Analyze Diet
Biology of reproduction2014; 90(3); 61; doi: 10.1095/biolreprod.113.113928

RNA-seq transcriptome profiling of equine inner cell mass and trophectoderm.

Abstract: Formation of the inner cell mass (ICM) and trophectoderm (TE) marks the first differentiation event in mammalian development. These two cell types have completely divergent fates for the remainder of the developmental process. The molecular mechanisms that regulate ICM and TE formation are poorly characterized in horses. The objective of this study was to establish the transcriptome profiles of ICM and TE cells from horse blastocysts using RNA sequencing (RNA-seq). A total of 12 270 genes were found to be expressed in either lineage. Global analysis of the transcriptome profiles by unsupervised clustering indicated that ICM and TE samples presented different gene expression patterns. Statistical analysis indicated that 1662 genes were differentially expressed (adjusted P 2) between ICM and TE. Genes known to be specific to the ICM and TE were expressed primarily in their respective tissue. Transcript abundance for genes related to biological processes important for horse blastocyst formation and function is presented and discussed. Collectively, our data and analysis serve as a valuable resource for gene discovery and unraveling the fundamental mechanisms of early horse development.
Publication Date: 2014-03-20 PubMed ID: 24478389PubMed Central: PMC4435230DOI: 10.1095/biolreprod.113.113928Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • 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 research study used RNA sequencing to compare and analyze gene expressions in inner cell mass and trophectoderm cells, two early developmental cell types in horses, to help shed light on the biological processes important for horse embryo development.

Objective of the Study

  • The main objective of this research study was to understand the transcriptome profiles of the inner cell mass (ICM) and the trophectoderm (TE) cells in horse blastocysts. These are two critical cell types that form during the early stages of mammalian development and have completely different evolutionary paths throughout the rest of the development process.

Methodology

  • The researchers used a method called RNA sequencing (RNA-Seq) to accurately establish the transcriptome profiles of the ICM and TE cells.
  • A global analysis of the transcriptome profiles followed by unsupervised clustering was carried out to understand the expression patterns of genes in these cells.
  • Statistical analysis was performed to identify differentially expressed genes between ICM and TE.

Findings

  • Through this research, the team identified a total of 12,270 genes that were expressed in either the ICM or TE lineage.
  • The unsupervised clustering of the global transcriptome profiles revealed distinct gene expression patterns in the ICM and TE samples.
  • The statistical analysis indicated a significant number—1,662 genes—were differentially expressed between ICM and TE, with an adjusted P-value less than 0.05 and fold change greater than 2, two common statistics indicating significant differences.
  • Furthermore, it was found that genes specific to either the ICM and TE were mainly expressed in their respective tissue, further corroborating distinct roles and functions of these two cell types during development.

Conclusion

  • The overall results of the study and subsequent analysis provide much-needed understanding of gene expression in early horse development.
  • The research not only increases knowledge in this area but also serves as an extensive resource for gene discovery and understanding the core mechanisms that drive early development in horses.

Cite This Article

APA
Iqbal K, Chitwood JL, Meyers-Brown GA, Roser JF, Ross PJ. (2014). RNA-seq transcriptome profiling of equine inner cell mass and trophectoderm. Biol Reprod, 90(3), 61. https://doi.org/10.1095/biolreprod.113.113928

Publication

ISSN: 1529-7268
NlmUniqueID: 0207224
Country: United States
Language: English
Volume: 90
Issue: 3
Pages: 61
PII: 61

Researcher Affiliations

Iqbal, Khursheed
  • Department of Animal Science, University of California Davis, Davis, California.
Chitwood, James L
    Meyers-Brown, Geraldine A
      Roser, Janet F
        Ross, Pablo J

          MeSH Terms

          • Animals
          • Blastocyst / physiology
          • Blastocyst / ultrastructure
          • Blastocyst Inner Cell Mass / cytology
          • Blastocyst Inner Cell Mass / physiology
          • Chromosome Mapping
          • Embryonic Development / genetics
          • Embryonic Development / physiology
          • Female
          • Gene Amplification
          • Gene Expression Regulation, Developmental / genetics
          • Gene Expression Regulation, Developmental / physiology
          • Horses / physiology
          • Multigene Family
          • Pregnancy
          • RNA / genetics
          • Real-Time Polymerase Chain Reaction
          • Transcriptome / genetics
          • Transcriptome / physiology

