The study of progesterone action in human myometrial explants

Study hypothesis: Myometrial explants represent a superior model compared with cell culture models for the study of human myometrial progesterone (P4) signalling in parturition. Study finding: Gene expression analysis showed myometrial explants closely resemble the in vivo condition and the anti-in...

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Main Authors: Georgiou, E.X., Lei, K., Lai, P.F., Yulia, A., Herbert, B.R., Castellanos, Marcos, May, Sean, Sooranna, S.R., Johnson, M.R.
Format: Article
Published: Oxford University Press 2016
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Online Access:https://eprints.nottingham.ac.uk/38989/
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author Georgiou, E.X.
Lei, K.
Lai, P.F.
Yulia, A.
Herbert, B.R.
Castellanos, Marcos
May, Sean
Sooranna, S.R.
Johnson, M.R.
author_facet Georgiou, E.X.
Lei, K.
Lai, P.F.
Yulia, A.
Herbert, B.R.
Castellanos, Marcos
May, Sean
Sooranna, S.R.
Johnson, M.R.
author_sort Georgiou, E.X.
building Nottingham Research Data Repository
collection Online Access
description Study hypothesis: Myometrial explants represent a superior model compared with cell culture models for the study of human myometrial progesterone (P4) signalling in parturition. Study finding: Gene expression analysis showed myometrial explants closely resemble the in vivo condition and the anti-inflammatory action of P4 is not lost with labour onset. What is known already: Circulating P4 levels decline before the onset of parturition in most animals, but not in humans. This has led to the suggestion that there is a functional withdrawal of P4 action at the myometrial level prior to labour onset. However, to date, no evidence of a loss of P4 function has been provided, with studies hampered by a lack of a physiologically relevant model. Study design, samples/materials, methods: Myometrial biopsies obtained at Caesarean section were dissected into explants after a portion was immediately snap frozen (t = 0). Microarray analysis was used to compare gene expression of t = 0 with paired (i) explants, (ii) passage 4 myometrial cell cultures or (iii) the hTERT myometrial cell line. Western blotting and chemokine/cytokine assays were used to study P4 signalling in myometrial explants. Main results and the role of chance: Gene expression comparison of t = 0 to the three models demonstrated that explants more closely resemble the in vivo status. At the protein level, explants maintain both P4 receptor (PR) and glucocorticoid receptor (GR) levels versus t = 0 whereas cells only maintain GR levels. Additionally, treatment with 1 μM P4 led to a reduction in interleukin-1 (IL-1) β-driven cyclooxygenase-2 in explants but not in cells. P4 signalling in explants was PR-mediated and associated with a repression of p65 and c-Jun phosphorylation. Furthermore, the anti-inflammatory action of P4 was maintained after labour onset. Limitations/reasons for caution: There is evidence of basal inflammation in the myometrial explant model. Wider implications of the findings: Myometrial explants constitute a novel model to study P4 signalling in the myometrium and can be used to further elucidate the mechanisms of P4 action in human labour. Large scale data: Data deposited at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=gvmpggkurbgxfqf&acc=GSE77830.
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spelling nottingham-389892020-05-04T17:50:09Z https://eprints.nottingham.ac.uk/38989/ The study of progesterone action in human myometrial explants Georgiou, E.X. Lei, K. Lai, P.F. Yulia, A. Herbert, B.R. Castellanos, Marcos May, Sean Sooranna, S.R. Johnson, M.R. Study hypothesis: Myometrial explants represent a superior model compared with cell culture models for the study of human myometrial progesterone (P4) signalling in parturition. Study finding: Gene expression analysis showed myometrial explants closely resemble the in vivo condition and the anti-inflammatory action of P4 is not lost with labour onset. What is known already: Circulating P4 levels decline before the onset of parturition in most animals, but not in humans. This has led to the suggestion that there is a functional withdrawal of P4 action at the myometrial level prior to labour onset. However, to date, no evidence of a loss of P4 function has been provided, with studies hampered by a lack of a physiologically relevant model. Study design, samples/materials, methods: Myometrial biopsies obtained at Caesarean section were dissected into explants after a portion was immediately snap frozen (t = 0). Microarray analysis was used to compare gene expression of t = 0 with paired (i) explants, (ii) passage 4 myometrial cell cultures or (iii) the hTERT myometrial cell line. Western blotting and chemokine/cytokine assays were used to study P4 signalling in myometrial explants. Main results and the role of chance: Gene expression comparison of t = 0 to the three models demonstrated that explants more closely resemble the in vivo status. At the protein level, explants maintain both P4 receptor (PR) and glucocorticoid receptor (GR) levels versus t = 0 whereas cells only maintain GR levels. Additionally, treatment with 1 μM P4 led to a reduction in interleukin-1 (IL-1) β-driven cyclooxygenase-2 in explants but not in cells. P4 signalling in explants was PR-mediated and associated with a repression of p65 and c-Jun phosphorylation. Furthermore, the anti-inflammatory action of P4 was maintained after labour onset. Limitations/reasons for caution: There is evidence of basal inflammation in the myometrial explant model. Wider implications of the findings: Myometrial explants constitute a novel model to study P4 signalling in the myometrium and can be used to further elucidate the mechanisms of P4 action in human labour. Large scale data: Data deposited at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=gvmpggkurbgxfqf&acc=GSE77830. Oxford University Press 2016-05-26 Article PeerReviewed Georgiou, E.X., Lei, K., Lai, P.F., Yulia, A., Herbert, B.R., Castellanos, Marcos, May, Sean, Sooranna, S.R. and Johnson, M.R. (2016) The study of progesterone action in human myometrial explants. Molecular Human Reproduction, 22 (8). pp. 877-889. ISSN 1460-2407 Progesterone Withdrawal Nuclear Receptors Myometrial Explants Smooth Muscle Parturition Myometrial Cells hTERT http://molehr.oxfordjournals.org/content/22/8/877 doi:10.1093/molehr/gaw037 doi:10.1093/molehr/gaw037
spellingShingle Progesterone Withdrawal
Nuclear Receptors
Myometrial Explants
Smooth Muscle
Parturition
Myometrial Cells
hTERT
Georgiou, E.X.
Lei, K.
Lai, P.F.
Yulia, A.
Herbert, B.R.
Castellanos, Marcos
May, Sean
Sooranna, S.R.
Johnson, M.R.
The study of progesterone action in human myometrial explants
title The study of progesterone action in human myometrial explants
title_full The study of progesterone action in human myometrial explants
title_fullStr The study of progesterone action in human myometrial explants
title_full_unstemmed The study of progesterone action in human myometrial explants
title_short The study of progesterone action in human myometrial explants
title_sort study of progesterone action in human myometrial explants
topic Progesterone Withdrawal
Nuclear Receptors
Myometrial Explants
Smooth Muscle
Parturition
Myometrial Cells
hTERT
url https://eprints.nottingham.ac.uk/38989/
https://eprints.nottingham.ac.uk/38989/
https://eprints.nottingham.ac.uk/38989/