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Home / Equine Research Bank / J Anim Sci / Growth factor modulation of equine trophoblast mitosis and p...
Journal of animal science2018; 97(2); 865-873; doi: 10.1093/jas/sky473

Growth factor modulation of equine trophoblast mitosis and prostaglandin gene expression.

Abstract:  To provide insight into maternal recognition of pregnancy control in equids, the mitogenic and developmental effects of endometrium-expressed growth factors (epidermal growth factor (EGF), fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1)) were examined in equine iTr cells, an equine trophectoderm cell line. Initial western blots revealed that HGF and IGF-1 stimulate phosphorylation of AKT serine/threonine kinase 1 (AKT1) and EGF, FGF2, or HGF resulted in phosphorylation of both extracellular signal-regulated kinase 1 (ERK1) and ERK2. Mitotic activity was stimulated (P < 0.05) by EGF, FGF2, and HGF. Chemical disruption of mitogen-activated protein kinase kinases 1 and 2 (MEK1/2) phosphorylation suppresses (P < 0.05) proliferation in control and growth factor treated cells demonstrating a dependence on ERK1/2 for mitotic activity. Treatment of iTr cells with EGF or HGF in the presence of an AKT1 inhibitor prohibits (P < 0.05) growth factor stimulated proliferation. The effect of EGF, FGF2, HGF, and IGF-1 on steroid biosynthetic enzyme gene expression, including prostaglandin-endoperoxide synthase 2 (PTGS2), was determined by real-time PCR. Neither EGF, FGF2, nor IGF-1 affected PTGS2 expression while HGF caused a two-fold increase (P < 0.05) in expression. Co-supplementation with HGF and an AKT1 inhibitor did not block PTGS2 expression, whereas providing an MEK1/2 inhibitor prevented (P < 0.05) the HGF-mediated increase in PTGS2. These results provide novel evidence of a role for HGF in equine trophectoderm proliferation and prostaglandin biosynthesis.
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Publication Date: 2018-12-12 PubMed ID: 30535412PubMed Central: PMC6358241DOI: 10.1093/jas/sky473Google 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 focuses on how certain growth factors influence cell division and gene expression in the cells that form the outer layer of a very early embryo (trophectoderm) in horses. The researchers found these growth factors may play key roles in the mother’s recognition of pregnancy.

Investigating Growth Factors

  • The researchers studied the effects of four growth factors expressed by the endometrium (the lining of the uterus), namely epidermal growth factor (EGF), fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1), on an equine trophectoderm cell line, known as iTr cells. Initial protein tests showed that HGF and IGF-1 stimulate the activation of a protein called AKT serine/threonine kinase 1 (AKT1) while EGF, FGF2, or HGF resulted in the activation of two other proteins, extracellular signal-regulated kinase 1 (ERK1) and ERK2.

Growth Factors and Cell Division

  • Exposure to EGF, FGF2, and HGF significantly increased cell division activity, which was established through chemical disruption of MEK1/2, proteins that regulate cell division. The disruption also showed these growth factors depend on ERK1/2 for mitotic activity. The study further revealed that blocking AKT1 restricts the proliferative effect of EGF or HGF on these cells.

Growth Factors and Gene Expression

  • The study found varying effects of these growth factors on the expression of genes involved in steroid biosynthesis, including a specific enzyme, prostaglandin-endoperoxide synthase 2 (PTGS2). While EGF, FGF2, and IGF-1 did not alter PTGS2 expression, HGF caused a significant increase in its expression. The research also found that when an AKT1 inhibitor was used alongside HGF, it didn’t block PTGS2 expression. However, using an MEK1/2 inhibitor prevented the HGF-driven increase in PTGS2.

Conclusion

  • The results of this study provide new evidence that HGF may play an important role in the proliferation of the equine trophectoderm and in prostaglandin biosynthesis, all of which are vital processes for the early stages of pregnancy in horses.

Cite This Article

APA
Bonometti S, Menarim BC, Reinholt BM, Ealy AD, Johnson SE. (2018). Growth factor modulation of equine trophoblast mitosis and prostaglandin gene expression. J Anim Sci, 97(2), 865-873. https://doi.org/10.1093/jas/sky473

Publication

Journal of animal science
ISSN: 1525-3163
NlmUniqueID: 8003002
Country: United States
Language: English
Volume: 97
Issue: 2
Pages: 865-873

Researcher Affiliations

Bonometti, Susana
  • Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg.
Menarim, Bruno Carvalho
  • Department of Large Animal Clinical Sciences, Virginia Polytechnic Institute and State University, Blacksburg.
Reinholt, Brad M
  • Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg.
Ealy, Alan D
  • Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg.
Johnson, Sally E
  • Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg.

