Transcriptional effects of prostaglandins inhibition in the early canine Corpus Luteum
Ontology highlight
ABSTRACT:
ORGANISM(S): Canis lupus familiaris
PROVIDER: GSE130369 | GEO | 2019/12/01
REPOSITORIES: GEO
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Project description:The downregulated pathways in early pregnant CL unraveled in this study span a wide range of luteal functions: angiogenesis, apoptosis and survival, steroidogenesis, and ECM remodeling. Notably, selective DEGs in luteinized granulosa cells (LGCs) were modulated by IFNT in vitro in a similar manner to their regulation in the CL of P cows, suggesting that IFNT may contribute to the characteristics of early pregnant CL.
2021-06-23 | GSE163212 | GEO
Project description:Purpose:We used transcriptome sequencing to characterize CL function during diestrus in non-pregnant female dogs. The aim of this study was to identify genes (and their respective pathways) that are differentially expressed (DE) during diestrus in order to obtain a broader understanding of CL function and developmental control. Methods:The CL were obtained from non-pregnant bitches (n=30) on days 10, 20, 30, 40, 50 and 60 (n=3/group) post-ovulation (p.o.). Sequencing was performed on an Illumina HiSeq 2000 using the paired-end reads protocol and the Illumina TruSeq PE cluster kit v3-cBotHS (Illumina). The reads were mapped against the reference genome (Canis_familiaris. CanFam 3.1.75.dna.toplevel.fa) using TopHat v2.0.9, and the transcripts were assembled using Cufflinks and identified by their Ensembl name. The relative abundance of transcripts of RNA-seq fragments was measured by Cufflinks in fragment per kilobase of exon model per million mapped reads (FPKM). We performed real-time PCR (qPCR) to validate the RNAseq results using Taqman assays. Results: A total of 771,208,718 reads (approximately 42 million per sample) was generated, with size of 100 bp. The software TopHat v2.0.9 assigned the reads to 34,408 genes, of which 9,000 were not annotated in the canine genome. 5,116 genes were differentially expressed (DEGs), although 1,106 of those were not yet annotated in the canine genome. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that many of the DE genes were related to cell proliferation, cell survival, angiogenesis, and immune system, which may regulate luteal formation and regression. Conclusion: This study provides fundamental data that could prove useful in reproductive research regarding the regulation of CL lifespan.
2019-07-01 | GSE89293 | GEO
Project description:Although rescue of the corpus luteum is required for pregnancy, luteal function during maternal recognition of pregnancy remains largely unexplored. CL were collected from pregnant cattle on days 14, 17, 20, and 23, to encompass the maternal recognition of pregnancy period. Nanostring technology was used to profile miRNA. A total of 27 miRNA changed. MiRNA that increased were predicted to inhibit phosphatidylinositol signaling, while those that decreased may be negative regulators of steroidogenesis. Overall, these data indicate that there are changes in the CL of pregnancy that are important for continued luteal function.
2020-03-28 | GSE147623 | GEO
Project description:Comparative genome wide gene expression profiles of small and large luteal cells from characterized mature CL are not currently available in any species. During present study, transcriptome differences of small and large luteal cells werte comprehensively analyzed to understand the specific functional roles of small and large luteal cells in mature bovine CL.
