Downregulated Luteolytic Pathways in Early Pregnant Bovine CL: Mimicry by Interferon-tau in vitro
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA685199 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13253951.fastq.gz | Fastqsanger.gz | |||
| SRR13253952.fastq.gz | Fastqsanger.gz | |||
| SRR13253953.fastq.gz | Fastqsanger.gz | |||
| SRR13253954.fastq.gz | Fastqsanger.gz | |||
| SRR13253955.fastq.gz | Fastqsanger.gz |
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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.
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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:Next-generation sequencing (RNA-Seq) was performed on CL from the late luteal phase and compared with normally luteolyzing CL collected at the prepartum P4 decrease, and following antigestagen (Aglepristone)- treatment. The analysis of transcriptomes presented herein supports the hypothesis describing luteal regression in non-pregnant dogs as a degenerative process devoid of the acute luteolytic principle and without an acute involvement of the immune system observed prepartum. The contribution of the immune system seems, however, to be critical in the PGF2alpha-mediated active prepartum luteolysis, which appears to be an acute immune process. The antigestagen-medited effects point towards the withdrawal of the luteotropic function of P4 with lesser involvement of immune system than during natural luteolysis. In summary, a deeper insights have been obtained into possible endocrine, paracrine and autocrine mechanisms governing the luteal life span in the domestic dog during pregnancy and in non-pregnant cycles.
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Project description:Embryo mortality (EM) contributes to infertility, but its exact mechanism is poorly understood. It was hypothesized that bovine EM pregnancies have impaired conceptus-derived interferon-tau (IFNT) release and action and are associated with altered transcriptome responses. The objective was to discover transcriptome response in EM tissues (endometrium [ENDO], corpus luteum [CL] and peripheral blood mononuclear cells [PBMC]) in lactating Holstein-Friesian cows. Two experiments (E1, E2) in day 16 pregnant (exposed to semen; E1:n=13, E2:n=15) or non-pregnant (NP; not exposed to semen; E1:n=7, E2:n=7) cows were completed. Uterine flushings (UF) and tissues were collected. Pregnant cows were also classified based on conceptus morphology and appearance for EM (E1:n=5, E2:n=6) or normal (N) conceptuses (E1:n=8, E2:n=9). Conceptuses and tissues were RNA sequenced and analyzed. The N conceptuses were longer (P<0.05) than EM conceptuses. The IFNT protein concentrations in UF were greater in N compared to EM and NP cows in E1 but not for E2. The ENDO conjugated ISG15 concentrations were greater in N (E1) than EM and NP cows but N (E2) were only greater than NP not EM cows. The major up-regulated canonical pathway in EM conceptuses was T helper 1 (Th1) and Th2. The ENDO had a massive increase in interferon stimulated genes in N and EM compared to NP cows. Estradiol-associated genes were up-regulated in EM compared to N ENDO for E2. The PBMC in E1 reflected up-regulation of genes associated Th1 immune responses in EM compared to N cows. Luteolytic pathways were upregulated in EM CL compared to N cows. This disruption of maternal recognition of pregnancy in EM pregnancies entails a massive T helper immune response within the conceptus, estradiol re-modelling of the ENDO, abnormal immune system in PBMC, luteolysis cascade in CL and, potentially, loss of pregnancy.
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Project description:Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is a key regulator of oxytocin. Our objectives were to measure serum concentrations of LNPEP and determine the presence of LNPEP and oxytocin (OXT) in equine tissue. LNPEP was measured by ELISA in serum obtained: every other day from ovulation (D0) until D21-22 in nonpregnant and pregnant mares; and at around 320 days of gestation, at 24 hours before foaling, and 20 min and 2 hrs post-foaling. Placenta from the pregnant horn (PH), nonpregnant horn (NPH) and body (B) and from tissues of 6 nonpregnant mares were homogenized and extracts obtained for protein determination and LNPEP ELISA assay. Identification of LNPEP and OXT protein in tissues was also performed via western blot (WB), immunohistochemistry (IHC) and liquid chromatography-mass spectrometry (LC-MS/MS). Furthermore, in situ hybridization was performed for LNPEP and OXT on endometrium, myometrium, pituitary and corpus luteum (CL). No difference in serum LNPEP concentration was identified between any of the groups. The NPH had the lowest quantities of LNPEP (P<0.05). The highest to lowest LNPEP U/mg protein by tissue were: myometrium> follicle wall> endometrium> kidney>CL> liver. Oxytocin was identified in the equine pituitary, CL and placenta. LNPEP may regulate the availability of luteal OXT locally.
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