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Scientific reports2018; 8(1); 5249; doi: 10.1038/s41598-018-23537-6

Proteins involved in embryo-maternal interaction around the signalling of maternal recognition of pregnancy in the horse.

Abstract: During maternal recognition of pregnancy (MRP), a conceptus-derived signal leads to the persistence of the corpus luteum and the maintenance of gestation. In the horse, the nature of this signal remains to be elucidated. Several studies have focused on the changes in gene expression during MRP, but little information exists at the protein level. The aim of this study was to identify the proteins at the embryo-maternal interface around signalling of MRP in the horse (day 13) by means of mass spectrometry. A distinct influence of pregnancy was established, with 119 proteins differentially expressed in the uterine fluid of pregnant mares compared to cyclic mares and with upregulation of several inhibitors of the prostaglandin synthesis during pregnancy. By creating an overview of the proteins at the embryo-maternal interface in the horse, this study provides a solid foundation for further targeted studies of proteins potentially involved in embryo-maternal interactions, MRP and pregnancy loss in the horse.
Publication Date: 2018-03-27 PubMed ID: 29588480PubMed Central: PMC5869742DOI: 10.1038/s41598-018-23537-6Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 researchers conducted a study on horses to identify proteins involved in the embryo-maternal interaction, specifically during the maternal recognition of pregnancy (MRP). Using mass spectrometry, they found that pregnancy influenced the expression of 119 proteins in the uterine fluid of pregnant mares, and several inhibitors of prostaglandin synthesis were upregulated during pregnancy.

Objective of Research

  • The primary objective of this study was to identify and examine the proteins involved in the interaction between an embryo and its mother around the signalling of maternal recognition of pregnancy (MRP) in horses. Identifying these proteins would provide significant insights into the mechanisms of MRP and offer a foundation for further research into pregnancy loss in horses.

Methodology

  • To identify these proteins, the research team employed a method called mass spectrometry. They chose this technique as it is capable of identifying and quantifying individual proteins in a mixture based on their distinctive mass.
  • The study focused on day 13 of the MRP, a critical period when the conceptus-derived signal leads to the persistence of the corpus luteum, which is necessary for the maintenance of gestation in horses.
  • This research compared the proteins found in the uterine fluid of pregnant mares and cyclic mares to identify any differences.

Findings

  • The study discovered a significant influence of pregnancy on protein expression, with 119 proteins differentially expressed in the uterine fluid of pregnant mares compared to cyclic mares. This finding indicates that there are specific proteins associated with pregnancy and potentially with the process of maternal recognition of pregnancy.
  • Furthermore, the research observed an upregulation of various inhibitors of prostaglandin synthesis during pregnancy. As prostaglandins play a crucial role in various reproductive processes, including the regulation of the menstrual cycle and the induction of labor, an increased presence of these inhibitors during pregnancy could have significant implications.

Implications

  • The results provide a comprehensive overview of the proteins present at the embryo-maternal interface in horses, particularly during the signal of MRP. This knowledge could form the basis for further targeted studies into these proteins and their potential role in embryo-maternal interactions and pregnancy loss in horses.
  • The study’s discoveries surrounding the upregulation of inhibitors of prostaglandin synthesis also open up new avenues for research into the regulation of prostaglandin during pregnancy.

Cite This Article

APA
Smits K, Willems S, Van Steendam K, Van De Velde M, De Lange V, Ververs C, Roels K, Govaere J, Van Nieuwerburgh F, Peelman L, Deforce D, Van Soom A. (2018). Proteins involved in embryo-maternal interaction around the signalling of maternal recognition of pregnancy in the horse. Sci Rep, 8(1), 5249. https://doi.org/10.1038/s41598-018-23537-6

Publication

ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 8
Issue: 1
Pages: 5249

Researcher Affiliations

Smits, Katrien
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium. Katrien.Smits@UGent.be.
Willems, Sander
  • Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Gent, Belgium.
Van Steendam, Katleen
  • Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Gent, Belgium.
Van De Velde, Margot
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
De Lange, Valérie
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Ververs, Cyrillus
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Roels, Kim
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Govaere, Jan
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Van Nieuwerburgh, Filip
  • Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Gent, Belgium.
Peelman, Luc
  • Laboratory of Animal Genetics, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Deforce, Dieter
  • Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Gent, Belgium.
Van Soom, Ann
  • Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

MeSH Terms

  • Animals
  • Embryo, Mammalian / embryology
  • Embryo, Mammalian / metabolism
  • Female
  • Horses / embryology
  • Horses / metabolism
  • Pregnancy
  • Protein Interaction Maps
  • Proteins / metabolism
  • Signal Transduction
  • Uterus / metabolism

Conflict of Interest Statement

The authors declare no competing interests.