2018-03-31 | GSE106641 | GEO
Project description:The LH-like molecule chorionic gonadotropin (CG) is secreted by the macaque conceptus during and following implantation, M-bM-^@M-^\rescuingM-bM-^@M-^] the CL from impending regression and extending its functional lifespan in early pregnancy for approximately two weeks. CG binds to the same receptor as LH; i.e., LHCGR, and promotes production of steroids and other factors such as relaxin (RLN1). Our research group recently used AffymetrixM-bM-^DM-" rhesus macaque total genome arrays to compare the effects of CG on the luteal transcriptome from rhesus females during simulated early pregnancy (SEP) with changes during luteal regression in the non-fecund menstrual cycle. This analysis demonstrated that CG-rescue affected expression levels of 4,500 mRNA transcripts between days 10 and 15 of the luteal phase. Previous analyses indicated that a portion of the transcriptome in the macaque CL of the menstrual cycle was regulated indirectly by LH via the local actions of steroid hormones, including progesterone (P). Therefore, this study was designed to distinguish CG-regulated luteal genes that are dependent versus independent of local steroid (P) action. A protocol of increasing dosages of hCG (SEP) was begun on day 9 of the luteal phase in rhesus females combined with concurrent administration of the 3BHSD inhibitor trilostane (TRL) +/- the synthetic progestin (P) R5020. CL were collected on day 10 (no treatment) of the luteal phase to serve as a baseline comparison (n=8), 1 day of SEP (Day 10+hCG+/-TRL+/-R5020) and 6 days of SEP (Day 15+hCG+/-TRL+/-R5020); n=4/group. In the presence of CG, treatment with TRL reduced serum P levels to less than 1 ng/ ml after 1 day and all of the Day 15+h+TRL-treated females initiated menses before CL collection. The isolated CL were processed for total RNA and hybridized to microarrays. Compared to hCG treatment alone, 50 significantly altered mRNA transcripts were identified on day 10, rising to 95 on day 15 (P<0.05, M-bM-^IM-% 2-fold change in gene expression). The mRNA levels for several genes were validated in CL by real-time PCR. RNL1 levels increased with CG-treatment, but were not affected by steroid ablation, similar to previously reported relaxin protein expression. Steroid-sensitive genes included CDH11, IL1RN, INSL3, LDLR, OPA1, SERPINE1, SFRP4, and TNSF13B1; however differential sensitivity was observed and effects of steroid ablation and P replacement varied by day. Expression of some genes (e.g., 3BHSD2, ADAMTS1, ADAMTS5, MMP9, STAR, and VEGFA) previously identified as steroid regulated in the macaque CL during the menstrual cycle were not significantly altered by steroid ablation and P replacement during CG exposure in SEP. These data indicate that the majority of CG-regulated luteal transcripts are differentially expressed independently of local steroid actions. The proportion of steroid sensitive mRNA transcripts in the presence of CG is much smaller than in the presence of LH during the non-fecund cycle. Nevertheless, the steroid-regulated genes in the macaque CL may be essential during early pregnancy, based on the previous report that TRL treatment initiates premature structural regression of the CL during SEP. These data reinforce the concept that the structure, function, and regulation of the rescued CL in early pregnancy is different from the CL of the menstrual cycle. A protocol of increasing dosages of hCG (SEP) was begun on day 9 of the luteal phase in rhesus females combined with concurrent administration of the 3BHSD inhibitor trilostane (TRL) +/- the synthetic progestin (P) R5020. CL were collected on day 10 (no treatment) of the luteal phase to serve as a baseline comparison (n=8), 1 day of SEP (Day 10+hCG+/-TRL+/-R5020) and 6 days of SEP (Day 15+hCG+/-TRL+/-R5020); n=4/group.The isolated CL were processed for total RNA and hybridized to Affymetrix Rhesus Genome microarrays.
2012-12-01 | E-GEOD-29363 | biostudies-arrayexpress
Project description:The molecular and cellular processes required for development, function, and regression of the primate corpus luteum (CL) are poorly defined. We hypothesized that there are dynamic changes in gene expression occurring during the CL lifespan, which represent proteins and pathways critical to its regulation. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during the luteal phase of natural menstrual cycles. CL were collected between days 3-5 (early stage), 7-8 (mid), 10-12 (mid-late), 14-16 (late), or 18-19 (very-late) after the midcycle LH surge. From the early through very-late stages, 3234 transcripts were differentially expressed, with 879 occurring from the early through late stages that encompass the processes of luteinization, maintenance, and functional regression. To characterize gene changes most relevant to these processes, ontology analysis was performed using the list of 879 differentially expressed transcripts. Four main groups of related genes were identified with relevance to luteal physiology including: 1) immune function; 2) hormone and growth factor signaling; 3) steroidogenesis; and 4) prostaglandin biosynthesis, metabolism, and signaling. A subset of genes representing each of the four major categories was selected for validation of microarray results by quantitative real-time PCR. Results in mRNA levels were similar between the two methodologies for 17 of 18 genes. Additionally, protein levels for 3 genes were determined by Western blot analysis to parallel mRNA levels. This database will facilitate the identification of many novel or previously underappreciated pathways that regulate the structure and function of the primate CL. Keywords: time course The sole experimental factor is the stage of the luteal phase when the CL was collected. There were five stages: 1) between days 3-5 post LH-surge (early stage), 2) 7-8 (mid stage), 3) 10-12 (mid-late stage), 4) 14-16 (late stage), and 5) 18-19 (very-late stage). The early CL are undergoing luteinization, the mid are fully functional CL that are at their peak progesterone producing capacity, the mid-late stage is a transitionary period where CL are still producing significant quantities of progesterone but are nearing the time when luteolysis initiates in non-conception cycles, the late stage corresponds to CL undergoing functional regression (cessation of progesterone secretion), and the very-late stage (menses) is when the structural remodeling and apoptosis associated with luteolysis is occurring. Measuring mRNA levels in CL collected during these periods provides a comprehensive analysis of the primate transcriptome throughout CL lifespan. There were 20 biological replicates: n = 4 per stage, 5 stages in total. The early CL were used as the baseline reference for gene expression.
