Archives of Virology

, Volume 127, Issue 1–4, pp 37–48

Susceptibility of mouse embryo to murine cytomegalovirus infection in early and mid-gestation stages

  • A. Kashiwai
  • N. Kawamura
  • C. Kadota
  • Yoshihiro Tsutsui
Original Papers


The susceptibility of mouse embryonic cells to murine cytomegalovirus (MCMV) infection was studied by injecting the virus in the early and mid-gestation stages. For the early stage, blastocysts from BDF1 mice were injected with MCMV or minimal essential medium (MEM) by micromanipulator and returned to the uteri of pseudopregnant ICR mice. On day 11 of gestation, the embryos were examined immunohistochemically, using antibody specific to the early antigen of MCMV, and the placentae were examined by plaque assay. No infection was detected by either method. Furthermore, no infection was detected in MCMV-infected blastocysts that were cultured and examined for infection by immunofluorescence. For mid-gestation embryos, the conceptus was injected with MCMV on day 8.5 of gestation and was subjected to immunohistochemical analysis from day 10.5 to 12.5 of gestation. Viral antigen-positive cells were first observed in the placentae, then antigen-positive cells appeared among the blood cells, endothelial and mesodermal cells of the embryos. On day 12.5 of gestation, clusters of viral antigen-positive cells were sometimes observed in the hearts and livers. Although the incidence was lower, viral antigen-positive cells were also observed in the neuroectoderm and the eyes. These results suggest that MCMV does not infect early embryos and that infection first occurs in the placenta of postimplantation embryos, whence it extends through the blood cells to the endothelial and mesodermal cells of different embryonic regions, eventually extending to the neuroectoderm.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bale JF (1984) Human cytomegalovirus infection and disorders of the nervous system. Arch Neurol 41: 310–320Google Scholar
  2. 2.
    Baskar JF, Stanat SC, Sulik KK, Huang E-S (1983) Murine cytomegalovirus-induced congenital defects and fetal maldevelopment. J Infect Dis 148: 836–843Google Scholar
  3. 3.
    Baskar JF, Peacock J, Sulik KK, Huang E-S (1987) Early-stage development abnormalities induced by murine cytomegalovirus. J Infect Dis 155: 661–666Google Scholar
  4. 4.
    Bia FJ, Griffith BP, Fong CKY, Hsiug GD (1983) Cytomegalovirus infections in the guinea pig: experimental models for human disease. Rev Infect Dis 5: 177–195Google Scholar
  5. 5.
    Dankner WM, McCutchan JA, Richman DD, Hirata K, Spector SA (1990) Localization of human cytomegalovirus in peripheral blood leukocytes by in situ hybridization. J Infect Dis 161: 31–36Google Scholar
  6. 6.
    Ebihara K, Minamishima Y (1984) Protective effect of biological response modifiers on murine cytomegalovirus infection. J Virol 51: 117–122Google Scholar
  7. 7.
    Frohman MA, Boyle M, Martin GR (1990) Isolation of the mouse Hox-2.9 gene; analysis of embryonic expression suggests that positional information along the anterior-posterior axis is specified by mesoderm. Development 119: 589–607Google Scholar
  8. 8.
    Gonczol E, Andrews PW, Plotokin SA (1984) Cytomegalovirus replicates in differentiated but not in undifferentiated human embryonal carcinoma cells. Science 224: 159–161Google Scholar
  9. 9.
    Gonczol E, Andrews PW, Plotokin SA (1985) Cytomegalovirus infection of human teratocarcinoma cells in culture. J Gen Virol 66: 509–515Google Scholar
  10. 10.
    Griffith BP, McCormick SR, Fong CKY, Lavallee JT, Lucia HL, Goff E (1985) The placenta as a site of cytomegalovirus infection in guinea pigs. J Virol 55: 402–409Google Scholar
  11. 11.
    Griffith BP, McCormick SR, Booss J, Hsiung GD (1986) Inbred guinea pig model of intrauterine infection with cytomegalovirus. Am J Pathol 122: 112–119Google Scholar
  12. 12.
    Griffith BP, Chen M, Isom HC (1990) Role of primary and secondary maternal viremia in transplacental guinea pig cytomegalovirus transfer. J Virol 64: 1991–1997Google Scholar