References

This article includes 68 references
  1. Short R. Implantation and the maternal recognition of pregnancy.. Foetal Autonomy 1969;2:31.
  2. Kindahl H, Knudsen O, Madej A, Edqvist LE. Progesterone, prostaglandin F-2 alpha, PMSG and oestrone sulphate during early pregnancy in the mare.. J Reprod Fertil Suppl 1982;32:353–359.
    pubmed: 6300390
  3. Bazer FW, Thatcher W. Theory of maternal recognition of pregnancy in swine based on estrogen controlled endocrine versus exocrine secretion of prostaglandin F 2α by the uterine endometrium.. Prostaglandins 1977;14:397–401.
    doi: 10.1016/0090-6980(77)90185-Xpubmed: 897228google scholar: lookup
  4. Lamming GE. Local action of trophoblast interferons in suppression of the development of oxytocin and oestradiol receptors in ovine endometrium.. J Reprod Fertil 1995;105:165–175.
    doi: 10.1530/jrf.0.1050165pubmed: 7490709google scholar: lookup
  5. Lamming GE, Mann GE. Control of endometrial oxytocin receptors and prostaglandin F2 alpha production in cows by progesterone and oestradiol.. J Reprod Fertil 1995;103:69–73.
    doi: 10.1530/jrf.0.1030069pubmed: 7707303google scholar: lookup
  6. Allen WR. Fetomaternal interactions and influences during equine pregnancy.. Reproduction 2001;121:513–527.
    doi: 10.1530/rep.0.1210513pubmed: 11277870google scholar: lookup
  7. Klein C, Troedsson MH. Maternal recognition of pregnancy in the horse: a mystery still to be solved.. Reprod Fertil Dev 2011;23:952–963.
    doi: 10.1071/RD10294pubmed: 22127001google scholar: lookup
  8. Vanderwall DK, Silvia WJ, Fitzgerald BP. Concentrations of oxytocin in the intercavernous sinus of mares during luteolysis: temporal relationship with concentrations of 13, 14-dihydro-15-keto-prostaglandin F2 alpha.. J Reprod Fertil 1998;112:337–346.
    doi: 10.1530/jrf.0.1120337pubmed: 9640273google scholar: lookup
  9. Goff AK, Pontbriand D, Sirois J. Oxytocin stimulation of plasma 15-keto-13, 14-dihydro prostaglandin F-2 alpha during the oestrous cycle and early pregnancy in the mare.. J Reprod Fertil Suppl 1987;35:253–260.
    pubmed: 3479581
  10. Starbuck GR, Stout TA, Lamming GE, Allen WR, Flint AP. Endometrial oxytocin receptor and uterine prostaglandin secretion in mares during the oestrous cycle and early pregnancy.. J Reprod Fertil 1998;113:173–179.
    doi: 10.1530/jrf.0.1130173pubmed: 9861156google scholar: lookup
  11. Boerboom D. Expression of key prostaglandin synthases in equine endometrium during late diestrus and early pregnancy.. Biol Reprod 2004;70:391–399.
    doi: 10.1095/biolreprod.103.020800pubmed: 14561653google scholar: lookup
  12. Ealy AD, Eroh ML, Sharp DC. 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;117:99–105.
  13. de Ruijter-Villani M, van Tol HT, Stout TA. Effect of pregnancy on endometrial expression of luteolytic pathway components in the mare.. Reprod Fertil Dev 2015;27:834–845.
    doi: 10.1071/RD13381pubmed: 24679480google scholar: lookup
  14. Wilsher S, Clutton-Brock A, Allen WR. Successful transfer of day 10 horse embryos: influence of donor-recipient asynchrony on embryo development.. Reproduction 2010;139:575–585.
    doi: 10.1530/REP-09-0306pubmed: 19948839google scholar: lookup
  15. Klein C, Scoggin KE, Ealy AD, Troedsson MH. Transcriptional profiling of equine endometrium during the time of maternal recognition of pregnancy.. Biol Reprod 2010;83:102–113.
    doi: 10.1095/biolreprod.109.081612pubmed: 20335638google scholar: lookup
  16. Klein C, Troedsson MH. Transcriptional profiling of equine conceptuses reveals new aspects of embryo-maternal communication in the horse.. Biol Reprod 2011;84:872–885.
    doi: 10.1095/biolreprod.110.088732pubmed: 21209420google scholar: lookup
  17. Merkl M. Microarray analysis of equine endometrium at days 8 and 12 of pregnancy.. Biol Reprod 2010;83:874–886.
