Advertisement

Journal of Assisted Reproduction and Genetics

, Volume 22, Issue 1, pp 25–32 | Cite as

Preliminary Experience with Low Concentration of Granulocyte-Macrophage Colony-Stimulating Factor: A Potential Regulator in Preimplantation Mouse Embryo Development and Apoptosis

  • Barry BehrEmail author
  • Stephen Mooney
  • Yan Wen
  • Mary Lake Polan
  • Hongbo Wang
Animal Experimentation

Abstract

Purpose: To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development of preimplantation mouse embryos.

Methods: Mouse 2-cell embryos were collected and cultured in P-1 medium supplemented with GM-CSF at different concentrations. Using reverse transcription-polymerase chain reaction, expression Bcl-2 and Bax mRNA in blastocyst were evaluated in the GM-CSF group and control group. Apoptosis detection was performed using the in situ apoptosis detection kit in mouse blastocysts. The statistical significance of the data was analyzed using t-test and chi-square test.

Results: The development of blastocyst increased to 89% in the addition of GM-CSF (0.125 ng/mL), compared to controlled group (80%). The number of cells staining for apoptosis was lower in GM-CSF group than that in the control group. Bcl-2 expression was found to be upregulated in blastocysts in the GM-CSF supplemented group compared to the control group.

Conclusion: These results suggest that GM-CSF might be an important regulator in embryo development.

KEY WORDS:

Apoptosis blastocyst development GM-CSF in vivo blastocyst 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

