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Myometrial immune cells contribute to term parturition, preterm labour and post-partum involution in mice.

ABSTRACT: This study aimed to determine the mechanism of uterine activation during labour, both term (TL) and preterm (PTL). We hypothesized that the peripheral leucocytes are recruited to uterine tissues by locally produced cytokines where they contribute to the initiation of parturition. Mouse uteri were collected (i) during gestation, TL and post-partum (PP), (ii) during PTL initiated by intrauterine infusion of LPS (125 ?g) or (iii) injection of the progesterone receptor antagonist RU486 and analysed for multiple cytokine expression levels by real-time polymerase chain reaction (RT-PCR) and 23-plex Cytokine assay or enzymatically dispersed for assessment of immune cell populations. Markers of myeloid cell differentiation (Gr1, Neu7/4 and F4/80) were evaluated by FACS to define tissue macrophages (Macs), monocytes (M) and neutrophils (N) and by immunohistochemistry to detect tissue Macs and N. Our results indicate that: (1) Macs were elevated in mouse myometrium before TL (P < 0.05) followed by an increase in M and N; these changes were accompanied by an increase in multiple pro-inflammatory cytokines/chemokines genes. The expression of corresponding proteins increased PP. (2) TL and RU486-PTL models showed similar gene/protein expression profiles, (3) LPS-PTL was characterized by strong pro-inflammatory response and massive influx of N in myometrial tissues showing a pattern different from TL and RU486-PTL, (4) The PP period appears similar in all three models, with elevated myometrial cytokine levels and high infiltration of immune cells. We concluded that leucocytes infiltrate myometrium around the time of parturition implicating their potential role in labour activation (both term and preterm) and major role in PP uterine involution.

SUBMITTER: Shynlova O

PROVIDER: S-EPMC3823139 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature

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Myometrial immune cells contribute to term parturition, preterm labour and post-partum involution in mice.

Shynlova Oksana O Nedd-Roderique Tamara T Li Yunqing Y Dorogin Anna A Lye Stephen J SJ

Journal of cellular and molecular medicine 20121204 1

This study aimed to determine the mechanism of uterine activation during labour, both term (TL) and preterm (PTL). We hypothesized that the peripheral leucocytes are recruited to uterine tissues by locally produced cytokines where they contribute to the initiation of parturition. Mouse uteri were collected (i) during gestation, TL and post-partum (PP), (ii) during PTL initiated by intrauterine infusion of LPS (125 μg) or (iii) injection of the progesterone receptor antagonist RU486 and analysed ...[more]

PMID: 23205502

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Project description:Comparison of gene expression in labouring and non-labouring myometrium. In this work, gene expression in 48 (22 labouring, 26 non-labouring) myometrial samples was measured using Illumina HT-12 v4.0 BeadChips. Biopsy samples of full thickness lower segment specimens of human myometrium were selected from the Edinburgh Reproductive Tissue Biobank (ERTBB, http://www.crh.ed.ac.uk/biobank/). All tissue samples were collected during Caesarean section from participant tissue donors with informed and written consent, according to the ethical approval and governance granted to the Edinburgh Reproductive Tissues BioBank by the West of Scotland Research Ethics Committee 4 (09/S0704/3). In order to maximise biological replicates on the array, all 22 labouring samples that fulfilled our inclusion criteria and that were stored in the ERTBB as of August 2012 were selected. In order to minimise inter-group variation in baseline characteristics, a second group of non-labouring samples (n=26) were selected by matching to labouring samples on maternal BMI, gestational age, parity and maternal age. We did not exclude smokers or women who had received medication to induce labour. We restricted the number of non-labouring samples to 26 so that all samples (n=48) could fit on three array chips. Forty-five of the samples had been transferred into RNAlater® (Life Technologies, Invitrogen, Carlsbad, CA, USA) immediately after collection and then stored at -80°C. Three labouring samples had not been transferred to RNAlater® before freezing but were also included in the array analysis in order to maximise the number of labouring samples. Total RNA was isolated using TRI Reagent® and the Qiagen RNeasy Lipid Tissue kit protocol (Qiagen, Valencia, CA, USA). The quantity and quality of RNA was assessed using a NanoDrop 1000 (Thermo Scientific, Wilmington, DE, USA). RNA quality was further assessed in the biotin-labelled samples by the Wellcome Trust Clinical Research Facility, Edinburgh using a Bioanalyzer 2100 (Agilent Technologies, Wilmington, DE, USA). To prepare samples for the microarray experiment, 450ng total RNA was amplified and biotin-labelled using the Illumina® TotalPrepTM RNA Amplification Kit (Ambion, Austin, TX, USA). Samples were split randomly over four Illumina HT-12 v4.0 BeadChips to minimise any effect of inter-chip variability. One sample was used per well. The chips were imaged using a BeadArray Reader and raw data was obtained with Illumina BeadStudio software. Raw data were preprocessed using the Lumi package in R version 2.14.1. The data was subjected to Robust Multichip Average (RMA) background correction before quantile normalisation to remove non-biological systematic variation.

Project description:Changes in cell phenotype are thought to occur through the expression of groups of co-regulated genes within topologically associated domains (TADs). In this paper, we allocate genes expressed within the myometrium of the human uterus during the onset of term labour into TADs. Transformation of the myometrial cells of the uterus into a contractile phenotype during term human labour is the result of a complex interaction of different epigenomic and genomic layers. Recent work suggests that the transcription factor (TF) RELA lies at the top of this regulatory network. Using deep RNA sequencing (RNAseq) analysis of myometrial samples (n = 16) obtained at term from women undergoing caesarean section prior to or after the onset of labour, we have identified evidence for how other gene expression regulatory elements interact with TFs in the labour phenotype transition. Gene set enrichment analysis of our RNAseq data identified three modules of enriched genes (M1, M2 and M3), which in gene ontology studies are linked to matrix degradation, smooth muscle and immune gene signatures, respectively. These genes were predominantly located within chromosomal TADs suggesting co-regulation of expression. Our transcriptomic analysis also identified significant differences in the expression of long non-coding RNAs (lncRNA), microRNAs (miRNA) and TFs that were predicted to target genes within the TADs. Additionally, network analysis revealed 15 new lncRNA (MCM3AP-AS1, TUG1, MIR29B2CHG, HCG18, LINC00963, KCNQ1OT1, NEAT1, HELLPAR, SNHG16, NUTM2B-AS1, MALAT1, PSMA3-AS1, GABPB1-AS1, NORAD and NKILA) and 4 miRNA (mir-145, mir-223, mir-let-7a and mir-132) as top gene hubs with three TFs (NFKB1, RELA and ESR1) as master regulators. Together, these factors are likely to be involved in co-regulatory networks driving a myometrial transformation to generate an estrogen-sensitive phenotype. We conclude that lncRNA and miRNA targeting the estrogen receptor 1 and nuclear factor kappa B pathways play a key role in the initiation of human labour. For the first time, we perform an integrative analysis to present a multi-level genomic signature made of mRNA, non-coding RNA and TFs in the myometrium for spontaneous term labour.

Dataset Information

Myometrial immune cells contribute to term parturition, preterm labour and post-partum involution in mice.

Publications

Myometrial immune cells contribute to term parturition, preterm labour and post-partum involution in mice.

Similar Datasets

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