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Expression of transforming growth factor alpha (TGF-α) gene in mouse embryonic development

  • Animal Experimentation
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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose: The expression of genes for TGF-α, epidermal growth factor (EGF), and the EGF receptor (EGFR) in mouse blastocysts was evaluated by the reverse transcription-polymerase chain reaction (RT-PCR). We evaluated the effects of TGF-α and EGF on the development of mouse embryo prior to implantation.

Results: The results revealed the presence of transcripts of TGF-α and EGFR. However, EGF mRNA was not observed in repeated experiments. None of these growth factors influenced the rate of development from the two-cell stage to the blastocyst stage when added to the culture medium. These effects were further examined on measuring the incorporation of tritiated thymidine and leucine, providing indices of the synthesis of DNA and protein, respectively. A concentration of only 0.1 ng/ml of TGF-α, which shares a cell surface receptor with EGF, stimulated the synthesis of both DNA and protein. EGF at a concentration of 10 ng/ml stimulated the synthesis of DNA and protein by blastocysts. To explore autocrine effects of TGF-α on the rate of blastocoel expansion, TGF-α antisense oligodeoxynucleotides was used to reduce expression of the TGF-α gene. TGF-α at a concentration of 0.1 ng/ml stimulates the rate of blastocoel expansion in early cavitating mouse blastocysts. In contrast, TGF-α antisense oligonucleotides significantly reduced the rate of expansion.

Conclusions: Our present observations suggest that TGF-α/EGF and the EGFR may be involved in regulating embryonic development. In particular, TGF-α may serve as an autocrine factor in the regulation of embryonic development.

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References

  1. 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;241:1823–1825

    Article  PubMed  CAS  Google Scholar 

  2. Telford NA, Hogan A, Franz CR, Schultz GA: Expression of genes for insulin and insulin-like growth factors and receptors in early postimplantation mouse embryos and embryonal carcinoma cells. Mol Reprod Dev 1990;27:81–92

    Article  PubMed  CAS  Google Scholar 

  3. Streffer C, van Beuningen D, Molls M, Zamboglou N, Schulz S: Kinetics of cell proliferation in the preimplanted mouse embryo in vivo and in vitro. Cell Tissue Kinet 1980;13:135–143

    PubMed  CAS  Google Scholar 

  4. Harada T, Tanikawa M, Iwabe T, Onohara Y, Mio Y, Terakawa N: Measurement of uptake and incorporation of nucleic acid precursors by preimplantation mouse embryos after development in vivo and in vitro. J Assist Reprod Genet 1992;9:551–556

    Article  PubMed  CAS  Google Scholar 

  5. Massip A, Van der Zwalman P, Puissant F, Camus M, Leroy R: Effects of in-vitro fertilization, culture, freezing and transfer on the ability of mouse embryos in vitro. J Reprod Fertil 1984;71:199–204

    Article  PubMed  CAS  Google Scholar 

  6. Paria BC, Dey SK: Preimplantation embryo development in vitro: Cooperative interactions among embryos and role of growth factors. Proc Natl Acad Sci USA 1990;87:4756–4760

    Article  PubMed  CAS  Google Scholar 

  7. Harvey MB, Kaye PL: Insulin stimulates protein synthesis in compacted mouse embryos. Endocrinology 1989;122:1182–1184

    Article  Google Scholar 

  8. Morishige K-I, Kurachi H, Amemiya K, Adachi H, 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;133:199–207

    Article  PubMed  CAS  Google Scholar 

  9. Chomczynsky P, Sacchi N: Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–159

    Google Scholar 

  10. Gray A, Dull T, Ullrich A: Nucleotide sequence of epidermal growth factor cDNA predicts a 128,000 molecular weight protein precursor. Nature 1983;303:722–726

    Article  PubMed  CAS  Google Scholar 

  11. Blasband AJ, Rogers KT, Chen X, Azizkhan JC, Lee DC: Characterization of the rat transforming growth factor alpha gene and identification of promoter sequences. Mol Cell Biol 1990;10:2111–2121

    PubMed  CAS  Google Scholar 

  12. Avivi A, Lax I, Ullrich A, Schlessinger J, Givol D, Morse B: Comparison of EGF receptor sequences as a guide to study of the ligand binding site. Oncogene 1991;6:673–676