  13. 13.
    Jaenisch R (1985) Mammalian neural crest cells participate in normal embryonic development on microinjection into post-implantation mouse embryos. Nature 318: 181–183Google Scholar
  14. 14.
    Jaenisch R, Fan H, Croker B (1975) Infection of preimplantation mouse embryos and of newborn mice with leukemia virus: tissue distribution of viral DNA and RNA and leukemogenesis in the adult animal. Proc Natl Acad Sci USA 72: 4008–4012Google Scholar
  15. 15.
    London WT, Martzinez AL, Houff SA, Wallaen WC, Curfman BL, Traub RG, Sever JL (1986) Experimental congenital disease with simian cytomegalovirus in rhesus monkeys. Teratology 33: 323–331Google Scholar
  16. 16.
    Lussier G (1975) Murine cytomegalovirus-induced cerebral calcification. Am J Pathol 80: 555–558Google Scholar
  17. 17.
    Martin GR (1980) Teratocarcinoma and mammalian embryogenesis. Science 209: 768–776Google Scholar
  18. 18.
    Kadota C, Nagahama M, Tsutsui Y (1992) Relationship between HOX2 homeobox gene expression and the human cytomegalovirus immediate early genes. J Gen Virol 73: 975–981Google Scholar
  19. 19.
    Kapasi K, Rice GPA (1988) Cytomegalovirus infection of peripheral blood mononuclear cells: effects of interleukin-1 and -2 production and responsiveness. J Virol 62: 3603–3607Google Scholar
  20. 20.
    Nagahama M, Kashiwai A, Kadota C, Tsutsui Y (1991) Viral DNA detected by in situ hybridization in the developing mouse brain infected with murine cytomegalovirus. Acta Pathol Jpn 41: 661–667Google Scholar
  21. 21.
    Naruse I, Tsutsui Y (1989) Brain abnormalities induced by murine cytomegalovirus injected into the cerebral ventricles of mouse embryos exo utero. Teratology 40: 181–189Google Scholar
  22. 22.
    Nelson JA, Reynolds-Kohler C, Smith BA (1987) Negative and positive regulation by a short segment in the 5′-flanking region of human cytomegalovirus major immediate-early gene. Mol Cell Biol 7: 4125–4129Google Scholar
  23. 23.
    Revello MG, Zavattoni M (1989) Correlation between immunofluorescence detection of human cytomegalovirus immediate early antigen in polymorphonuclear leukocytes and viremia. J Infect Dis 160: 159–160Google Scholar
  24. 24.
    Savatier P, Morgenstern J, Beddington RSP (1990) Permissiveness to murine leukemia virus expression during preimplantation and early postimplantation mouse development. Development 109: 655–665Google Scholar
  25. 25.
    Spindle A (1980) An improved culture medium for mouse blastocysts. In Vitro 16: 669–674Google Scholar
  26. 26.
    Stagno S, Pass RF, Dworsky ME, Britt WJ, Alford CA (1984) Congenital and perinatal cytomegalovirus infection: clinical characteristics and pathogenic factors. In: Plotokin SA, Michelson S, Pagano JS (ed) CMV: pathogenesis and prevention of human infection. AR Liss, New York, pp 65–85Google Scholar
  27. 27.
    Tsutsui Y, Naruse I (1987) Murine cytomegalovirus infection of cultured mouse embryos. Am J Pathol 127: 262–270Google Scholar
  28. 28.
    Tsutsui Y, Kashiwai A, Kawamura N, Nagahama N, Mizutani A, Naruse I (1989) Susceptibility of brain cells to murine cytomegalovirus infection in developing mouse brain. Acta Neuropathol 79: 262–270Google Scholar
  29. 29.
    Tsutsui Y, Kashiwai A, Kawamura N, Kadota C, Nagahama M (1991) Postnatal porencephaly induced in mouse by murine cytomegalovirus. Acta Neuropathol 82: 435–441Google Scholar
  30. 30.
    Waller TH (1971) The cytomegalovirus: Ubiquitous agents with protean clinical manifestation. N Engl J Med 285: 203–214Google Scholar
  31. 31.
    Wentworth BB, French L (1970) Plaque assay of cytomegalovirus strains of human origin. Proc Soc Exp Biol Med 135: 253–258Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • A. Kashiwai
    • 1
  • N. Kawamura
    • 1
  • C. Kadota
    • 1
  • Yoshihiro Tsutsui
    • 1
  1. 1.Institute for Developmental ResearchAichi Prefectural ColonyKasugai, AichiJapan

Personalised recommendations