    doi: 10.1095/biolreprod.110.085233pubmed: 20631402google scholar: lookup
  18. Bauersachs S, Wolf E. Transcriptome analyses of bovine, porcine and equine endometrium during the pre-implantation phase.. Anim Reprod Sci 2012;134:84–94.
  19. Klein C. Novel equine conceptus?endometrial interactions on Day 16 of pregnancy based on RNA sequencing.. Reprod Fertil Dev 2015.
    pubmed: 25940503
  20. Wright PC, Noirel J, Ow SY, Fazeli A. A review of current proteomics technologies with a survey on their widespread use in reproductive biology investigations.. Theriogenology 2012;77:738–765 e752.
  21. Vogel C, Marcotte EM. Insights into the regulation of protein abundance from proteomic and transcriptomic analyses.. Nature reviews. Genetics 2012;13:227–232.
    doi: 10.1038/nrg3185pmc: PMC3654667pubmed: 22411467google scholar: lookup
  22. Distler U, Kuharev J, Tenzer S. Biomedical applications of ion mobility-enhanced data-independent acquisition-based label-free quantitative proteomics.. Expert review of proteomics 2014;11:675–684.
    doi: 10.1586/14789450.2014.971114pubmed: 25327648google scholar: lookup
  23. Kayser JP, Kim JG, Cerny RL, Vallet JL. Global characterization of porcine intrauterine proteins during early pregnancy.. Reproduction 2006;131:379–388.
    doi: 10.1530/rep.1.00882pubmed: 16452731google scholar: lookup
  24. Jalali BM, Bogacki M, Dietrich M, Likszo P, Wasielak M. Proteomic analysis of porcine endometrial tissue during peri-implantation period reveals altered protein abundance.. Journal of proteomics 2015;125:76–88.
    doi: 10.1016/j.jprot.2015.05.003pubmed: 25976747google scholar: lookup
  25. Brooks K, Burns GW, Moraes JG, Spencer TE. Analysis of the Uterine Epithelial and Conceptus Transcriptome and Luminal Fluid Proteome During the Peri-Implantation Period of Pregnancy in Sheep.. Biol Reprod 2016;95(88):1–17.
    pubmed: 27535962
  26. Munoz M. Proteome of the early embryo-maternal dialogue in the cattle uterus.. J Proteome Res 2012;11:751–766.
    doi: 10.1021/pr200969apubmed: 22148898google scholar: lookup
  27. Forde N, Bazer FW, Spencer TE, Lonergan P. ‘Conceptualizing’ the Endometrium: Identification of Conceptus-Derived Proteins During Early Pregnancy in Cattle.. Biol Reprod 2015;92(156):1–13.
    pmc: PMC4652614pubmed: 25947061
  28. Forde N. Proteomic analysis of uterine fluid during the pre-implantation period of pregnancy in cattle.. Reproduction .
    pubmed: 24478148doi: 10.1530/rep-13-0010google scholar: lookup
  29. Tachibana Y. Expression of endometrial immune-related genes possibly functioning during early pregnancy in the mare.. J Reprod Dev 2013;59:85–91.
    pmc: PMC3943239pubmed: 23138119
  30. Klein C, Scoggin KE, Troedsson MH. The expression of interferon-stimulated gene 15 in equine endometrium.. Reprod Domest Anim 2011;46:692–698.
  31. Hartt LS. Temporal and spatial associations of oestrogen receptor alpha and progesterone receptor in the endometrium of cyclic and early pregnant mares.. Reproduction 2005;130:241–250.
    doi: 10.1530/rep.1.00596pubmed: 16049162google scholar: lookup
  32. Watson ED, Buckingham J, Bjorksten T, Nikolakopoulos E. Immunolocalization of oxytocin and neurophysin in the mare uterus.. J Reprod Fertil Suppl 2000;289–296.
    pubmed: 20681140
  33. Wolf CA, Maslchitzky E, Gregory RM, Jobim MI, Mattos RC. Effect of corticotherapy on proteomics of endometrial fluid from mares susceptible to persistent postbreeding endometritis.. Theriogenology 2012;77:1351–1359.
  34. Swegen A. From Peptide Masses to Pregnancy Maintenance: A Comprehensive Proteomic Analysis of The Early Equine Embryo Secretome, Blastocoel Fluid, and Capsule.. Proteomics 2017;17.
    pubmed: 28782881doi: 10.1002/pmic.201600433google scholar: lookup
  35. Distler U. Drift time-specific collision energies enable deep-coverage data-independent acquisition proteomics.. Nature methods 2014;11:167–170.