MeSH Terms

  • Animals
  • Cell Line
  • Cell Proliferation / drug effects
  • Endometrium / metabolism
  • Epidermal Growth Factor / pharmacology
  • Female
  • Fibroblast Growth Factor 2 / pharmacology
  • Hepatocyte Growth Factor / pharmacology
  • Horses / genetics
  • Horses / physiology
  • Insulin-Like Growth Factor I / pharmacology
  • Mitosis / drug effects
  • Phosphorylation / drug effects
  • Pregnancy
  • Prostaglandins / biosynthesis
  • Prostaglandins / genetics
  • Trophoblasts / cytology

References

This article includes 32 references
  1. Ababneh MM, Troedsson MH, Michelson JR, Seguin BE. Partial characterization of an equine conceptus prostaglandin inhibitory factor.. J Reprod Fertil Suppl 2000;(56):607-13.
    pubmed: 20681176
  2. Allen WR, Gower S, Wilsher S. Localisation of epidermal growth factor (EGF), its specific receptor (EGF-R) and aromatase at the materno-fetal interface during placentation in the pregnant mare.. Placenta 2017 Feb;50:53-59.
    doi: 10.1016/j.placenta.2016.12.024pubmed: 28161062google scholar: lookup
  3. Berglund LA, Sharp DC, Vernon MW, Thatcher WW. Effect of pregnancy and collection technique on prostaglandin F in the uterine lumen of Pony mares.. J Reprod Fertil Suppl 1982;32:335-41.
    pubmed: 6962869
  4. Betteridge KJ, Eaglesome MD, Mitchell D, Flood PF, Beriault R. Development of horse embryos up to twenty two days after ovulation: observations on fresh specimens.. J Anat 1982 Aug;135(Pt 1):191-209.
    pmc: PMC1168142pubmed: 7130052
  5. Boerboom D, Brown KA, Vaillancourt D, Poitras P, Goff AK, Watanabe K, Doré M, Sirois J. Expression of key prostaglandin synthases in equine endometrium during late diestrus and early pregnancy.. Biol Reprod 2004 Feb;70(2):391-9.
    doi: 10.1095/biolreprod.103.020800pubmed: 14561653google scholar: lookup
  6. Bonilla AQ, Ozawa M, Hansen PJ. Timing and dependence upon mitogen-activated protein kinase signaling for pro-developmental actions of insulin-like growth factor 1 on the preimplantation bovine embryo.. Growth Horm IGF Res 2011 Apr;21(2):107-11.
    doi: 10.1016/j.ghir.2011.03.003pubmed: 21459028google scholar: lookup
  7. Cooke FN, Pennington KA, Yang Q, Ealy AD. Several fibroblast growth factors are expressed during pre-attachment bovine conceptus development and regulate interferon-tau expression from trophectoderm.. Reproduction 2009 Feb;137(2):259-69.
    doi: 10.1530/REP-08-0396pubmed: 18996977google scholar: lookup
  8. Dilly M, Hambruch N, Haeger JD, Pfarrer C. Epidermal growth factor (EGF) induces motility and upregulates MMP-9 and TIMP-1 in bovine trophoblast cells.. Mol Reprod Dev 2010 Jul;77(7):622-9.
    doi: 10.1002/mrd.21197pubmed: 20578063google scholar: lookup
  9. Ealy AD, Eroh ML, Sharp DC 3rd. Prostaglandin H synthase Type 2 is differentially expressed in endometrium based on pregnancy status in pony mares and responds to oxytocin and conceptus secretions in explant culture.. Anim Reprod Sci 2010 Jan;117(1-2):99-105.
    doi: 10.1016/j.anireprosci.2009.03.014pubmed: 19443143google scholar: lookup
  10. Fajardo-Puerta AB, Mato Prado M, Frampton AE, Jiao LR. Gene of the month: HGF.. J Clin Pathol 2016 Jul;69(7):575-9.
    doi: 10.1136/jclinpath-2015-203575pubmed: 27072686google scholar: lookup
  11. Gaivão MM, Rambags BP, Stout TA. Gastrulation and the establishment of the three germ layers in the early horse conceptus.. Theriogenology 2014 Jul 15;82(2):354-65.
    doi: 10.1016/j.theriogenology.2014.04.018pubmed: 24857628google scholar: lookup
  12. Heras-Sandoval D, Pérez-Rojas JM, Hernández-Damián J, Pedraza-Chaverri J. The role of PI3K/AKT/mTOR pathway in the modulation of autophagy and the clearance of protein aggregates in neurodegeneration.. Cell Signal 2014 Dec;26(12):2694-701.
    doi: 10.1016/j.cellsig.2014.08.019pubmed: 25173700google scholar: lookup
  13. Iqbal K, Chitwood JL, Meyers-Brown GA, Roser JF, Ross PJ. RNA-seq transcriptome profiling of equine inner cell mass and trophectoderm.. Biol Reprod 2014 Mar;90(3):61.
    doi: 10.1095/biolreprod.113.113928pmc: PMC4435230pubmed: 24478389google scholar: lookup
  14. Jeong W, Song G, Bazer FW, Kim J. Insulin-like growth factor I induces proliferation and migration of porcine trophectoderm cells through multiple cell signaling pathways, including protooncogenic protein kinase 1 and mitogen-activated protein kinase.. Mol Cell Endocrinol 2014 Mar 25;384(1-2):175-84.
    doi: 10.1016/j.mce.2014.01.023pubmed: 24508636google scholar: lookup
  15. Klein C. Novel equine conceptus?endometrial interactions on Day 16 of pregnancy based on RNA sequencing.. Reprod Fertil Dev 2015 May 5;.
    doi: 10.1071/RD14489pubmed: 25940503google scholar: lookup
