Autocrine Effects of Transforming Growth Factor-α on the Development of Preimplantation Mouse Embryos

  • Yoshimasa Onohara
  • Tasuku Harada
  • Masahiro Tanikawa
  • Tomio Iwabe
  • Hiroki Yoshioka
  • Fuminori Taniguchi
  • Masahiro Mitsunari
  • Toshiyuki Tsudo
  • Naoki Terakawa


Purpose: We wished to explore the role of transforming growth factor (TGF)-α in mouse embryonic development.

Methods: We examined the gene expression of TGF-α and epidermal growth factor receptor (EGFR) in mouse blastocysts by the reverse transcription–polymerase chain reaction and evaluated the effects of TGF-α on the development of preimplantation mouse embryos using TGF-α antisense oligodeoxynucleotide. Mouse teratocarcinoma F9 cells were also a subject of this study.

Results: Gene transcripts of TGF-α and EGFR were present in both blastocysts and F9 cells. TGF-α significantly stimulated the rate of blastocoel expansion in early-cavitating blastocysts and the proliferation of F9 cells. Northern blot analysis showed that TGF-α gene expression in F9 cells was markedly suppressed in the presence of TGF-α antisense oligodeoxynucleotide. TGF-α antisense oligonucleotide significantly reduced the rate of blastocoel expansion and the growth of F9 cells. The inhibitory effects of TGF-α antisense oligonucleotide on blastocysts and F9 cells were reversed by the addition of TGF-α.

Conclusions: The present observations suggest that TGF-α acts as an autocrine factor in the development of preimplantation mouse embryos.

transforming growth factor-α epidermal growth factor receptor preimplantation embryo teratocarcinoma F9 cell antisense oligodeoxynucleotide 