2008-02-08 | E-GEOD-10367 | biostudies-arrayexpress
Project description:The corpus luteum (CL) is essential for maintenance of pregnancy in all mammals and luteal rescue, which occurs around day 16-19 in the cow, is necessary to maintain luteal progesterone production. Transcriptomic and proteomic profiling were performed to compare the day 17 bovine CL of the estrous cycle and pregnancy. Among mRNA and proteins measured, 140 differentially abundant mRNA and 24 differentially abundant proteins were identified. Pathway analysis was performed using four programs. Modulated pathways included T cell receptor signaling, vascular stability, cytokine signaling, and extracellular matrix remodeling. Two mRNA that were less in pregnancy were regulated by prostaglandin (PG) F2A in culture, while two mRNA that were greater in pregnancy were regulated by interferon tau (IFNT). To identify mRNA that could be critical regulators of luteal fate, the mRNA that were differentially abundant during early pregnancy were compared to mRNA that were differentially abundant during luteal regression. Eight mRNA were common to both datasets, including mRNA related to regulation of steroidogenesis and gene transcription. A subset of differentially abundant mRNA and proteins, including those associated with extracellular matrix functions, were predicted targets of differentially abundant microRNA (miRNA). Integration of miRNA and protein data, using miRPath, revealed pathways such as extracellular matrix-receptor interactions, abundance of glutathione, and cellular metabolism and energy balance. Overall, this study has provided a comprehensive profile of molecular changes in the corpus luteum during maternal recognition of pregnancy and has indicated that some of these functions may be miRNA-regulated.
2019-08-04 | GSE135342 | GEO
Project description:The molecular and cellular processes required for development, function, and regression of the primate corpus luteum (CL) are poorly defined. We hypothesized that there are dynamic changes in gene expression occurring during the CL lifespan, which represent proteins and pathways critical to its regulation. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during the luteal phase of natural menstrual cycles. CL were collected between days 3-5 (early stage), 7-8 (mid), 10-12 (mid-late), 14-16 (late), or 18-19 (very-late) after the midcycle LH surge. From the early through very-late stages, 3234 transcripts were differentially expressed, with 879 occurring from the early through late stages that encompass the processes of luteinization, maintenance, and functional regression. To characterize gene changes most relevant to these processes, ontology analysis was performed using the list of 879 differentially expressed transcripts. Four main groups of related genes were identified with relevance to luteal physiology including: 1) immune function; 2) hormone and growth factor signaling; 3) steroidogenesis; and 4) prostaglandin biosynthesis, metabolism, and signaling. A subset of genes representing each of the four major categories was selected for validation of microarray results by quantitative real-time PCR. Results in mRNA levels were similar between the two methodologies for 17 of 18 genes. Additionally, protein levels for 3 genes were determined by Western blot analysis to parallel mRNA levels. This database will facilitate the identification of many novel or previously underappreciated pathways that regulate the structure and function of the primate CL. Keywords: time course
2008-02-08 | GSE10367 | GEO
Project description:The LH-like molecule chorionic gonadotropin (CG) is secreted by the macaque conceptus during and following implantation, “rescuing” the CL from impending regression and extending its functional lifespan in early pregnancy for approximately two weeks. CG binds to the same receptor as LH; i.e., LHCGR, and promotes production of steroids and other factors such as relaxin (RLN1). Our research group recently used Affymetrix™ rhesus macaque total genome arrays to compare the effects of CG on the luteal transcriptome from rhesus females during simulated early pregnancy (SEP) with changes during luteal regression in the non-fecund menstrual cycle. This analysis demonstrated that CG-rescue affected expression levels of 4,500 mRNA transcripts between days 10 and 15 of the luteal phase. Previous analyses indicated that a portion of the transcriptome in the macaque CL of the menstrual cycle was regulated indirectly by LH via the local actions of steroid hormones, including progesterone (P). Therefore, this study was designed to distinguish CG-regulated luteal genes that are dependent versus independent of