          Grant Funding

          • R01 HD070044 / NICHD NIH HHS
          • R01HD070044 / NICHD NIH HHS

          References

          This article includes 32 references
          1. Giritharan G, Delle Piane L, Donjacour A, Esteban FJ, Horcajadas JA, Maltepe E, Rinaudo P. In vitro culture of mouse embryos reduces differential gene expression between inner cell mass and trophectoderm.. Reprod Sci 2012 Mar;19(3):243-52.
            pmc: PMC3343151pubmed: 22383776doi: 10.1177/1933719111428522google scholar: lookup
          2. Nagatomo H, Kagawa S, Kishi Y, Takuma T, Sada A, Yamanaka K, Abe Y, Wada Y, Takahashi M, Kono T, Kawahara M. Transcriptional wiring for establishing cell lineage specification at the blastocyst stage in cattle.. Biol Reprod 2013 Jun;88(6):158.
            pubmed: 23677984doi: 10.1095/biolreprod.113.108993google scholar: lookup
          3. Ozawa M, Sakatani M, Yao J, Shanker S, Yu F, Yamashita R, Wakabayashi S, Nakai K, Dobbs KB, Sudano MJ, Farmerie WG, Hansen PJ. Global gene expression of the inner cell mass and trophectoderm of the bovine blastocyst.. BMC Dev Biol 2012 Nov 6;12:33.
            pmc: PMC3514149pubmed: 23126590doi: 10.1186/1471-213x-12-33google scholar: lookup
          4. Betteridge KJ. Equine embryology: an inventory of unanswered questions.. Theriogenology 2007 Sep 1;68 Suppl 1:S9-21.
          5. Klein C, Troedsson MH. Transcriptional profiling of equine conceptuses reveals new aspects of embryo-maternal communication in the horse.. Biol Reprod 2011 May;84(5):872-85.
            pubmed: 21209420doi: 10.1095/biolreprod.110.088732google scholar: lookup
          6. Chitwood JL, Rincon G, Kaiser GG, Medrano JF, Ross PJ. RNA-seq analysis of single bovine blastocysts.. BMC Genomics 2013 May 25;14:350.
            pmc: PMC3668197pubmed: 23705625doi: 10.1186/1471-2164-14-350google scholar: lookup
          7. Meyers-Brown G, Bidstrup LA, Famula TR, Colgin M, Roser JF. Treatment with recombinant equine follicle stimulating hormone (reFSH) followed by recombinant equine luteinizing hormone (reLH) increases embryo recovery in superovulated mares.. Anim Reprod Sci 2011 Oct;128(1-4):52-9.
          8. Anders S, Huber W. Differential expression analysis for sequence count data.. Genome Biol 2010;11(10):R106.
            pmc: PMC3218662pubmed: 20979621doi: 10.1186/gb-2010-11-10-r106google scholar: lookup
          9. Huang DW, Sherman BT, Tan Q, Kir J, Liu D, Bryant D, Guo Y, Stephens R, Baseler MW, Lane HC, Lempicki RA. DAVID Bioinformatics Resources: expanded annotation database and novel algorithms to better extract biology from large gene lists.. Nucleic Acids Res 2007 Jul;35(Web Server issue):W169-75.
            pmc: PMC1933169pubmed: 17576678doi: 10.1093/nar/gkm415google scholar: lookup
          10. Huang da W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.. Nat Protoc 2009;4(1):44-57.
            pubmed: 19131956doi: 10.1038/nprot.2008.211google scholar: lookup
          11. Bermejo-Alvarez P, Rizos D, Rath D, Lonergan P, Gutierrez-Adan A. Sex determines the expression level of one third of the actively expressed genes in bovine blastocysts.. Proc Natl Acad Sci U S A 2010 Feb 23;107(8):3394-9.
            pmc: PMC2840439pubmed: 20133684doi: 10.1073/pnas.0913843107google scholar: lookup
          12. Zhang X, Wang L, Li X, Li K, Fang J, Yao Y. Ovarian stimulation retards postimplantation development and alters global gene expression profile of blastocysts in mouse.. Fertil Steril 2010 May 15;93(8):2770-3.
          13. Cockburn K, Rossant J. Making the blastocyst: lessons from the mouse.. J Clin Invest 2010 Apr;120(4):995-1003.
            pmc: PMC2846056pubmed: 20364097doi: 10.1172/jci41229google scholar: lookup
          14. Guo G, Huss M, Tong GQ, Wang C, Li Sun L, Clarke ND, Robson P. Resolution of cell fate decisions revealed by single-cell gene expression analysis from zygote to blastocyst.. Dev Cell 2010 Apr 20;18(4):675-85.
            pubmed: 20412781doi: 10.1016/j.devcel.2010.02.012google scholar: lookup
          15. Choi YH, Harding HD, Hartman DL, Obermiller AD, Kurosaka S, McLaughlin KJ, Hinrichs K. The uterine environment modulates trophectodermal POU5F1 levels in equine blastocysts.. Reproduction 2009 Sep;138(3):589-99.