  16. Laplante M, Sabatini DM. mTOR signaling in growth control and disease.. Cell 2012 Apr 13;149(2):274-93.
    doi: 10.1016/j.cell.2012.03.017pmc: PMC3331679pubmed: 22500797google scholar: lookup
  17. McDowell KJ, Sharp DC, Grubaugh W, Thatcher WW, Wilcox CJ. Restricted conceptus mobility results in failure of pregnancy maintenance in mares.. Biol Reprod 1988 Sep;39(2):340-8.
    doi: 10.1095/biolreprod39.2.340pubmed: 3179385google scholar: lookup
  18. Mesalam A, Lee KL, Khan I, Chowdhury MMR, Zhang S, Song SH, Joo MD, Lee JH, Jin JI, Kong IK. A combination of bovine serum albumin with insulin-transferrin-sodium selenite and/or epidermal growth factor as alternatives to fetal bovine serum in culture medium improves bovine embryo quality and trophoblast invasion by induction of matrix metalloproteinases.. Reprod Fertil Dev 2019 Jan;31(2):333-346.
    doi: 10.1071/RD18162pubmed: 30086822google scholar: lookup
  19. Michael DD, Alvarez IM, Ocón OM, Powell AM, Talbot NC, Johnson SE, Ealy AD. Fibroblast growth factor-2 is expressed by the bovine uterus and stimulates interferon-tau production in bovine trophectoderm.. Endocrinology 2006 Jul;147(7):3571-9.
    doi: 10.1210/en.2006-0234pubmed: 16574787google scholar: lookup
  20. Reinholt BM, Bradley JS, Jacobs RD, Ealy AD, Johnson SE. Tissue organization alters gene expression in equine induced trophectoderm cells.. Gen Comp Endocrinol 2017 Jun 1;247:174-182.
    doi: 10.1016/j.ygcen.2017.01.030pubmed: 28161437google scholar: lookup
  21. Rose BV, Firth M, Morris B, Roach JM, Wathes DC, Verheyen KLP, de Mestre AM. Descriptive study of current therapeutic practices, clinical reproductive findings and incidence of pregnancy loss in intensively managed thoroughbred mares.. Anim Reprod Sci 2018 Jan;188:74-84.
    doi: 10.1016/j.anireprosci.2017.11.011pubmed: 29146097google scholar: lookup
  22. de Ruijter-Villani M, van Boxtel PR, Stout TA. Fibroblast growth factor-2 expression in the preimplantation equine conceptus and endometrium of pregnant and cyclic mares.. Theriogenology 2013 Dec;80(9):979-89.
    doi: 10.1016/j.theriogenology.2013.07.024pubmed: 24035195google scholar: lookup
  23. de Ruijter-Villani M, Deelen C, Stout TA. Expression of leukaemia inhibitory factor at the conceptus?maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare.. Reprod Fertil Dev 2015 Apr 17;.
    doi: 10.1071/RD14334pubmed: 25881292google scholar: lookup
  24. Sessions-Bresnahan DR, Heuberger AL, Carnevale EM. Obesity in mares promotes uterine inflammation and alters embryo lipid fingerprints and homeostasis.. Biol Reprod 2018 Oct 1;99(4):761-772.
    doi: 10.1093/biolre/ioy107pubmed: 29741587google scholar: lookup
  25. Sharp DC, McDowell KJ, Weithenauer J, Thatcher WW. The continuum of events leading to maternal recognition of pregnancy in mares.. J Reprod Fertil Suppl 1989;37:101-7.
    pubmed: 2810225
  26. Silva LA, Klein C, Ealy AD, Sharp DC. Conceptus-mediated endometrial vascular changes during early pregnancy in mares: an anatomic, histomorphometric, and vascular endothelial growth factor receptor system immunolocalization and gene expression study.. Reproduction 2011 Oct;142(4):593-603.
    doi: 10.1530/REP-11-0149pubmed: 21757474google scholar: lookup
  27. Spencer TE, Burghardt RC, Johnson GA, Bazer FW. Conceptus signals for establishment and maintenance of pregnancy.. Anim Reprod Sci 2004 Jul;82-83:537-50.
    doi: 10.1016/j.anireprosci.2004.04.014pubmed: 15271478google scholar: lookup
  28. Stout TA, Allen WR. Role of prostaglandins in intrauterine migration of the equine conceptus.. Reproduction 2001 May;121(5):771-5.
    pubmed: 11427165
  29. Stout TA, Allen WR. Prostaglandin E(2) and F(2 alpha) production by equine conceptuses and concentrations in conceptus fluids and uterine flushings recovered from early pregnant and dioestrous mares.. Reproduction 2002 Feb;123(2):261-8.
    pubmed: 11866693
  30. Watson ED, Sertich PL. Prostaglandin production by horse embryos and the effect of co-culture of embryos with endometrium from pregnant mares.. J Reprod Fertil 1989 Sep;87(1):331-6.
    doi: 10.1530/jrf.0.0870331pubmed: 2621704google scholar: lookup
  31. Xie M, McCoski SR, Johnson SE, Rhoads ML, Ealy AD. Combinatorial effects of epidermal growth factor, fibroblast growth factor 2 and insulin-like growth factor 1 on trophoblast cell proliferation and embryogenesis in cattle.. Reprod Fertil Dev 2017 Feb;29(2):419-430.
    doi: 10.1071/RD15226pubmed: 26304178google scholar: lookup
  32. Yu JS, Cui W. Proliferation, survival and metabolism: the role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination.. Development 2016 Sep 1;143(17):3050-60.
    doi: 10.1242/dev.137075pubmed: 27578176google scholar: lookup

