and dairy cattle. Our objective was to ascertain differential effects of elevated P4 concentrations following conception on endometrial gene expression in beef heifers on Days 5, 7, 13 and 16 of pregnancy, corresponding to the morula, blastocyst, elongation and maternal recognition of pregnancy stages, respectively. Estrus was synchronized in beef heifers (N=263). Two-thirds (N=140) were inseminated (Day 0), and all animals were randomly assigned to one of the following treatments: (i) pregnant, high P4; (ii) pregnant, normal P4; (iii) cycling, high P4; (iv) and cycling, normal P4. All high P4 groups received a P4 release intravaginal device (PRID) on Day 3 post-estrus/mating. Tissue was collected on Days 5, 7, 13 or 16 of the cycle or pregnancy, and pregnancy was confirmed by the presence of an appropriately developed embryo/conceptus. PRID insertion elevated (P<0.05) P4 concentrations from Day 3.5 to 8 compared with untreated animals and conceptus size was larger (P<0.05) in animals with elevated P4 on Days 13 and 16 compared with normal P4. Total RNA was extracted from predominantly intercaruncular endometria from the ipsilateral uterine horn. Samples from individual heifers were selected on the basis of their P4 profiles and gene expression was analyzed using bovine Affymetrix microarrays (N=5 per treatment per time point). Microarray data from analyses using Bioconductor GCRMA and Limma packages were subjected to a modified t-test and P-values were adjusted for multiple testing using the Benjamin and Hochberg false discovery rate method. Differentially expressed genes were selected on the basis of an adjusted P-value of <0.01. There were no detectable differences in gene expression in endometria from pregnant and cyclic heifers on Days 5, 7 and 13 post-estrus, but, the expression of 764 genes was altered due to the presence of the conceptus at maternal recognition of pregnancy (Day 16). On Days 5 and 7, elevated P4 in pregnant heifers, altered the expression of 36 and 124 genes respectively but on Days 13 and 16 there were relatively few DEG between high and normal P4 heifers (15 and 25). Of the genes that were differentially regulated by P4, the majority were unique to a specific day of the estrous cycle/early pregnancy. In conclusion, gene expression in endometria did not differ between pregnant and cycling heifers until Day 16 of pregnancy (i.e. the time of maternal recognition of pregnancy and production of interferon tau by conceptus trophectoderm); however, elevating P4 in early pregnancy programmed changes in gene expression in endometria that are hypothesized to impact early conceptus growth and development. Thus, on Days 5, 7 and 13 differential gene expression was affected by P4, but on Day 16 the conceptus primarily influenced gene expression in uterine endometria of heifers. Endometiral samples were taken from estrous synchronized beef heifers that were either cyclic or confirmed pregnant with either normal or high concentrations of progesterone on Day 5, 7, 13 and 16 of the estrous cycle/early pregnancy.
2011-10-17 | E-GEOD-33030 | biostudies-arrayexpress
Project description:An increase in circulating progesterone (P4) concentrations is associated with increased pregnancy success in beef and dairy cattle. Our objective was to ascertain differential effects of elevated P4 concentrations following conception on endometrial gene expression in beef heifers on Days 5, 7, 13 and 16 of pregnancy, corresponding to the morula, blastocyst, elongation and maternal recognition of pregnancy stages, respectively. Estrus was synchronized in beef heifers (N=263). Two-thirds (N=140) were inseminated (Day 0), and all animals were randomly assigned to one of the following treatments: (i) pregnant, high P4; (ii) pregnant, normal P4; (iii) cycling, high P4; (iv) and cycling, normal P4. All high P4 groups received a P4 release intravaginal device (PRID) on Day 3 post-estrus/mating. Tissue was collected on Days 5, 7, 13 or 16 of the cycle or pregnancy, and pregnancy was confirmed by the presence of an appropriately developed embryo/conceptus. PRID insertion elevated (P<0.05) P4 concentrations from Day 3.5 to 8 compared with untreated animals and conceptus size was larger (P<0.05) in animals with elevated P4 on Days 13 and 16 compared with normal P4. Total RNA was extracted from predominantly intercaruncular endometria from the ipsilateral uterine horn. Samples from individual heifers were selected on the basis of their P4 profiles and gene expression was analyzed using bovine Affymetrix microarrays (N=5 per treatment per time point). Microarray data from analyses using Bioconductor GCRMA and Limma packages were subjected to a modified t-test and P-values were adjusted for multiple testing using the Benjamin and Hochberg false discovery rate method. Differentially expressed genes were selected on the basis of an adjusted P-value of <0.01. There were no detectable differences in gene expression in endometria from pregnant and cyclic heifers on Days 5, 7 and 13 post-estrus, but, the expression of 764 genes was altered due to the presence of the conceptus at maternal recognition of pregnancy (Day 16). On Days 5 and 7, elevated P4 in pregnant heifers, altered the expression of 36 and 124 genes respectively but on Days 13 and 16 there were relatively few DEG between high and normal P4 heifers (15 and 25). Of the genes that were differentially regulated by P4, the majority were unique to a specific day of the estrous cycle/early pregnancy. In conclusion, gene expression in endometria did not differ between pregnant and cycling heifers until Day 16 of pregnancy (i.e. the time of maternal recognition of pregnancy and production of interferon tau by conceptus trophectoderm); however, elevating P4 in early pregnancy programmed changes in gene expression in endometria that are hypothesized to impact early conceptus growth and development. Thus, on Days 5, 7 and 13 differential gene expression was affected by P4, but on Day 16 the conceptus primarily influenced gene expression in uterine endometria of heifers.
2011-10-18 | GSE33030 | GEO