Skip to main content

Advertisement

SpringerLink
Log in

Elevated concentration of TNF-α induces trophoblast differentiation in mouse blastocyst outgrowths

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Elevated expression of tumour necrosis factor-α (TNF-α) is associated with adverse pregnancy outcome. This study has examined the expression of TNF-α and its receptors (TNF-Rs) by mouse blastocysts and blastocyst outgrowths from day 4 to 9.5 of pregnancy and investigated the effects of elevated TNF-α on the inner cell mass (ICM) and trophoblast cells of blastocyst outgrowths. RT-PCR demonstrated TNF-α mRNA expression from day 7.5 to 9.5, TNF-R1 from day 6.5 to 9.5 and TNF-R2 from day 5.5 to 7.5 of pregnancy, and in situ hybridisation revealed the trophoblast giant cells (TGCs) of the early placenta as the site of TNF-α expression. Day 4 blastocysts were cultured in a physiologically high concentration of TNF-α (100 ng/ml) for 72 h to the outgrowth stage and then compared to blastocysts cultured in media alone. TNF-α-treated blastocyst outgrowths exhibited a significant reduction in ICM cells (mean ± SD 23.90±10.42 vs 9.37±7.45, t-test, P<0.0001) with no significant change in the numbers of trophoblast cells (19.97±8.14 vs 21.73±7.79, t-test, P=0.39). Within the trophoblast cell population, the TNF-α-treated outgrowths exhibited a significant increase in multinucleated cells (14.10±5.53 vs 6.37±5.80, t-test, P<0.0001) and a corresponding significant decrease in mononucleated cells (5.87±3.60 vs 15.37±5.87, t-test, P<0.0001). In summary, this study describes the expression of TNF-α and its receptors during the peri-implantation period in the mouse. It also reports that elevated TNF-α restricts ICM proliferation in the blastocyst and changes the ratio of mononucleated to multinucleated trophoblast cells. These findings suggest a mechanism by which increased expression of TNF-α during trophoblast differentiation may be detrimental to pregnancy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2A, B.
Fig. 3A–C.

Similar content being viewed by others

References

  • Abrahamsohn PA, Zorn TM (1993) Implantation and decidualization in rodents. J Exp Zool 266:603–628

    PubMed  Google Scholar 

  • Argilés JM, Carbo N, Lopez-Soriano FJ (1997) TNF and pregnancy: the paradigm of a complex interaction. Cytokine Growth Factor Rev 8:181–188

    Article  PubMed  Google Scholar 

  • Bazzoni F, Beutler B (1996) The tumor necrosis factor ligand and receptor families. N Engl J Med 334:1717–1725

    CAS  PubMed  Google Scholar 

  • Chen G, Goeddel DV (2002) TNF-R1 signaling: a beautiful pathway. Science 296:1634–1635

    Article  CAS  PubMed  Google Scholar 

  • Cross JC (2000) Genetic insights into trophoblast differentiation and placental morphogenesis. Semin Cell Dev Biol 2:105–113

    Article  Google Scholar 

  • De M, Sanford TR, Wood GW (1993) Expression of interleukin 1, interleukin 6 and tumour necrosis factor alpha in mouse uterus during the peri-implantation period of pregnancy. J Reprod Fertil 97:83–89

    CAS  PubMed  Google Scholar 

  • Duclos AJ, Haddad EK, Baines MG (1995) Presence of activated macrophages in a murine model of early embryo loss. Am J Reprod Immunol 33:354–366

    CAS  PubMed  Google Scholar 

  • Erickson SL, De Sauvage FJ, Kirky K, Carver-Moore K, Pitts-Meek S, Gillet N, Sheehan KC, Schreiber RD, Goedel DV, Moore MW (1994) Decreased sensitivity to tumor necrosis factor but normal T-cell development in TNF receptor-2 deficient mice. Nature 372:560–563

    Google Scholar 

  • Gendron RL, Nestel FP, Lapp WS, Baines MG (1990) Lipopolysaccharide-induced fetal resorption in mice is associated with the intrauterine production of tumour necrosis factor-alpha. J Reprod Fertil 90:395–402