local steroid (P) action. A protocol of increasing dosages of hCG (SEP) was begun on day 9 of the luteal phase in rhesus females combined with concurrent administration of the 3BHSD inhibitor trilostane (TRL) +/- the synthetic progestin (P) R5020. CL were collected on day 10 (no treatment) of the luteal phase to serve as a baseline comparison (n=8), 1 day of SEP (Day 10+hCG+/-TRL+/-R5020) and 6 days of SEP (Day 15+hCG+/-TRL+/-R5020); n=4/group. In the presence of CG, treatment with TRL reduced serum P levels to less than 1 ng/ ml after 1 day and all of the Day 15+h+TRL-treated females initiated menses before CL collection. The isolated CL were processed for total RNA and hybridized to microarrays. Compared to hCG treatment alone, 50 significantly altered mRNA transcripts were identified on day 10, rising to 95 on day 15 (P<0.05, ≥ 2-fold change in gene expression). The mRNA levels for several genes were validated in CL by real-time PCR. RNL1 levels increased with CG-treatment, but were not affected by steroid ablation, similar to previously reported relaxin protein expression. Steroid-sensitive genes included CDH11, IL1RN, INSL3, LDLR, OPA1, SERPINE1, SFRP4, and TNSF13B1; however differential sensitivity was observed and effects of steroid ablation and P replacement varied by day. Expression of some genes (e.g., 3BHSD2, ADAMTS1, ADAMTS5, MMP9, STAR, and VEGFA) previously identified as steroid regulated in the macaque CL during the menstrual cycle were not significantly altered by steroid ablation and P replacement during CG exposure in SEP. These data indicate that the majority of CG-regulated luteal transcripts are differentially expressed independently of local steroid actions. The proportion of steroid sensitive mRNA transcripts in the presence of CG is much smaller than in the presence of LH during the non-fecund cycle. Nevertheless, the steroid-regulated genes in the macaque CL may be essential during early pregnancy, based on the previous report that TRL treatment initiates premature structural regression of the CL during SEP. These data reinforce the concept that the structure, function, and regulation of the rescued CL in early pregnancy is different from the CL of the menstrual cycle.
2012-12-01 | GSE29363 | GEO
Project description:Prostaglandin F2 alpha (PGF2alpha) brings about regression of the bovine corpus luteum (CL). This luteolytic property of PGF2alpha is used in the beef and dairy cattle to synchronize estrus. A limitation of this protocol is an insensitivity of the early CL to luteolytic actions of PGF2alpha. The mechanisms underlying luteal sensitivity (LS) are poorly understood. The early CL has maximum number of PGF2alpha receptors; therefore differences in signaling events might be responsible for LS. Hence differential gene expression at two developmental stages, days 4 (D-4) and 10 (D-10) post estrus, might account for differences in signal transduction pathways associated with LS. This possibility was examined in these studies. Microarray analysis (n=3 per stage) identified 167 genes that were differentially expressed (p<0.05). These were categorized into genes involved in cell signaling, steroidogenesis and metabolism, protein degradation, transcription regulation and DNA biosynthesis, protein biosynthesis and modification, extracellular matrix and cytoskeletal proteins, antioxidant property, miscellaneous, and unknown functions. Real-time PCR confirmed the differential expression of 9 randomly selected genes, including protein kinase C inhibitor protein-1 (KCIP-1) and regulator of G-protein signaling 2 (RGS2), observed in microarray. Further, the in vivo effect of exogenous PGF2ï¡ (n=3 per stage) on selected genes that were found differentially expressed during this developmental transition was examined. PGF2alpha increased the expression of a guanine nucleotide binding protein beta polypeptide 1 (GNB1) in D-4 CL and Ca2+/calmodulin dependent kinase kinase 2 beta (Camkk2) in D-10 CL. Therefore, GNB1, Camkk2, KCIP-1, and RGS2 are candidate genes that might play significant role in acquisition of luteal sensitivity to PGF2alpha. Keywords: Direct comparison between Day-4 and Day-10 CL, Developmental type comparision The present experiment made a direct comparision of gene expression in early (Day-4) vs mid-phase CL (Day-10). Each experiment contained two technical relicates per slide. Total of three biological replicates were used ( 3 animals for Day-4 and 3 animals for Day-10). Two Day-4 samples were labelled Cy5 and one Day-10 with Cy5. Two Day-10 samples were labelled with Cy3 and one Day-4 with Cy3. The Day-4 sample was control for our experiments.
2010-05-05 | E-GEOD-10662 | biostudies-arrayexpress