            pubmed: 19525365doi: 10.1530/rep-08-0394google scholar: lookup
          16. Yamanaka Y, Lanner F, Rossant J. FGF signal-dependent segregation of primitive endoderm and epiblast in the mouse blastocyst.. Development 2010 Mar;137(5):715-24.
            pubmed: 20147376doi: 10.1242/dev.043471google scholar: lookup
          17. Keefer CL, Pant D, Blomberg L, Talbot NC. Challenges and prospects for the establishment of embryonic stem cell lines of domesticated ungulates.. Anim Reprod Sci 2007 Mar;98(1-2):147-68.
          18. Breton A, Sharma R, Diaz AC, Parham AG, Graham A, Neil C, Whitelaw CB, Milne E, Donadeu FX. Derivation and characterization of induced pluripotent stem cells from equine fibroblasts.. Stem Cells Dev 2013 Feb 15;22(4):611-21.
            pmc: PMC3564467pubmed: 22897112doi: 10.1089/scd.2012.0052google scholar: lookup
          19. Nagy K, Sung HK, Zhang P, Laflamme S, Vincent P, Agha-Mohammadi S, Woltjen K, Monetti C, Michael IP, Smith LC, Nagy A. Induced pluripotent stem cell lines derived from equine fibroblasts.. Stem Cell Rev Rep 2011 Sep;7(3):693-702.
            pmc: PMC3137777pubmed: 21347602doi: 10.1007/s12015-011-9239-5google scholar: lookup
          20. Enders AC, Schlafke S, Lantz KC, Liu IKM. Endoderm cells of the equine yolk sac from Day 7 until formation of the definitive yolk sac placenta.. Equine Vet J 1993;25:3–9.
          21. Fulka H, Mrazek M, Tepla O, Fulka J Jr. DNA methylation pattern in human zygotes and developing embryos.. Reproduction 2004 Dec;128(6):703-8.
            pubmed: 15579587doi: 10.1530/rep.1.00217google scholar: lookup
          22. Hong H, Takahashi K, Ichisaka T, Aoi T, Kanagawa O, Nakagawa M, Okita K, Yamanaka S. Suppression of induced pluripotent stem cell generation by the p53-p21 pathway.. Nature 2009 Aug 27;460(7259):1132-5.
            pmc: PMC2917235pubmed: 19668191doi: 10.1038/nature08235google scholar: lookup
          23. MacPhee DJ, Jones DH, Barr KJ, Betts DH, Watson AJ, Kidder GM. Differential involvement of Na(+),K(+)-ATPase isozymes in preimplantation development of the mouse.. Dev Biol 2000 Jun 15;222(2):486-98.
            pubmed: 10837135doi: 10.1006/dbio.2000.9708google scholar: lookup
          24. Flood PF, Betteridge KJ, Diocee MS. Transmission electron microscopy of horse embryos 3-16 days after ovulation.. J Reprod Fertil Suppl 1982;32:319-27.
            pubmed: 6962867
          25. Freeman DA, Butler JE, Weber JA, Geary RT, Woods GL. Co-culture of day-5 to day-7 equine embryos in medium with oviductal tissue.. Theriogenology 1991 Nov;36(5):815-22.
            pubmed: 16727050doi: 10.1016/0093-691x(91)90347-ggoogle scholar: lookup
          26. Oriol JG, Betteridge KJ, Clarke AJ, Sharom FJ. Mucin-like glycoproteins in the equine embryonic capsule.. Mol Reprod Dev 1993 Mar;34(3):255-65.
            pubmed: 8471247doi: 10.1002/mrd.1080340305google scholar: lookup
          27. Gillies LK, Waelchli RO, Ruddock WDJ, Betteridge KJ, LaMarre J. Patterns of MUC1 expression in the equine endometrium and trophoblast during early pregnancy.. Theriogenology 1999;51:225–225.
          28. Arar S, Chan KH, Quinn BA, Waelchli RO, Hayes MA, Betteridge KJ, Monteiro MA. Desialylation of core type 1 O-glycan in the equine embryonic capsule coincides with immobilization of the conceptus in the uterus.. Carbohydr Res 2007 Jun 11;342(8):1110-5.
            pubmed: 17335787doi: 10.1016/j.carres.2007.02.016google scholar: lookup
          29. Kopan R, Ilagan MX. The canonical Notch signaling pathway: unfolding the activation mechanism.. Cell 2009 Apr 17;137(2):216-33.
            pmc: PMC2827930pubmed: 19379690doi: 10.1016/j.cell.2009.03.045google scholar: lookup
          30. Brockhausen I, Schachter H, Stanley P. O-GalNAc glycans. Essentials of Glycobiology 2009;115 128.
          31. Klein C, Troedsson M. Equine pre-implantation conceptuses express neuraminidase 2--a potential mechanism for desialylation of the equine capsule.. Reprod Domest Anim 2012 Jun;47(3):449-54.
          32. Denker HW. Structural dynamics and function of early embryonic coats.. Cells Tissues Organs 2000;166(2):180-207.
            pubmed: 10729727doi: 10.1159/000016732google scholar: lookup