    CAS  PubMed  Google Scholar 

  • Gorivodsky M, Zemlyak I, Orenstein H, Savion S, Fein A, Torchinsky A, Toder V (1998) TNF-alpha messenger RNA and protein expression in the uteroplacental unit of mice with pregnancy loss. J Immunol 160:4280–4288

    CAS  PubMed  Google Scholar 

  • Hampson J, McLaughlin PJ, Johnson PM (1993) Low-affinity receptors for tumour necrosis factor-alpha, interferon-gamma and granulocyte-macrophage colony-stimulating factor are expressed on human placental syncytiotrophoblast. Immunology 79:485–490

    CAS  PubMed  Google Scholar 

  • Hogan B, Constantini F, Lacy E (1986) Manipulating the mouse embryo. In: A laboratory manual. Cold Spring Harbour Laboratory, New York

  • Hunt JS, Chen H, Hu X, Pollard JW (1993) Normal distribution of tumor necrosis factor-α messenger ribonucleic acid and protein in the uteri, placentas, and embryos of osteopetrotic (op/op) mice lacking colony stimulating factor-1. Biol Reprod 49:441–452

    CAS  PubMed  Google Scholar 

  • Kilpatrick DC (1999) Influence of human leukocyte antigen and tumour necrosis factor genes on the development of pre-eclampsia. Hum Reprod Update 5:94–102

    CAS  PubMed  Google Scholar 

  • Knofler M, Mosl B, Bauer S, Griesinger G, Husslein P (2000) TNF-alpha/TNFRI in primary and immortalized first trimester cytotrophoblasts. Placenta 21:525–535

    Article  CAS  PubMed  Google Scholar 

  • Kohchi C, Mizuno D, Soma G (1991) Expression of TNF-alpha and TNF-beta transcripts in murine embryonal carcinoma cells and trophoblast cells. Nucleic Acids Symp Ser 25:119–120

    CAS  PubMed  Google Scholar 

  • Kurzawa R, Glabowski W, Wenda-Rozewicka L (2001) Evaluation of mouse preimplantation embryos cultured in media enriched with insulin-like growth factors I and II, epidermal growth factor and tumor necrosis factor alpha. Folia Histochem Cytobiol 39:245–251

    CAS  PubMed  Google Scholar 

  • Lachapelle MH, Miron P, Hemmings R, Falcone T, Granger L, Bourque J, Langlais J (1993) Embryonic resistance to tumor necrosis factor-α mediated cytotoxicity: novel mechanism underlying maternal immunological tolerance to the fetal allograft. Hum Reprod 7:1032–1038

    Google Scholar 

  • Lea RG, Tulppala M, Critchley HO (1997) Deficient syncytiotrophoblast tumour necrosis factor-alpha characterises failing first trimester pregnancies in a subgroup of recurrent miscarriage patients. Hum Reprod 12:1313–1320

    CAS  PubMed  Google Scholar 

  • Lea RG, McIntyre S, Baird JD, Clark DA (1998) Tumor necrosis factor-alpha mRNA-positive cells in spontaneous resorption in rodents. Am J Reprod Immunol 30:50–57

    Google Scholar 

  • Leazer TM, Barbee B, Ebron-McCoy M, Henry-Sam GA, Rogers JM (2002) Role of the maternal acute phase response and tumor necrosis factor alpha in the developmental toxicity of lipopolysaccharide in the CD-1 mouse. Reprod Toxicol 16:173–179

    Article  CAS  PubMed  Google Scholar 

  • Old LJ (1985) Tumor necrosis factor (TNF). Science 230:630–632

    CAS  PubMed  Google Scholar 

  • Pampfer S, Wuu YD, Vanderheyden I, De Hertogh R (1994) Expression of tumor necrosis factor-alpha (TNF alpha) receptors and selective effect of TNF alpha on the inner cell mass in mouse blastocysts. Endocrinology 134:206–221