    PubMed  CAS  Google Scholar 

  13. Tanikawa M: Effects of human serum supplementation on fertilization and development of mouse embryos. Asia-Ocean J Obstet Gynaecol 1994;20:209–215

    CAS  Google Scholar 

  14. Manejwala FM, Gragoe EJ, Schultz RA: Blastocoel expansion in the preimplantation mouse embryo: Role of extracellular sodium and chloride and possible apical routes of the entry. Dev Biol 1989;133:210–220

    Article  PubMed  CAS  Google Scholar 

  15. Dardik A, Schultz RM: Blastocoel expansion in the preimplantation mouse embryo: Stimulatory effect of TGF-α and EGF. Development 1991;113:919–930

    PubMed  CAS  Google Scholar 

  16. Burgess AW: Epidermal growth factor and transforming growth factor α. Br Med Bull 1989;45:401–424

    PubMed  CAS  Google Scholar 

  17. Werb Z: Expression of EGF and genes in early mammalian development. Mol Reprod Dev 1990;27:10–15

    Article  PubMed  CAS  Google Scholar 

  18. Kronmiller JE, Upholt WB, Kollar EJ: EGF antisense oligodeoxynucleotides block murine odontogenesis in vitro. Dev Biol 1991;147:485–488

    Article  PubMed  CAS  Google Scholar 

  19. Snead ML, Luo W, Oliver P, Nakamura M, Don-Wheeler G, Bessem C, Bell GI, Rall LB, Slavkin HC: Localization of epidermal growth factor in tooth and lung during embryonic mouse development. Dev Biol 1989;134:420–429

    Article  PubMed  CAS  Google Scholar 

  20. Warnburton D, Seth R, Shum L, Horcher PG, Hall FL, Werb Z, Slavkin HC: Epigenetic role of epidermal growth factor expression and signaling in embryonic mouse lung morphogenesis. Dev Biol 1992;149:123–133

    Article  Google Scholar 

  21. Wiley LM, Wu JX, Harari I, Adamson ED: Epidermal growth factor receptor and protein increase after the four-cell stage in murine development. Dev Biol 1992;149:247–260

    Article  PubMed  CAS  Google Scholar 

  22. Adamson ED, Deller MJ, Warshaw JB: Functional EGF receptors are present on mouse embryo tissues. Nature 1981;291:656–659

    Article  PubMed  CAS  Google Scholar 

  23. Adamson ED: EGF receptor activities in mammalian development. Mol Reprod Dev 1990;27:16–22

    Article  PubMed  CAS  Google Scholar 

  24. 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;131:947–957

    Article  PubMed  CAS  Google Scholar 

  25. 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 (TGFα) and EGF/TGFα receptors and125I-EGF binding sites in human fallopian tube. Biol Reprod 1994;50:1049–1058

    Article  PubMed  CAS  Google Scholar 

  26. Huet-Hudson YM, Chakraborty C, De SK, Suzuki Y, Andrews GK, Dey SK: Estrogen regulates synthesis of EGF in mouse uterine epitherial cells. Mol Endocrinol 1990;4:510–523

    PubMed  CAS  Google Scholar 

  27. 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;45:365–372

    Article  PubMed  CAS  Google Scholar 

  28. Caro CM, Trounson A, Kirby C: Effect of growth factors in culture medium on the rate of mouse embryo development and viability in vitro. J Vitro Fert Embryo Transfer 1987;4:265–268

    Article  CAS  Google Scholar 

  29. Wood SA, Kaye PL: Effects of epidermal growth factor on preimplantation mouse embryos. J Reprod Fertil 1989;85:575–582

    Article  PubMed  CAS  Google Scholar 

  30. Colver RM, Howe AM, McDonough PG, Boldt J: Influence of growth factors in defined culture medium on in vitro development of mouse embryos. Fertil Steril 1991;55:194–199

    PubMed  CAS  Google Scholar 

  31. Jung T, Fischer B, Beier HM: Quantitative aspects of protein synthesis in non-cultured and cultured rabbit blastocysts. Hum Reprod 1987;2:23–27