          Citations

          This article has been cited 28 times.
          1. Rocha RFB, Garcia AO, Otto PI, da Silva MVB, Martins MF, Machado MA, Panetto JCDC, Guimarães SEF. Runs of homozygosity and signatures of selection for number of oocytes and embryos in the Gir Indicine cattle. Mamm Genome 2023 Sep;34(3):482-496.
            doi: 10.1007/s00335-023-09989-wpubmed: 37000236google scholar: lookup
          2. Vegas AR, Podico G, Canisso IF, Bollwein H, Fröhlich T, Bauersachs S, Almiñana C. Dynamic regulation of the transcriptome and proteome of the equine embryo during maternal recognition of pregnancy. FASEB Bioadv 2022 Dec;4(12):775-797.
            doi: 10.1096/fba.2022-00063pubmed: 36479207google scholar: lookup
          3. Shen Y, Ren H, Davshilt T, Tian S, Wang X, Yi M, Ulaangerel T, Li B, Dugarjav M, Bou G. The transcriptome landscapes of allantochorion and vitelline-chorion in equine day 30 conceptus. Front Cell Dev Biol 2022;10:958205.
            doi: 10.3389/fcell.2022.958205pubmed: 35990610google scholar: lookup
          4. Derisoud E, Jouneau L, Dubois C, Archilla C, Jaszczyszyn Y, Legendre R, Daniel N, Peynot N, Dahirel M, Auclair-Ronzaud J, Wimel L, Duranthon V, Chavatte-Palmer P. Maternal age affects equine day 8 embryo gene expression both in trophoblast and inner cell mass. BMC Genomics 2022 Jun 15;23(1):443.
            doi: 10.1186/s12864-022-08593-7pubmed: 35705916google scholar: lookup
          5. Antczak DF, Allen WRT. Placentation in Equids. Adv Anat Embryol Cell Biol 2021;234:91-128.
            doi: 10.1007/978-3-030-77360-1_6pubmed: 34694479google scholar: lookup
          6. Camacho CA, Santos GO, Caballeros JE, Cazales N, Ramirez CJ, Vidigal PMP, Ramos HJO, Barros E, Mattos RC. Uterine infusion of conceptus fragments changes the protein profile from cyclic mares. Anim Reprod 2020 Nov 25;17(4):e20200552.
            doi: 10.1590/1984-3143-AR2020-0552pubmed: 33791032google scholar: lookup
          7. Hisey E, Ross PJ, Meyers SA. A Review of OCT4 Functions and Applications to Equine Embryos. J Equine Vet Sci 2021 Mar;98:103364.
            doi: 10.1016/j.jevs.2020.103364pubmed: 33663726google scholar: lookup
          8. Su J, Miao X, Archambault D, Mager J, Cui W. ZC3H4-a novel Cys-Cys-Cys-His-type zinc finger protein-is essential for early embryogenesis in mice†. Biol Reprod 2021 Feb 11;104(2):325-335.
            doi: 10.1093/biolre/ioaa215pubmed: 33246328google scholar: lookup
          9. Miao X, Sun T, Golan M, Mager J, Cui W. Loss of POLR1D results in embryonic lethality prior to blastocyst formation in mice. Mol Reprod Dev 2020 Nov;87(11):1152-1158.
            doi: 10.1002/mrd.23427pubmed: 33022126google scholar: lookup
          10. Todd ET, Thomson PC, Hamilton NA, Ang RA, Lindgren G, Viklund Å, Eriksson S, Mikko S, Strand E, Velie BD. A genome-wide scan for candidate lethal variants in Thoroughbred horses. Sci Rep 2020 Aug 4;10(1):13153.