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  1. 1.
    Derynck R, Roberts AB, Winkler ME, Chen EY, Goeddel DV: Human transforming growth factor-α: Precursor structure and expression in E. coli. Cell 1984;38:287–297Google Scholar
  2. 2.
    Lee DC, Rose TM, Webb NR, Todaro GJ: Cloning and sequence analysis of a cDNA for rat transforming growth factor-α. Nature 1985;313:489–491Google Scholar
  3. 3.
    Marquardt H, Hunkapiller MW, Hood LE, Todaro G: Rat transforming growth factor type 1: Structure and relation to epidermal growth factor. Science 1984;223:1079–1082Google Scholar
  4. 4.
    Massague J: Epidermal growth factor-like transforming growth factor. II. Interaction with epidermal growth factor receptors in human placenta membranes and A431 cells. J Biol Chem 1983;258:13614–13620Google Scholar
  5. 5.
    Massague J: Transforming growth factor-α. A model for membrane-anchored growth factors. J Biol Chem 1990;265:21393–21396Google Scholar
  6. 6.
    Derynck R, Goeddel DV, Ullrich A, Gutterman JU, Williams RD, Bringman TS, Berger WH: Synthesis of messenger RNAs for transforming growth factor α and β and the epidermal growth factor receptor by human tumors. Cancer Res 1987;47:707–712Google Scholar
  7. 7.
    Adamson ED: Activities of growth factors in preimplantation embryos. J Cell Biochem 1993;53:280–287Google Scholar
  8. 8.
    Paria BC, Dey SK: Preimplantation embryo development in vitro: Cooperative interactions among embryos and role of growth factors. Proc Natl Acad Sci USA 1990;87:4756–4760Google Scholar
  9. 9.
    Dardik A, Schultz RM: Blastocoel expansion in the preimplantation mouse embryo: Stimulatory effect of TGF-α and EGF. Development 1991;113:919–930Google Scholar
  10. 10.
    Rappolee DA, Brenner CA, Schultz R, Mark D, Werb Z: Developmental expression of PDGF, TGF-α, and TGF-β genes in preimplantation mouse embryos. Science 1988;241:1823–1825Google Scholar
  11. 11.
    Wiley LM, Wu J-X, Harari I, Adamson ED: Epidermal growth factor receptor mRNA and protein increase after the four-cell preimplantation stage in murine development. Dev Biol 1992;149:247–260Google Scholar
  12. 12.
    Harada T, Fujikawa T, Yoshida S, Onohara Y, Tanikawa M, Terakawa N: Expression of transforming growth factor alpha (TGF-α) gene in mouse embryonic development. J Assist Reprod Genet 1997;14:262–269Google Scholar
  13. 13.
    Morishige K-I, Kurachi H, Amemiya K, Adachi H, Adachi K, Sakoyama Y, Miyake A, Tanizawa O: Menstrual stage-specific expression of epidermal growth factor and transforming growth factor-α in human oviduct epithelium and their role in early embryogenesis. Endocrinology 1993;133:199–207Google Scholar
  14. 14.
    Rappolee DA, Sturm KS, Behrendtsen O, Schultz GA, Pedersen RA, Werb Z: Insulin-like growth factor II acts through an endogenous growth pathway in early mouse embryos. Genes Dev 1992;6:939–952Google Scholar
  15. 15.
    Paria BC, Dey SK, Andrews GK: Antisense c-myc effects on preimplantation mouse embryo development. Proc Natl Acad Sci USA 1992;89:10051–10055Google Scholar
  16. 16.
    Brice, EC, Wu J-X, Muraro R, Adamson ED, Wiley LM: Modulation of mouse preimplantation development by epidermal growth factor receptor antibodies, antisense RNA, and deoxyoligonucleotides. Dev Genet 1993;14:174–184Google Scholar
  17. 17.
    Xu Y, Jin P, Warner CM: Modulation of preimplantation embryonic development by antisense oligonucleotides to major histocompatibility complex genes. Biol Reprod 1993;48:1042–1046Google Scholar
  18. 18.
    Pampfer S, Vanderheyden I, Vesela J, Hertogh RD: Neutralization of tumor necrosis factor α (TNF α) action on cell proliferation in rat blastocysts by antisense oligodeoxyribonucleotides directed against TNF α p60 receptor. Biol Reprod 1995;52:1316–1326Google Scholar
  19. 19.
    Vaughan TJ, Pascall JC, Brown KD: Nucleotide sequence and tissue distribution of mouse transforming growth factor-α. Biochim Biophys Acta 1992;1132:322–324Google Scholar
  20. 20.
    Chomczynsky P, Sacchi N: Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–159Google Scholar
  21. 21.
    Avivi A, Lax I, Ullrich A, Schlessinger J, Givol D, Morse B. Comparison of EGF receptor sequences as a guide to study the ligand binding site. Oncogene 1991;6:673–676Google Scholar
  22. 22.
    Tokunaga K, Taniguchi H, Yoda K, Shimizu M, Sakiyama S: Nucleotide sequence of a full-length cDNA for mouse cytoskeletal β-actin mRNA. Nucleic Acids Res 1986;14:2829Google Scholar
  23. 23.
    Martin GR: Teratocarcinoma and mammalian embryogenesis. Science 1980;290:768–776Google Scholar
  24. 24.
    Werb Z: Expression of EGF and TGF-α genes in early mammalian development. Mol Reprod Dev 1990;27:10–15Google Scholar
  25. 25.
    Adamson ED, Deller MJ, Warshaw JB: Functional EGF receptors are present on mouse embryo tissues. Nature 1981;291:656–659Google Scholar
  26. 26.
    Adamson ED: EGF receptor activities in mammalian development. Mol Reprod Dev 1990;27:16–22Google Scholar
  27. 27.
    Rizzino A: Early mouse embryos produce and release factors with transforming growth factor activity. In Vitro Cell Dev Biol 1985;21:531–536Google Scholar
  28. 28.
    Hemmings R, Langlais J, Falcone T, Granger L, Miron P, Guyda H: Human embryos produce transforming growth factor α activity and insulin-like growth factors II. Fertil Steril 1992;58:101–104Google Scholar
  29. 29.
    Burgess AW: Epidermal growth factor and transforming growth factor α. Br Med Bull 1989;45:401–424Google Scholar
  30. 30.
    Dardik A, Smith RM, Schultz RM: Colocalization of transforming growth factor-α and a functional epidermal growth factor receptor (EGFR) to the inner cell mass and preferential localization of the EGFR on the basolateral surface of the trophectoderm in the mouse blastocyst. Dev Biol 1992;154:396–409Google Scholar
  31. 31.
    Wood SA, Kaye PL: Effects of epidermal growth factor on preimplantation mouse embryos. J Reprod Fertil 1989;85:575–582Google Scholar
  32. 32.
    Biggers JD, Bell JE, Benos DJ: Mammalian blastocyst: Transport functions in a developing epithelium. Am J Physiol 1988;255:C419–C432Google Scholar
  33. 33.
    Manejwala FM, Cragoe EJ Jr, Schultz RM: Blastocoel expansion in the preimplantation mouse embryo: Role of extracellular sodium and chloride and possible apical routes of their entry. Dev Biol 1989;133:210–220Google Scholar
  34. 34.
    Stein CA, Cheng Y-C: Antisense oligonucleotides as therapeutic agent—Is the bullet really magical? Science 1993;261:1004–1012Google Scholar
  35. 35.
    Lei ZM, Rao ChV: Expression of epidermal growth factor (EGF) receptor and its ligands, EGF and transforming growth factor-α, in human fallopian tubes. Endocrinology 1992;131:947–957Google Scholar
  36. 36.
    Chegini N, Zhao Y, McLean FW: Expression of messenger ribonucleic acid and presence of immunoreactive proteins for epidermal growth factor (EGF), transforming growth factor alpha (TGF α) and EGF/TGF α receptors and 125I-EGF binding sites in human fallopian tube. Biol Reprod 1994;50:1049–1058Google Scholar
  37. 37.
    Huet-Hudson YM, Chakraborty C, De SK, Suzuki Y, Andrews GK, Dey SK: Estrogen regulates synthesis of EGF in mouse uterine epithelial cells. Mol Endocrinol 1990;4:510–523Google Scholar
  38. 38.
    Tamada H, Das SK, Andrews GK, Dey SK: Cell-type-specific expression of transforming growth factor-α in the mouse uterus during the peri-implantation period. Biol Reprod 1991;45:365–372Google Scholar

Copyright information

© Plenum Publishing Corporation 1998

Authors and Affiliations

  • Yoshimasa Onohara
    • 1
  • Tasuku Harada
    • 1
  • Masahiro Tanikawa
    • 1
  • Tomio Iwabe
    • 1
  • Hiroki Yoshioka
    • 1
  • Fuminori Taniguchi
    • 1
  • Masahiro Mitsunari
    • 1
  • Toshiyuki Tsudo
    • 1
  • Naoki Terakawa
    • 1
  1. 1.Department of Obstetrics and GynecologyTottori University School of MedicineYonagoJapan

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