Citations

This article has been cited 4 times.
  1. Gastal GDA, Scarlet D, Melchert M, Ertl R, Aurich C. Epigenetic Changes in Equine Embryos after Short-Term Storage at Different Temperatures. Animals (Basel) 2021 May 6;11(5).
    doi: 10.3390/ani11051325pubmed: 34066466google scholar: lookup
  2. Llobat L. Pluripotency and Growth Factors in Early Embryonic Development of Mammals: A Comparative Approach. Vet Sci 2021 May 4;8(5).
    doi: 10.3390/vetsci8050078pubmed: 34064445google scholar: lookup
  3. Ma Y, Yu X, Li YX, Wang YL. HGF/c-Met signaling regulates early differentiation of placental trophoblast cells. J Reprod Dev 2021 Apr 21;67(2):89-97.
    doi: 10.1262/jrd.2020-107pubmed: 33455972google scholar: lookup
  4. Ji K, Chen Y, Pan X, Chen L, Wang X, Wen B, Bao J, Zhong J, Lv Z, Zheng Z, Liu H. Single-cell and spatial transcriptomics reveal alterations in trophoblasts at invasion sites and disturbed myometrial immune microenvironment in placenta accreta spectrum disorders. Biomark Res 2024 Jun 3;12(1):55.
    doi: 10.1186/s40364-024-00598-6pubmed: 38831319google scholar: lookup
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