            doi: 10.1038/s41598-020-68946-8pubmed: 32753654google scholar: lookup
          11. Pu Y, Zhang Y, Zhang T, Han J, Ma Y, Liu X. Identification of Novel lncRNAs Differentially Expressed in Placentas of Chinese Ningqiang Pony and Yili Horse Breeds. Animals (Basel) 2020 Jan 11;10(1).
            doi: 10.3390/ani10010119pubmed: 31940795google scholar: lookup
          12. Loux SC, Dini P, El-Sheikh Ali H, Kalbfleisch T, Ball BA. Characterization of the placental transcriptome through mid to late gestation in the mare. PLoS One 2019;14(11):e0224497.
            doi: 10.1371/journal.pone.0224497pubmed: 31725741google scholar: lookup
          13. Bogliotti YS, Chung N, Paulson EE, Chitwood J, Halstead M, Kern C, Schultz RM, Ross PJ. Transcript profiling of bovine embryos implicates specific transcription factors in the maternal-to-embryo transition. Biol Reprod 2020 Mar 13;102(3):671-679.
            doi: 10.1093/biolre/ioz209pubmed: 31711115google scholar: lookup
          14. Ntostis P, Kokkali G, Iles D, Huntriss J, Tzetis M, Picton H, Pantos K, Miller D. Can trophectoderm RNA analysis predict human blastocyst competency?. Syst Biol Reprod Med 2019 Aug;65(4):312-325.
            doi: 10.1080/19396368.2019.1625085pubmed: 31244343google scholar: lookup
          15. Ortiz-Rodriguez JM, Ortega-Ferrusola C, Gil MC, Martín-Cano FE, Gaitskell-Phillips G, Rodríguez-Martínez H, Hinrichs K, Álvarez-Barrientos A, Román Á, Peña FJ. Transcriptome analysis reveals that fertilization with cryopreserved sperm downregulates genes relevant for early embryo development in the horse. PLoS One 2019;14(6):e0213420.
            doi: 10.1371/journal.pone.0213420pubmed: 31237882google scholar: lookup
          16. Bonometti S, Menarim BC, Reinholt BM, Ealy AD, Johnson SE. Growth factor modulation of equine trophoblast mitosis and prostaglandin gene expression. J Anim Sci 2019 Feb 1;97(2):865-873.
            doi: 10.1093/jas/sky473pubmed: 30535412google scholar: lookup
          17. Read JE, Cabrera-Sharp V, Offord V, Mirczuk SM, Allen SP, Fowkes RC, de Mestre AM. Dynamic changes in gene expression and signalling during trophoblast development in the horse. Reproduction 2018 Oct 1;156(4):313–330.
            doi: 10.1530/REP-18-0270pubmed: 30306765google scholar: lookup
          18. Read JE, Cabrera-Sharp V, Offord V, Mirczuk SM, Allen SP, Fowkes RC, de Mestre AM. Dynamic changes in gene expression and signalling during trophoblast development in the horse. Reproduction 2018 Oct 1;156(4):313-330.
            doi: 10.1530/REP-18-0270pubmed: 29991567google scholar: lookup
