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Reproduction (Cambridge, England)2018; 156(4); 313-330; doi: 10.1530/REP-18-0270

Dynamic changes in gene expression and signalling during trophoblast development in the horse.

Abstract: Equine chorionic girdle trophoblast cells play important endocrine and immune functions critical in supporting pregnancy. Very little is known about the genes and pathways that regulate chorionic girdle trophoblast development. Our aim was to identify genes and signalling pathways active in vivo in equine chorionic girdle trophoblast within a critical 7-days window. We exploited the late implantation of the equine conceptus to obtain trophoblast tissue. An Agilent equine 44K microarray was performed using RNA extracted from chorionic girdle and chorion (control) from equine pregnancy days 27, 30, 31 and 34 (n = 5), corresponding to the initiation of chorionic girdle trophoblast proliferation, differentiation and migration. Data were analysed using R packages limma and maSigPro, Ingenuity Pathway Analysis and DAVID and verified using qRT-PCR, promoter analysis, western blotting and migration assays. Microarray analysis showed gene expression (absolute log FC >2, FDR-adjusted P < 0.05) was rapidly and specifically induced in the chorionic girdle between days 27 and 34 (compared to day 27, day 30 = 116, day 31 = 317, day 34 = 781 genes). Pathway analysis identified 35 pathways modulated during chorionic girdle development (e.g. FGF, integrin, Rho GTPases, MAPK) including pathways that have limited description in mammalian trophoblast (e.g. IL-9, CD40 and CD28 signalling). Rho A and ERK/MAPK activity was confirmed as was a role for transcription factor ELF5 in regulation of the CGB promoter. The purity and accessibility of chorionic girdle trophoblast proved to be a powerful resource to identify candidate genes and pathways involved in early equine placental development.
Publication Date: 2018-10-01 PubMed ID: 29991567DOI: 10.1530/REP-18-0270Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research is designed to study the changes in gene expression and signaling pathways in equine chorionic girdle trophoblast cells during a crucial period of pregnancy in horses. These cells have key endocrine and immune functions that aid in sustaining pregnancy, but limited knowledge exists about the genes and pathways that direct their development.

Objective and Methodology

  • The purpose of the research was to identify active genes and signaling pathways in equine chorionic girdle trophoblast during a seven-day window critical for these cells’ development.
  • The investigators took advantage of the late implantation of the equine conceptus to gain trophoblast tissue.
  • The team conducted an Agilent equine 44K microarray, using RNA drawn from the chorionic girdle and the chorion (used as a control).
  • This study was carried out during days 27, 30, 31, and 34 of equine pregnancy, times corresponding to the chorionic girdle trophoblast cells’ initiation of proliferation, differentiation, and migration.

Analysis and Verification of Data

  • The data were analyzed using R packages limma and maSigPro, Ingenuity Pathway Analysis and DAVID.
  • Verification was done through qRT-PCR, promoter analysis, western blotting, and migration assays.

Results of the Study

  • Microarray analysis showed that gene expression was quickly and specifically induced in the chorionic girdle between days 27 and 34.
  • Pathway analysis identified 35 pathways that were modulated during chorionic girdle development, such as FGF, integrin, Rho GTPases, MAPK.
  • The analyses also showcased pathways that have received less description in mammalian trophoblast, such as IL-9, CD40 and CD28 signalling.
  • Rho A and ERK/MAPK activity was authenticated along with a role for transcription factor ELF5 in the regulation of the CGB promoter.

Conclusion

  • The purity and accessibility of chorionic girdle trophoblast proved to be an influential resource to identify the candidate genes and pathways involved in early equine placental development.

Cite This Article

APA
Read JE, Cabrera-Sharp V, Offord V, Mirczuk SM, Allen SP, Fowkes RC, de Mestre AM. (2018). Dynamic changes in gene expression and signalling during trophoblast development in the horse. Reproduction, 156(4), 313-330. https://doi.org/10.1530/REP-18-0270

Publication

ISSN: 1741-7899
NlmUniqueID: 100966036
Country: England
Language: English
Volume: 156
Issue: 4
Pages: 313-330
PII: REP-18-0270

Researcher Affiliations

Read, Jordan E
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK
Cabrera-Sharp, Victoria
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK
Offord, Victoria
  • Research Support Office, The Royal Veterinary College, London, UK
Mirczuk, Samantha M
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK
Allen, Steve P
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK
Fowkes, Robert C
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK
de Mestre, Amanda M
  • Department of Comparative Biomedical Sciences, The Royal Veterinary College, Hertfordshire, UK

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Citations

This article has been cited 1 times.
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