    CAS  PubMed  Google Scholar 

  • Pfeffer K, Matsuyama T, Kundig TM, Wakeham A, Kishihara K, Shahinian A, Wiegmann K, Ohashi PS, Kronke M, Mak TW (1993) Mice deficient for the 55 kd tumor necrosis factor receptor are resistant to endotoxic shock, yet succumb to L. monocytogenes infection. Cell 73:457–467

    CAS  PubMed  Google Scholar 

  • Rosenfeld ME, Prichard L, Shiojiri N, Fausto N (2000) Prevention of hepatic apoptosis and embryonic lethality in RelA/TNFR-1 double knockout mice. Am J Pathol 156:997–1007

    CAS  PubMed  Google Scholar 

  • Ryffel B, Eugster H, Haas C, Le Hir M (1998) Failure to induce anti-glomerular basement membrane glomerulonephritis in TNF alpha/beta deficient mice. Int J Exp Pathol 796:453–460

    Article  Google Scholar 

  • Sanford TR, De M, Wood GW (1992) Expression of colony-stimulating factors and inflammatory cytokines in the uterus of CD1 mice during days 1 to 3 of pregnancy. J Reprod Fertil 94:213–220

    CAS  PubMed  Google Scholar 

  • Shakhov AN, Nedospasov SA (2001) Expression profiling in knockout mice: lymphotoxin versus tumor necrosis factor in the maintenance of splenic microarchitecture. Cytokine Growth Factor Rev 12:107–119

    Article  CAS  PubMed  Google Scholar 

  • Tangri S, Raghupathy R (1993) Expression of cytokines in placentas of mice undergoing immunologically mediated spontaneous fetal resorptions. Biol Reprod 49:850–856

    CAS  PubMed  Google Scholar 

  • Telford NA, Hogan A, Franz CR, Schulz GA (1990) Expression of genes for insulin and insulin-like growth factors and receptors in early postimplantation mouse embryo and embryonal carcinoma cells. Mol Reprod Dev 26:81–92

    Google Scholar 

  • Vince GS, Starkey PM, Austgulen R, Kwiatkowski D, Redman CWG (1995) Interleukin-6, tumour necrosis factor and soluble tumour necrosis factor receptors in women with pre-eclampsia. Br J Obstet Gynaecol 102:20–25

    CAS  PubMed  Google Scholar 

  • Wuu Y-D, Pampfer S, Becquet P, Vanderheyden I, Lee K-H, De Hertogh R (1999) Tumor necrosis factor α decreases the viability of mouse blastocysts in vitro and in vivo. Biol Reprod 60:479–483

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We wish to thank Dr. Neil Richardson, Ms. Jane Easson and Mr. Joseph Kan (Queensland University of Technology) for their guidance in tissue preparation, embedding and sectioning. We also thank Associate Professor Chris O'Neill, Dr. Aviv Cahana and Dr. Margot Day (University of Sydney) for their assistance with trophoblast staining and/or manuscript advice.

Author information

Authors and Affiliations

  1. Cluster for Molecular Biotechnology, School of Life Sciences, Queensland University of Technology, 2 George St., 4001, Brisbane, Queensland, Australia

    Eliza J. Whiteside, Kerry J. Boucaut, Alison Teh, Juanita Garcia-Aragon, Mark B. Harvey & Adrian C. Herington

Authors
  1. Eliza J. Whiteside
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kerry J. Boucaut
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alison Teh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Juanita Garcia-Aragon
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mark B. Harvey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Adrian C. Herington
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Eliza J. Whiteside.

Additional information

This work was supported by the National Health and Medical Research Council of Australia

Rights and permissions

Reprints and permissions

About this article

Cite this article

Whiteside, E.J., Boucaut, K.J., Teh, A. et al. Elevated concentration of TNF-α induces trophoblast differentiation in mouse blastocyst outgrowths. Cell Tissue Res 314, 275–280 (2003). https://doi.org/10.1007/s00441-003-0791-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-003-0791-4

Keywords

Search

Navigation