    PubMed  CAS  Google Scholar 

  32. Brucker C, Alexander NJ, Hodgen GD, Sandow BA: Transforming growth factor-alpha augments meiotic maturation of cumulus cell-enclosed mouse oocytes. Mol Reprod Dev 1991;28:94–98

    Article  PubMed  CAS  Google Scholar 

  33. Stern PH, Krieger NS, Nissenson RA, Williams RD, Winkle ME, Derynck R, Strewler GJ: Human transforming growth factor-α stimulates bone resorption in vitro. J Clin Invest 1985;76:2016–2019

    Article  PubMed  CAS  Google Scholar 

  34. Ibbotson KJ, Harrod J, Gowen M, D’Sonza S, Winkler ME, Derynck R, Mundy GR: Human recombinant transforming growth factor alpha stimulates bone resorption and inhibits formation in vitro. Proc Natl Acad Sci USA 1986;83:2228–2232

    Article  PubMed  CAS  Google Scholar 

  35. Ristimaki A: Transforming growth factor-α stimulates prostacyclin production by cellular human vascular endothelial cell more potently than epidermal growth factor. Biochem Biophys Res Commun 1989;160:1100–1105

    Article  PubMed  CAS  Google Scholar 

  36. Gan BS, Hollenberg MD, MacCannell KL, Lederis K, Winkler ME, Derynck R: Distinct vascular actions of epidermal growth factor-urogastrone and transforming growth factor-α. J Pharmacol Exp Ther 1987;242:331–337

    PubMed  CAS  Google Scholar 

  37. Schreiber AB, Winkler ME, Derynck R: Transforming growth factor-α: A more potent angiogenic mediator than epidermal growth factor. Science 1986;232:1250–1253

    Article  PubMed  CAS  Google Scholar 

  38. Winkler ME, O’Connor L, Winget M, Fendly B: Epidermal growth factor and transforming growth factor α bind differently to the epidermal growth factor receptor. Biochemistry 1989;28:6373–6378

    Article  PubMed  CAS  Google Scholar 

  39. Derynck R, Roberts AB, Winkler ME, Chen EY, Goeddel DV: Human transforming growth factor-α: Precursor structure and expression in E. coli. Cell 1984;38:287–297

    Article  PubMed  CAS  Google Scholar 

  40. Rizzino A: Early mouse embryos produce and release factors with transforming growth factor activity. In Vitro Cell Dev Biol 1985;21:531–536

    Article  PubMed  CAS  Google Scholar 

  41. 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;58:101–104

    PubMed  CAS  Google Scholar 

  42. Holt JT, Render RL, Nienhuis AW: An oligomer complementary to c-myc mRNA inhibits proliferation of HL-60 promyelotic cells and induces differentiation. Mol Cell Biol 1988;8:963–973

    PubMed  CAS  Google Scholar 

  43. Becker D, Meier CB, Herlyn M: Proliferation of human malignant melanomas is inhibited by antisense oligodeoxynucleotides targeted against basic fibloblast growth factor. EMBO J 1989;8:3685–3691

    PubMed  CAS  Google Scholar 

  44. Bevilacqua A, Erickson RP, Hieber V: Antisense RNA inhibits endogenous gene expression in mouse preimplantation embryos: Lack of double-stranded RNA “melting” activity. Proc Natl Acad Sci USA 1988;85:831–835

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Tottori University School of Medicine, 683, Yonago, Japan

    Tasuku Harada, Tomoyuki Fujikawa, Souichi Yoshida, Yoshimasa Onohara, Masahiro Tanikawa & Naoki Terakawa

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  1. Tasuku Harada
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  2. Tomoyuki Fujikawa
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  3. Souichi Yoshida
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  4. Yoshimasa Onohara
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  5. Masahiro Tanikawa
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  6. Naoki Terakawa
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Harada, T., Fujikawa, T., Yoshida, S. et al. Expression of transforming growth factor alpha (TGF-α) gene in mouse embryonic development. J Assist Reprod Genet 14, 262–269 (1997). https://doi.org/10.1007/BF02765827

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  • DOI: https://doi.org/10.1007/BF02765827

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