    doi: 10.1038/nmeth.2767pubmed: 24336358google scholar: lookup
  36. Hayes JD, Flanagan JU, Jowsey IR. Glutathione transferases.. Annual review of pharmacology and toxicology 2005;45:51–88.
  37. Luo L. Recombinant protein glutathione S-transferases P1 attenuates inflammation in mice.. Molecular immunology 2009;46:848–857.
    doi: 10.1016/j.molimm.2008.09.010pubmed: 18962899google scholar: lookup
  38. Wallner BP. Cloning and expression of human lipocortin, a phospholipase A2 inhibitor with potential anti-inflammatory activity.. Nature 1986;320:77–81.
    doi: 10.1038/320077a0pubmed: 2936963google scholar: lookup
  39. Huang KS. Two human 35 kd inhibitors of phospholipase A2 are related to substrates of pp60v-src and of the epidermal growth factor receptor/kinase.. Cell 1986;46:191–199.
    doi: 10.1016/0092-8674(86)90736-1pubmed: 3013422google scholar: lookup
  40. Smits K. The Equine Embryo Influences Immune-Related Gene Expression in the Oviduct.. Biol Reprod 2016;94:36.
    doi: 10.1095/biolreprod.115.136432pubmed: 26740593google scholar: lookup
  41. Ababneh MM, Troedsson MH. Endometrial phospholipase A2 activity during the oestrous cycle and early pregnancy in mares.. Reprod Domest Anim 2013;48:46–52.
  42. Albrecht D, Kniemeyer O, Brakhage AA, Guthke R. Missing values in gel-based proteomics.. Proteomics 2010;10:1202–1211.
    doi: 10.1002/pmic.200800576pubmed: 20077407google scholar: lookup
  43. Webb-Robertson BJ. Review, evaluation, and discussion of the challenges of missing value imputation for mass spectrometry-based label-free global proteomics.. J Proteome Res 2015;14:1993–2001.
    doi: 10.1021/pr501138hpmc: PMC4776766pubmed: 25855118google scholar: lookup
  44. Park ES. Phospholipase C-delta1 and oxytocin receptor signalling: evidence of its role as an effector.. Biochem J 1998;331(Pt 1):283–289.
    doi: 10.1042/bj3310283pmc: PMC1219350pubmed: 9512491google scholar: lookup
  45. Bu H. ERBP, a novel estrogen receptor binding protein enhancing the activity of estrogen receptor.. Biochem Biophys Res Commun 2004;317:54–59.
    doi: 10.1016/j.bbrc.2004.02.179pubmed: 15047147google scholar: lookup
  46. Montt-Guevara MM. Androgens Regulate T47D Cells Motility and Invasion through Actin Cytoskeleton Remodeling.. Frontiers in endocrinology 2016;7:136.
    doi: 10.3389/fendo.2016.00136pmc: PMC5043384pubmed: 27746764google scholar: lookup
  47. Song J. Expression and clinicopathological significance of oestrogen-responsive ezrin-radixin-moesin-binding phosphoprotein 50 in breast cancer.. Histopathology 2007;51:40–53.
  48. Tranguch S, Smith DF, Dey SK. Progesterone receptor requires a co-chaperone for signalling in uterine biology and implantation.. Reprod Biomed Online 2006;13:651–660.
    doi: 10.1016/S1472-6483(10)60655-4pubmed: 17169175google scholar: lookup
  49. Tranguch S. Cochaperone immunophilin FKBP52 is critical to uterine receptivity for embryo implantation.. Proceedings of the National Academy of Sciences of the United States of America 2005;102:14326–14331.
    doi: 10.1073/pnas.0505775102pmc: PMC1242310pubmed: 16176985google scholar: lookup
  50. Chen HY. Expression of FK506-binding protein 52 (FKBP52) in chorionic villi with early recurrent spontaneous abortion.. J Matern Fetal Neonatal Med 2015;28:1165–1169.
    doi: 10.3109/14767058.2014.947572pubmed: 25053194google scholar: lookup
  51. Crossett B, Allen WR, Stewart F. A 19 kDa protein secreted by the endometrium of the mare is a novel member of the lipocalin family.. Biochem J 1996;320(Pt 1):137–143.
    doi: 10.1042/bj3200137pmc: PMC1217908pubmed: 8947478google scholar: lookup
  52. Ellenberger C. Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare’s uterus and placenta throughout pregnancy.. Theriogenology 2008;70:746–757.