REFERENCES

  1. 1.
    Hardy K: Apoptosis in the human embryo. Rev Reprod 1999;4:125–134Google Scholar
  2. 2.
    Hardy K: Cell death in the mammalian blastocyst. Mol Hum Reprod 1997;3:919–925Google Scholar
  3. 3.
    Jurisicova A, Latham KE, Casper RF, Casper RF, Varmuza SL: Expression and regulation of genes associated with cell death during murine preimplantation embryo development. Mol Reprod Dev 1998;51(3):243–253Google Scholar
  4. 4.
    Kane MT, Morgan PM, Coonan C: Peptide growth factors and preimplantation development. Hum Reprod Update 1997;3:137–157Google Scholar
  5. 5.
    Zhao Y, Chegini N: Human fallopian tube expresses granulocyte-macrophage colony stimulating factor (GM-CSF) and GM-CSF alpha and beta receptors and contain immunoreactive GM-CSF protein. J Clin Endocrinol Metab 1994 Aug;79(2):662–665Google Scholar
  6. 6.
    Lopez AF, Shannon MF, Hercus T, Nicola NA, Cambareri B, Dottore M, Layton MJ, Eglinton L, Vadas MA: Residue 21 of human granulocyte-macrophage colony-stimulating factor is critical for biological activity and for high but not low affinity binding. EMBO J 1992 March;11(3):909–916Google Scholar
  7. 7.
    Robertson SA, Sjoblom C, Jasper MJ, Norman RJ, Seamark RF: Granulocyte-macrophage colony-stimulating factor promotes glucose transport and blastomere viability in murine preimplantation embryos. Biol Reprod 2001 April;64(4):1206–1215Google Scholar
  8. 8.
    Wislez M, Fleury-Feith J, Rabbe N, Moreau J, Cesari D, Milleron B, Mayaud C, Antoine M, Soler P, Cadranel J: Tumor-derived granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor prolong the survival of neutrophils infiltrating bronchoalveolar subtype pulmonary adenocarcinoma. Am J Pathol 2001 Oct;159(4): 1423–1433Google Scholar
  9. 9.
    Flad HD, Grage-Griebenow E, Petersen F, Scheuerer B, Brandt E, Baran J, Pryjma J, Ernst M: The role of cytokines in monocyte apoptosis. Pathobiology 1999;67(5–6):291–293Google Scholar
  10. 10.
    Epling-Burnette PK, Zhong B, Bai F, Jiang K, Bailey RD, Garcia R, Jove R, Djeu JY, Loughran TP Jr, Wei S: Cooperative regulation of Mcl-1 by Janus kinase/stat and phosphatidylinositol 3-kinase contribute to granulocyte-macrophage colony-stimulating factor-delayed apoptosis in human neutrophils. J Immunol 2001 June 15;166(12):7486–7495Google Scholar
  11. 11.
    Warner CM, Cao W, Exley GE, McElhinny AS, Alikani M, Cohen J, Scott RT, Brenner CA: Genetic regulation of egg and embryo survival. Hum Reprod 1998 June;13(Suppl 3):178–190; discussion 191–196Google Scholar
  12. 12.
    Exley GE, Tang C, McElhinny AS, Warner CM: Expression of caspase and BCL-2 apoptotic family members in mouse preimplantation embryos. Biol Reprod 1999 July;61(1):231–239Google Scholar
  13. 13.
    Sjoblom C, Wikland M, Robertson SA: Granulocyte-macrophage colony-stimulating factor promotes human blastocyst development in vitro. Hum Reprod 1999 Dec;14(12): 3069–3076Google Scholar
  14. 14.
    Krussel J, Behr B, Hirchenhain J, Wen Y, Milki AA, Cupisti S, Bielfeld P, Polan ML: Expression of vascular endothelial growth factor mRNA in human preimplantation embryos derived from tripronuclear zygotes. Fertil Steril 2000 Dec;74(6):1220–1226Google Scholar
  15. 15.
    Wrenzycki C, Wells D, Herrmann D, Miller A, Oliver J, Tervit R, Niemann H: Nuclear transfer protocol affects messenger RNA expression patterns in cloned bovine blastocysts. Biol Reprod 2001;65(1):309–317Google Scholar
  16. 16.
    Chia CM, Winston RM, Handyside AH: EGF, TGF-alpha and EGFR expression in human preimplantation embryos. Development 1995;121:299–307Google Scholar
  17. 17.
    Zhang X, Kidder GM, Watson AJ, Schultz GA, Armstrong DT: Possible roles of insulin and insulin-like growth factors in rat preimplantation development: Investigation of gene expression by reverse transcription-polymerase chain reaction. J Reprod Fert 1994;100:375–380Google Scholar
  18. 18.
    Ruef C, Coleman DL: Granulocyte-macrophage colony-stimulating factor: Pleiotropic cytokine with potential clinical usefulness. Rev Infect Dis 1990 Jan–Feb;12(1):41–62Google Scholar
  19. 19.
    Robertson SA, Mayrhofer G, Seamark RF: Ovarian steroid hormones regulate granulocyte-macrophage colony-stimulating factor synthesis by uterine epithelial cells in the mouse. Biol Reprod 1996 Jan;54(1):183–196Google Scholar
  20. 20.
    Imakawa K, Helmer SD, Nephew KP, Meka CS, Christenson RK: A novel role for GM-CSF: Enhancement of pregnancy specific interferon production, ovine trophoblast protein-1. Endocrinology 1993 April;132(4):1869–1871Google Scholar
  21. 21.
    Imakawa K, Carlson KD, McGuire WJ, Christenson RK, Taylor A: Enhancement of ovine trophoblast interferon by granulocyte macrophage-colony stimulating factor: Possible involvement of protein kinase C. J Mol Endocrinol 1997 Oct;19(2):121–130Google Scholar
  22. 22.
    Spanos S, Becker DL, Winston RM, Hardy K: Anti-apoptotic action of insulin-like growth factor-I during human preimplantation embryo development. Biol Reprod 2000 Nov;63(5):1413–1420Google Scholar
  23. 23.
    Wuu YD, Pampfer S, Becquet P, Vanderheyden I, Lee KH, De Hertogh R: Tumor necrosis factor alpha decreases the viability of mouse blastocysts in vitro and in vivo. Biol Reprod 1999 Feb;60(2):479–483Google Scholar
  24. 24.
    Lotem J, Sachs L: Hematopoietic cytokines inhibit apoptosis induced by transforming growth factor beta 1 and cancer chemotherapy compounds in myeloid leukemic cells. Blood 1992 Oct 1;80(7):1750–1757Google Scholar
  25. 25.
    Salveson GS, Dixit VM: Caspases: Intracellular signaling by proteolysis. Cell 1997;91:443–446Google Scholar
  26. 26.
    Adams JM, Cory S: The Bcl-2 protein family: Arbiters of cell survival. Science 1998;281:1322–1326CrossRefPubMedGoogle Scholar
  27. 27.
    Brison DR, Schultz RM: Increased incidence of apoptosis in transforming growth factor alpha-deficient mouse blastocysts. Biol Reprod 1998 July;59(1):136–144Google Scholar
  28. 28.
    Kamjoo M, Brison DR, Kimber SJ: Apoptosis in the preimplantation mouse embryo: Effect of strain difference and in vitro culture. Mol Reprod Dev 2002 Jan;61(1):67–77Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Barry Behr
    • 1
    Email author
  • Stephen Mooney
    • 1
  • Yan Wen
    • 1
  • Mary Lake Polan
    • 1
  • Hongbo Wang
    • 1
  1. 1.Stanford University School of MedicineStanford

Personalised recommendations