          19. Ortega A, Tarazón E, Roselló-Lletí E, Gil-Cayuela C, Lago F, González-Juanatey JR, Cinca J, Jorge E, Martínez-Dolz L, Portolés M, Rivera M. Patients with Dilated Cardiomyopathy and Sustained Monomorphic Ventricular Tachycardia Show Up-Regulation of KCNN3 and KCNJ2 Genes and CACNG8-Linked Left Ventricular Dysfunction. PLoS One 2015;10(12):e0145518.
            doi: 10.1371/journal.pone.0145518pubmed: 26710323google scholar: lookup
          20. Stefaniuk M, Ropka-Molik K. RNA sequencing as a powerful tool in searching for genes influencing health and performance traits of horses. J Appl Genet 2016 May;57(2):199-206.
            doi: 10.1007/s13353-015-0320-7pubmed: 26446669google scholar: lookup
          21. Holl HM, Gao S, Fei Z, Andrews C, Brooks SA. Generation of a de novo transcriptome from equine lamellar tissue. BMC Genomics 2015 Oct 3;16:739.
            doi: 10.1186/s12864-015-1948-8pubmed: 26432030google scholar: lookup
          22. McQueen CM, Dindot SV, Foster MJ, Cohen ND. Genetic Susceptibility to Rhodococcus equi. J Vet Intern Med 2015 Nov-Dec;29(6):1648-59.
            doi: 10.1111/jvim.13616pubmed: 26340305google scholar: lookup
          23. Pacholewska A, Drögemüller M, Klukowska-Rötzler J, Lanz S, Hamza E, Dermitzakis ET, Marti E, Gerber V, Leeb T, Jagannathan V. The transcriptome of equine peripheral blood mononuclear cells. PLoS One 2015;10(3):e0122011.
            doi: 10.1371/journal.pone.0122011pubmed: 25790166google scholar: lookup
          24. Bakhtari A, Ross PJ. DPPA3 prevents cytosine hydroxymethylation of the maternal pronucleus and is required for normal development in bovine embryos. Epigenetics 2014 Sep;9(9):1271-9.
            doi: 10.4161/epi.32087pubmed: 25147917google scholar: lookup
          25. Moreton J, Malla S, Aboobaker AA, Tarlinton RE, Emes RD. Characterisation of the horse transcriptome from immunologically active tissues. PeerJ 2014;2:e382.
            doi: 10.7717/peerj.382pubmed: 24860704google scholar: lookup
          26. Filimonow K, Chołoniewska A, Chołoniewski J, Madeja ZE, Barłowska K, Grabarek J, Wenta-Muchalska E, Plusa B, Piliszek A. Stage-sensitive potential of isolated rabbit ICM to differentiate into extraembryonic lineages†. Biol Reprod 2025 Nov 14;113(5):1102-1120.
            doi: 10.1093/biolre/ioaf157pubmed: 40693734google scholar: lookup
          27. Pollard CL. Can Nicotinamide Adenine Dinucleotide (NAD(+)) and Sirtuins Be Harnessed to Improve Mare Fertility?. Animals (Basel) 2024 Jan 7;14(2).
            doi: 10.3390/ani14020193pubmed: 38254361google scholar: lookup
          28. Umair M, Scheeren VFDC, Beitsma MM, Colleoni S, Galli C, Lazzari G, de Ruijter-Villani M, Stout TAE, Claes A. In Vitro-Produced Equine Blastocysts Exhibit Greater Dispersal and Intermingling of Inner Cell Mass Cells than In Vivo Embryos. Int J Mol Sci 2023 Jun 1;24(11).
            doi: 10.3390/ijms24119619pubmed: 37298570google scholar: lookup