  53. Suire S, Stewart F, Beauchamp J, Kennedy MW. Uterocalin, a lipocalin provisioning the preattachment equine conceptus: fatty acid and retinol binding properties, and structural characterization.. Biochem J 2001;356:369–376.
    doi: 10.1042/bj3560369pmc: PMC1221847pubmed: 11368763google scholar: lookup
  54. Quinn BA, Hayes MA, Waelchli RO, Kennedy MW, Betteridge KJ. Changes in major proteins in the embryonic capsule during immobilization (fixation) of the conceptus in the third week of pregnancy in the mare.. Reproduction 2007;134:161–170.
    doi: 10.1530/REP-06-0241pubmed: 17641098google scholar: lookup
  55. Stewart F, Charleston B, Crossett B, Barker PJ, Allen WR. A novel uterine protein that associates with the embryonic capsule in equids.. J Reprod Fertil 1995;105:65–70.
    doi: 10.1530/jrf.0.1050065pubmed: 7490716google scholar: lookup
  56. Smits K. Influence of the uterine environment on the development of in vitro-produced equine embryos.. Reproduction 2012;143:173–181.
    pubmed: 22089531doi: 10.1530/rep-11-0217google scholar: lookup
  57. Schmidt A, Forne I, Imhof A. Bioinformatic analysis of proteomics data.. BMC systems biology 2014;8(Suppl 2):S3.
    doi: 10.1186/1752-0509-8-S2-S3pmc: PMC4108846pubmed: 25033288google scholar: lookup
  58. Vaudel M, Sickmann A, Martens L. Introduction to opportunities and pitfalls in functional mass spectrometry based proteomics.. Biochim Biophys Acta 2014;1844:12–20.
    doi: 10.1016/j.bbapap.2013.06.019pubmed: 23845992google scholar: lookup
  59. Piwowar AM, Lockyer NP, Vickerman JC. Salt effects on ion formation in desorption mass spectrometry: an investigation into the role of alkali chlorides on peak suppression in time-of-flight-secondary ion mass spectrometry.. Anal Chem 2009;81:1040–1048.
    doi: 10.1021/ac8020888pubmed: 19125566google scholar: lookup
  60. Noble WS. Mass spectrometrists should search only for peptides they care about.. Nature methods 2015;12:605–608.
    doi: 10.1038/nmeth.3450pmc: PMC4711994pubmed: 26125591google scholar: lookup
  61. Noble WS, Keich U. Mass spectrometrists should search for all peptides, but assess only the ones they care about Reply.. Nature methods 2017;14:644–644.
    doi: 10.1038/nmeth.4339pmc: PMC5740876pubmed: 28661496google scholar: lookup
  62. Sticker A, Martens L, Clement L. Mass spectrometrists should search for all peptides, but assess only the ones they care about.. Nature methods 2017;14:643–644.
    doi: 10.1038/nmeth.4338pubmed: 28661493google scholar: lookup
  63. Li GZ. Database searching and accounting of multiplexed precursor and product ion spectra from the data independent analysis of simple and complex peptide mixtures.. Proteomics 2009;9:1696–1719.
    doi: 10.1002/pmic.200800564pubmed: 19294629google scholar: lookup
  64. Silva JC, Gorenstein MV, Li GZ, Vissers JP, Geromanos SJ. Absolute quantification of proteins by LCMSE: a virtue of parallel MS acquisition.. Mol Cell Proteomics 2006;5:144–156.
    doi: 10.1074/mcp.M500230-MCP200pubmed: 16219938google scholar: lookup
  65. Ritchie ME. limma powers differential expression analyses for RNA-sequencing and microarray studies.. Nucleic acids research 2015;43:e47.
    doi: 10.1093/nar/gkv007pmc: PMC4402510pubmed: 25605792google scholar: lookup
  66. Mi H, Muruganujan A, Casagrande JT, Thomas PD. Large-scale gene function analysis with the PANTHER classification system.. Nature protocols 2013;8:1551–1566.
    doi: 10.1038/nprot.2013.092pmc: PMC6519453pubmed: 23868073google scholar: lookup
  67. Huber W. Orchestrating high-throughput genomic analysis with Bioconductor.. Nature methods 2015;12:115–121.
    doi: 10.1038/nmeth.3252pmc: PMC4509590pubmed: 25633503google scholar: lookup
  68. Luo W, Friedman MS, Shedden K, Hankenson KD, Woolf PJ. GAGE: generally applicable gene set enrichment for pathway analysis.. BMC bioinformatics 2009;10:161.
    doi: 10.1186/1471-2105-10-161pmc: PMC2696452pubmed: 19473525google scholar: lookup